Efficacy and mechanisms of cisplatin and sulforaphane nanoparticles in alleviating cisplatin resistance in non-small cell lung cancer

Introduction

Lung cancer has the highest incidence and mortality rates worldwide, and represents a major global health challenge (1). In 2020, lung cancer claimed more lives than any other cancers, accounting for 18% of cancer-related deaths worldwide (2). As a result, it has been prioritized as a health concern by the World Health Organization (2). In China, the dual burden of increasing incidence and mortality rates underscores the urgent need for effective interventions (3). The risk of lung cancer is exacerbated by pervasive factors such as smoking and air pollution (3). Non-small cell lung cancer (NSCLC) is the most common type of lung cancer, accounting for approximately 85% of all lung cancer diagnoses (4). Over 60% of NSCLC patients present with or develop metastasis; thus, NSCLC represents the most lethal form of the disease and poses a significant threat to patient survival (4).

Despite established multi-modal treatment regimens, the therapeutic options for NSCLC remain limited. Common chemotherapeutic drugs like cisplatin, also known as diamminedichloroplatinum (II) (DDP), cause severe systemic toxicity, which is particularly problematic for the large elderly patient population who display reduced tolerance (5). Additionally, the pervasive challenge of drug resistance severely limits the long-term efficacy of these drugs (6). Consequently, NSCLC often remains a refractory condition, and innovative strategies need to be developed to overcome the persistent challenges facing by current therapies (7).

It is in this context that nanomaterial synthesis has emerged as an important area of research. Nanotechnology-enabled targeted delivery systems could revolutionize NSCLC treatment by enhancing tumor-specific drug enrichment, improving targeting precision and biocompatibility, and thereby significantly mitigating the toxic effects on healthy tissues (8). Furthermore, optimized nanocarriers have the potential for synergistic combination with emerging modalities like immunotherapy, which could provide a novel approach for addressing drug resistance and debilitating side effects (9-12). Nanocarriers provide an advanced delivery system; however, potent, selective, and less toxic therapeutic agents are needed to be found to load into these systems. Consequently, attention has been directed toward bioactive compounds such as sulforaphane (SFN), which have shown promising anti-cancer effects, particularly against NSCLC, and could work synergistically with nanotechnology-based treatments.

SFN, a bioactive isothiocyanate derived from cruciferous vegetables, exhibits potent multi-target anti-cancer activities, and has shown significant efficacy against NSCLC (13-16). In NSCLC, SFN induces apoptosis and cell cycle arrest, sensitizing tumor cells to ferroptosis via glutathione peroxidase 4 (GPX4) inhibition, glutathione depletion, and the dysregulation of iron metabolism/lipid peroxidation (17-19), and remodels the immunosuppressive tumor microenvironment (TME) by inhibiting angiogenesis and immunosuppressive cells (20). Given its pleiotropic effects, natural origin, and favorable safety profile, SFN is a promising adjuvant or core component for NSCLC therapy (21-24). However, its clinical application has been hampered by poor pharmacokinetics, including rapid metabolism, low systemic bioavailability, and insufficient tumor-specific accumulation. This limitation is particularly pertinent in overcoming chemoresistance, exemplified by the challenges of cisplatin (DDP), a first-line NSCLC agent with dose-limiting toxicity (e.g., nephrotoxicity and myelosuppression), and complex resistance mechanisms involving enhanced DNA repair, altered drug transport, and apoptotic evasion (25-31). Given its ability to modulate key resistance pathways, SFN could increase the efficacy of DDP (32); however, advanced strategies are needed to ensure the effective co-delivery of these agents, addressing their respective bioavailability and targeting deficits. Nanomaterial-based co-delivery systems present a viable solution, as such systems enable synchronized tumor targeting, protection, and the controlled release of SFN and DDP to maximize therapeutic synergy and overcome resistance (33-35).

Metal-organic frameworks (MOFs) constitute a class of crystalline porous materials with periodic networks formed through metal-ligand coordination, exhibiting record-high surface areas (>7,000 m²/g) and stimuli-responsive pore environments that enable targeted applications in oncology (36-39). Due to their pH/ATP dual-responsive drug release, exemplified by ZIF-90’s tumor-selective degradation (40), passive lung targeting via size exclusion (10–200 nm), and surface engineering (41), MOF-based nanocarriers may have unique advantages in the treatment of cancer, particularly NSCLC, a malignancy characterized by metastasis in >60% of initial diagnoses (4). However, their intrinsic limitations impede their clinical application. Notably, colloidal instability in physiological fluids induces premature payload leakage, while residual metal ions may trigger oxidative stress and inflammatory cascades (42). Together, DDP resistance, which is mediated by nucleotide excision repair upregulation and copper transporter CTR1 downregulation (27-29), dose-limiting nephrotoxicity, and the low oral bioavailability of SFN (<10%) undermine chemo-sensitization and ferroptosis-inducing functions in NSCLC therapy, reducing its efficacy (18).

To address these interconnected issues, this study pioneered a biomimetic ZIF-90 co-delivery system engineered for the synchronized pulmonary co-delivery of SFN and DDP. Using ZIF-90’s microenvironment-triggered disassembly in ATP-enriched acidic tumors (43-45), the system enables the spatiotemporal coordination of SFN-mediated glutathione depletion/GPX4 inactivation (19) and DDP-induced DNA crosslinking, thereby concurrently overcoming chemoresistance through ferroptosis sensitization while bypassing DNA repair mechanisms. Surface modification with dipalmitoyl phosphatidylcholine—mimicking alveolar surfactant composition—redirects biodistribution away from renal tissues toward lung neoplasms, achieving a more than five-fold increase in pulmonary accumulation compared to that of non-targeted systems (46). This integrated approach aims to address current NSCLC treatment issues by simultaneously eliminating drug resistance drivers, abolishing off-target toxicity, and establishing a transformative platform for phytochemical-chemotherapeutic synergy. We present this article in accordance with the ARRIVE and MDAR reporting checklists (available at https://tlcr.amegroups.com/article/view/10.21037/tlcr-2025-1123/rc).

Methods

Nanostructured sulforaphane and DDP particle (nSDDP) preparation and characterization

nSDDP was synthesized using a modified ZIF-90 method. Non-targeted nanoparticles were loaded with SFN and DDP, while epidermal growth factor receptor (EGFR)-targeted nanoparticles (nSDDPs) were functionalized via post-synthetic modification.

nSDDP synthesis

A mixture of 10 mg of DDP, 10 mg of SFN, 120 mg of imidazole-2-carbaldehyde, and 12.5 mg of polyethylene glycol was magnetically stirred for 1 min. The product was collected via centrifugation (14,000 g, 30 min ×3) and vacuum-dried at −55 ℃ for 12 h.

Targeted modification

nSDDPs (4 mg) were sonicated in 500 µL methanol, reacted with EGFR peptide solution (500 μL of methanol), and purified by centrifugation (10,500 g, 5 min).

Characterization

The samples were washed with methanol and then dried, after which, their morphology was analyzed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). nSDDPs were scanned from 4,000–400 cm⁻¹ by Fourier transform infrared spectroscopy (FT-IR). The samples were dispersed in water (30 min sonication) and their particle size and zeta potential were analyzed via Malvern Zetasizer (Malvern Panalytical Ltd., England). The ground samples were scanned from 5–40° (2θ) X-ray diffraction (XRD). The samples were monitored using dynamic light scattering (DLS) and TEM in aqueous dispersion to assess stability.

Drug release studies

To assess pH responsiveness, 10-mg samples were incubated in pH 5.5/6.8/7.4 buffers at 37 ℃, and the aliquots were analyzed by ultraviolet-visible (UV-Vis) spectroscopy. To assess ATP responsiveness, the samples were dispersed in 0–2 mM ATP buffers and analyzed similarly.

Loading efficiency

The samples were acid-digested (in 1 M hydrochloric acid), and drug concentrations were measured by UV-Vis spectroscopy. Encapsulation efficiency (EE) and loading capacity were calculated using standard formulas.

Labeled nanoparticles

Nile red was synthesized by replacing drugs with 10 mg of Nile Red during preparation, while IR-780 was prepared using 5 mg of IR-780 following identical procedures, and both were purified via centrifugation (14,000 g, 30 min ×3) and dried at −55 ℃.

Cell culture and experimental setup

The A549/DDP cells (CVCL_A549) were cultured in RPMI-1640 supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin at 37 ℃ in a 5% CO₂ incubator. For cell handling, the frozen cells were rapidly thawed in a 37 ℃ water bath, transferred to fresh medium, and centrifuged to remove the cryopreservation solution. The logarithmic-phase cells were cryopreserved in a solution containing 10% dimethyl sulfoxide (DMSO) and 90% FBS, while subculturing was performed by trypsin digestion at 80–90% confluency and subculturing was performed at a 1:3 or 1:4 ratio.

In vitro cytotoxicity and uptake assessment

To assess the cytotoxicity of nSDDP empty carriers on different cell lines, A549/DDP, GC, TM3, HEK293 and HepG2 cells were seeded at 5,000 cells/well in 96-well plates and then treated with nSDDP empty carriers at 1.25–160 µg/mL (three replicates per group). After incubation at 37 ℃ with 5% CO₂ for 24 or 48 h, Cell Counting Kit 8 (CCK-8) reagent was added, and absorbance at 450 nm was measured to calculate cell viability and generate dose-response curves.

To assess the uptake and localization of fluorescently labeled nSDDP in A549/DDP cells, the labeled nSDDP was added to A549/DDP cells in 24-well plates. After incubation, the cells were washed, stained with mitochondrial probes and 4',6-diamidino-2-phenylindole (DAPI), and imaged using confocal laser scanning microscope (CLSM) to analyze uptake efficiency, subcellular distribution (mitochondria and nucleus), as well as the effects of EGFR modification, ATP inhibition and endocytosis inhibition on uptake mechanisms.

In vivo biodistribution

All the animal experiments were conducted in compliance with institutional guidelines for the care and use of animals, and were approved by the Institutional Research Ethics Committee of Harbin Medical University Cancer Hospital (No. KY2022-71, 2022/11/3). Six-to-eight-week-old BALB/c mice were purchased from GemPhamatech Co., Ltd. (Nanjing, China). Mice with abnormal body weight or underlying diseases were excluded. The mice were housed in an animal facility at the Harbin Medical University Cancer Hospital under controlled conditions (22±2 ℃, 50–55% relative humidity, with a 12-h light/dark cycle), with free access to food and water. A total of 120 mice were randomly (SPSS Version 26.0) divided into 4 groups, namely the control group, nSDDP group, DDP group, and SFN group, with 30 mice in each group for different scientific research experiments. IR-780-labeled nSDDPs were intravenously injected into each mouse at 10 mg/kg. At 12 and 48 h post-injection, major organs were harvested, rinsed with saline, and imaged using a small-animal fluorescence imaging system to determine the biodistribution patterns. Tissue samples were collected in aliquots of 100 mg, with several aliquots prepared, and then stored in a refrigerator. The study protocol was prepared before the study without registration.

In vivo safety evaluation

The mice were divided into experimental groups, which received nSDDPs (0, 12.5, 25, or 50 mg/kg via the tail vein), and a control group, which received saline. At 1, 7, 14, and 28 days post-administration, the mice were sacrificed, and their major organs were collected, fixed in 10% formalin, processed via dehydration, paraffin embedding, and sectioning (4 µm). Hematoxylin and eosin (H&E) staining was performed, and tissue sections were examined under an optical microscope to assess pathological changes and determine the biocompatibility and safety of the nSDDPs.

Cell-based efficacy and resistance assessment of nSDDPs

The A549/DDP cells were cultured in RPMI-1640 supplemented with 10% FBS and 1% penicillin/streptomycin at 37 ℃ in 5% CO₂.

Drug dosage screening

nSDDP (0–16 µM) was tested. After 24–72 h of incubation, cell viability was measured using CCK-8 assays to determine the optimal dosage [half-maximal inhibitory concentration (IC₅₀)] for subsequent experiments.

Functional assays

Cell migration was evaluated via scratch assays to measure wound closure after 24 h of treatment with DDP, SFN, or nSDDPs. The secretion levels of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9), vascular endothelial growth factor (VEGF), and transforming growth factor beta (TGF-β) were quantified from cell supernatants. Cell viability, proliferation (5-ethynyl-2'-deoxyuridine assay, EdU assay), lactate dehydrogenase (LDH) release, and apoptosis (via propidium iodide staining) were analyzed to assess the effects of the nSDDPs on the DDP-treated cells. Additionally, the intracellular levels of arachidonic acid (AA), iron ions, transferrin receptor 1 (TFR1), malondialdehyde (MDA), H₂O₂, and GPX4 enzyme activity were measured using commercial kits, with data analyzed by one-way analysis of variance (ANOVA) (a P value <0.05 was considered statistically significant).

In vivo efficacy and resistance evaluation

Tumor model and treatment

A549 cells (1×10⁷ cells/100 µL) were subcutaneously injected into the BALB/c nude mice. After tumor establishment, the mice were randomly allocated to four groups: control (saline), nSDDP (5 mg/kg), DDP (5 mg/kg), and SFN (10 mg/kg), treated via intraperitoneal injection for 28 days. Tumor volume (V = length × width²/2) and body weight were monitored weekly; tumor growth inhibition rates were calculated, and final tumor weights were compared.

Tissue analysis

At the end of the study, the tumors were fixed in 4% paraformaldehyde, dehydrated, embedded in paraffin, and sectioned (4 µm). H&E staining was used to evaluate the histological changes, while immunofluorescence was used to detect MMPs, E-cadherin, TGF-β, VEGF, and DAPI. GPX4 inhibition levels were measured using enzyme-linked assays. For the ultrastructural analysis, tumor mitochondria were examined by TEM after glutaraldehyde fixation and epoxy resin embedding. Western blot and reverse transcription quantitative polymerase chain reaction (RT-qPCR) were used to quantify the iron death-related proteins [e.g., GPX4 (ab125066), xCT (ab307601)] and messenger RNAs (mRNAs), respectively, to explore resistance mechanisms.

Statistical analysis

Data were presented as the mean ± standard deviation. GraphPad Prism 8.0 was used for graphing and one-way ANOVA was used for significance testing [^## indicate a significant difference (P<0.01) vs. the control group, while ^†† denote a significant difference (P<0.01) between nSDDP and DDP groups]. The experiment was independently replicated for 3 times (biological replicates) in the laboratory.

Results

Synthesis of the material

To ensure nanocarrier structural integrity during functionalization (which is essential for drug delivery efficiency), SEM imaging was used to confirm uniform porous morphologies across the MOF (121±5 nm), MOF-EGFR (120±7 nm), and nSDDPs (124±6 nm). The results showed preserved core architecture post-EGFR conjugation and drug loading (DL) (Figure 1A). The validation of EGFR ligand grafting (which is critical for active tumor targeting) via FT-IR spectroscopy revealed complete C=O peak disappearance at 1,720 cm⁻¹ in functionalized materials, establishing covalent modification for tumor-specific recognition (Figure 1B). The XRD analysis showed crystalline order preservation through identical diffraction peaks, ensuring predictable drug release kinetics (Figure 1C). Nanoparticle aggregation impedes tumor penetration but DLS confirms sustained colloidal stability, specifically that the hydrodynamic diameters of MOF and nSDDP remained stable at 110.3±1.2 and 110.8±1.5 nm respectively with a Polydispersity Index (PDI) <0.2 over 72 h, indicating the nanocarriers have enhanced anti-aggregation capacity to support prolonged circulation (Figure 1D,1E). Capitalizing on the elevated ATP in the NSCLC cells (3–10 mM), ATP-response tests triggered 92.4%±3.1% DDP release at 10 mM ATP (vs. 18.2%±2.7% in controls), establishing ATP-activated tumor-specific cytotoxicity (Figure 1F). To exploit acidic TMEs (pH 5.0–6.5) for selective release, pH-dependent assays showed <15% cumulative DDP release at pH 7.4 vs. >80% at pH 5.0 within 24 h, confirming TME-activated targeting minimizing off-target toxicity (Figure 1G). High performance liquid chromatography (HPLC) quantification demonstrated progressive payload optimization: nSDDP achieved superior DL (32.1%±0.8%) and EE (38.3%±1.2%) compared to MOF (23.0%±1.1% DL; 32.0%±0.9% EE) and MOF-EGFR (25.2%±0.7% DL; 34.1%±1.0% EE), which showed that the surface engineering had maximized the therapeutic payload (Figure 1H). As shown in Figure 1I,1J, the nanoparticles remained uniformly distributed without aggregation after 7, 14, 21, and 28 days. There were no significant changes in particle size or PDI, indicating good stability. Crucially, pH/ATP dual-responsiveness ensured >90% drug release exclusively in tumor cells, overcoming the two key limitations of conventional platinum therapy. Collectively, the nSDDPs integrate structural stability, stimuli-triggered intelligence, and synergistic targeting for precision NSCLC treatment.

Figure 1 Material characterization of the SDDP nanoparticles (nSDDPs). (A) SEM test results for nSDDPs; (B) detection results for the nSDDPs infrared spectrum analysis, the double asterisks (**) represent the specific absorption peak of nanoparticles in the detection results, namely the signal peak of the aldehyde group; (C) X-ray diffraction results for MOF-EGFR and nSDDPs; (D) nSDDPs particle size distribution test results; (E) zeta potential detection of nSDDP; (F) detection of nSDDPs release under different ATP conditions; (G) detection of nSDDPs release under different acidic conditions; (H) test results showing the drug loading and encapsulation efficiency of MOF, MOF-EGFR, and nSDDPs; (I) SEM images of nSDDPs at 7, 14, 21, and 28 days; (J) PDI coefficient and particle size changes of nSDDPs at 7, 14, 21 and 28 days. Data are presented as the mean ± standard deviation (n=3). DMEM, Dulbecco’s Modified Eagle Medium; EGFR, epidermal growth factor receptor; MOF, metal-organic framework; nSDDP, nanostructured sulforaphane and diamminedichloroplatinum (II) particle; PBS, phosphate-buffered saline; PDI, Polydispersity Index; SEM, scanning electron microscopy.

Safety assessment of the material

Antibody receptor-targeted nanocarriers exploit ligand-specific recognition to enhance therapeutic precision. To achieve selective drug accumulation in NSCLC while minimizing systemic toxicity—a critical advantage for overcoming the dose-limiting side effects of DDP—we engineered nSDDPs with EGFR-targeting functionality. The validation of the tumor-specific binding capability, essential for reducing off-target accumulation, was confirmed through competitive receptor blocking assays that showed substantially reduced cellular uptake in EGFR-saturated environments. A confocal microscopy analysis was used to verify the subcellular targeting precision, and showed that the nSDDPs exhibited significantly higher uptake in the A549/DDP cells compared to the non-targeted controls with significant fluorescence co-localization near mitochondria (Figure 2A). This spatial alignment with ATP-rich organelles directly leverages ATP-responsive release kinetics to maximize tumor-selective cytotoxicity.

Figure 2 Intracellular localization and uptake mechanism of nSDDPs. (A) Detection results of nSDDPs localization in the A549/DDP cells (scale bar =10 µm). The quantitative graph indicates that the intracellular fluorescence intensity gradually increases over time. The double asterisks (**) represent an extremely significant difference in fluorescence intensity compared with the initial state. (B) Detection results of the nSDDPs uptake mechanism of the A549/DDP cells (scale bar =10 µm); (C) detection of the effect of targeting molecules on the cellular uptake efficiency of the MOF nanoparticles (scale bar =10 µm); (D) detection of the effect of ATP on the release of the nSDDPs in the A549/DDP cells (scale bar =10 µm). Data are presented as the mean ± standard deviation (n=3). Mito-Tracker was used for mitochondrial localization staining, and Nile Red dye was employed for staining the nanomaterials. CPZ, chlorpromazine; DAPI, 4',6-diamidino-2-phenylindole; DDP, diamminedichloroplatinum (II); MOF, metal-organic framework; nSDDP, nanostructured sulforaphane and diamminedichloroplatinum (II) particle.

Immunofluorescence staining assay was used to quantify the internalization dynamics and showed pronounced time-/dose-dependent uptake. Specifically, nSDDP accumulation increased substantially over time and concentration gradients, demonstrating programmable delivery control critical for dosage optimization. The elucidation of cellular entry mechanisms, fundamental for delivery system refinement, employed pharmacological inhibition. Nystatin-mediated lipid raft disruption significantly reduced nSDDP uptake, while the dynasore inhibition of dynamin GTPase activity significantly decreased internalization, but the reduction in the non-targeted MOFs was minimal (Figure 2B). This confirmed that the EGFR-modified nSDDPs primarily used lipid raft-mediated and dynamin-dependent endocytosis, with the targeting specificity substantially enhancing the uptake efficiency compared to that of the unmodified carriers (Figure 2C). To validate ATP-responsive drug release in the tumor cells, the intracellular ATP depletion assays significantly reduced nSDDP release compared to the ATP-replete controls, establishing a direct correlation between ATP concentration and payload release kinetics for metabolism-guided precision therapy (Figure 2D).

To establish cellular biocompatibility as a prerequisite for clinical translation, CCK-8 cytotoxicity assessment across multiple cell lines showed that nSDDP empty carriers induced minimal viability reduction after prolonged exposure, confirming exceptional cytocompatibility for therapeutic deployment (Figure 3A-3F). To assess systemic biosafety, the major organs underwent histological examination following extended nSDDP administration. The results showed that the myocardial fibers maintained parallel alignment, hepatic lobules displayed intact architecture, splenic red/white pulp boundaries remained distinct, pulmonary alveoli showed normal septation, and renal glomeruli exhibited preserved Bowman’s capsules. Meanwhile, both hematological indices and biochemical indices indicate that the nanocarrier is harmless to the organism and has good biosafety (Tables S1,S2). Collectively, these results confirmed the absence of pathological alterations at therapeutic doses (Figure 3G). The quantification of organ-selective distribution via in vivo imaging showed that the nSDDPs substantially reduced hepatic accumulation compared to the non-targeted MOFs, while significantly increasing pulmonary deposition (Figure 3H). Time-dependent fluorescence intensification peaked at 8 h post-injection, demonstrating a sustained lung-targeting capacity that concentrates chemotherapeutic payloads in tumors while markedly reducing off-target exposure. This dual-targeting strategy directly addresses DDP’s nephrotoxicity limitations via the spatial control of drug activation.

Figure 3 In vivo/vitro distribution and safety evaluation of nSDDPs. (A-F) Safety assessment of different concentrations of nSDDPs in different cell lines; (G) H&E staining for safety assessment of nSDDPs in different organs (10×, scale bar: 100 µm); (H) organ distribution of nSDDPs after injections at different times. Data are presented as the mean ± standard deviation (n=3). GC, Germ cells; H&E, hematoxylin and eosin; nSDDP, nanostructured sulforaphane and diamminedichloroplatinum (II) particle.

In vitro pharmacodynamic effect of the material

A549/DDP DDP-resistant cell lines represent clinically relevant models for investigating platinum resistance mechanisms in NSCLC. To verify the DDP resistance-relieving property of nSDDP, we first treated DDP-sensitive and DDP-resistant cell lines with different concentrations of DDP respectively. As shown in Figure 4A, A549/DDP cells exhibited certain resistance to DDP treatment. Meanwhile, when A549/DDP cells were treated with different concentrations of nSDDP, a significant contrast was observed in the inhibition rate compared to DDP-treated A549 cells, indicating that nSDDP possesses a remarkable ability to reverse DDP resistance—a conclusion further supported by the resistance index assay (Figure 4B). Notably, at 5-µM concentration, tumor cell inhibition was significantly increased compared to that of conventional DDP, while at 10-µM concentration, the efficacy was substantially improved. These results identified 5 µM as the optimal therapeutic concentration. The LDH release assays showed significantly increased membrane permeability in the nSDDP-treated cultures, while propidium iodide staining confirmed significantly elevated apoptotic cells compared to the DDP controls (Figure 4C). In the metastatic suppression analysis, scratch wound healing assays were employed to quantify anti-migratory efficacy. The nSDDP treatment significantly reduced wound closure compared to the DDP control treatment at 24 h, indicating the potent inhibition of cellular motility (Figure 4D).

Figure 4 In vitro experimental assessment of the anti-tumor efficacy of nSDDPs. (A) Cell inhibition rate assay of different doses of DDP and nSDDP on A549 and A549/DDP cell lines; (B) resistance index; (C) effect of the nSDDPs on A549/DDP LDH release; (D) effect of different doses of DDP on the migration ability of the A549/DDP cells (scale bar =10 µm); (E) effect of the nSDDPs on the proliferation rate of the A549/DDP cells (scale bar =10 µm); (F) effect of the nSDDPs on the apoptosis of the A549/DDP cells (scale bar =10 µm); (G) effects of different doses of DDP on the secretion levels of MMP-2, MMP-9, VEGF, and TGF-β in the A549/DDP cells; (H) effect of nSDDPs on the levels of AA, MDA, H₂O₂, TFR1, and tissue iron in the A549/DDP cells. Data are presented as the mean ± standard deviation (n=3). **, denotes a significant difference (P<0.01) between nSDDPs and DDP groups; ^##, indicates a significant difference (P<0.01) vs. the control group, while ^††, denotes a significant difference (P<0.01) between nSDDPs and DDP groups. Mito-Tracker dye was used for the indirect evaluation of cell proliferation, while the AV/PI staining method was employed to assess cell apoptosis. AA, arachidonic acid; AV/PI, annexin V/propidium iodide; Con, control; DAPI, 4',6-diamidino-2-phenylindole; DDP, diamminedichloroplatinum (II); LDH, lactate dehydrogenase; MDA, malondialdehyde; MMP, matrix metalloproteinase; nSDDP, nanostructured sulforaphane and diamminedichloroplatinum (II) particle; SFN, sulforaphane; TGF-β, transforming growth factor beta; VEGF, vascular endothelial growth factor.

The assessment of cell proliferation via Mito-Tracker immunofluorescence staining showed that the cellular staining intensity in the nSDDP-treated group was significantly lower compared to the cisplatin (DDP)-treated group (Figure 4E). This is consistent with the significant decrease in cell viability observed in the CCK-8 assay, confirming that nSDDP can sustainably inhibit the growth of tumor cells—likely associated with ferroptosis induction and mitochondrial damage triggered by nSDDP after cellular internalization. The apoptosis induction analysis revealed significantly enhanced cell death (Figure 4F). The complementary enzyme-linked immunosorbent assay (ELISA) quantification of metastatic regulators revealed coordinated pathway suppression. Specifically, MMP-9 secretion, VEGF, and TGF-β1 were all significantly decreased. This multi-factor inhibition disrupted the EMT cascade while impairing angiogenesis—key mechanisms underlying NSCLC dissemination (Figure 4G). Together, the results indicated caspase-9/3 cascade activation via mitochondrial cytochrome c release—a mechanism potentiated by targeted drug delivery to subcellular organelles.

The ferroptosis induction studies demonstrated the systematic disruption of redox homeostasis. The intracellular Fe2⁺ concentrations were significantly elevated with concomitant TFR1 upregulation, while AA accumulation significantly increased the lipid peroxidation markers MDA and H₂O₂. The parallel suppression of anti-oxidant defenses occurred through significantly inhibiting GPX4 activity. This triple-axis attack induced irreversible oxidative cascade amplification, specifically in the tumor cells (Figure 4H). The therapeutic synergy evaluation showed that the SFN co-administration significantly increased ferroptotic cell death compared to nSDDPs monotherapy. This occurred through the SFN-mediated inhibition of the xCT glutamate-cystine antiporter, which decreased the glutathione reserves required for GPX4 reactivation. The multi-modal therapeutic approach—which simultaneously targets apoptosis, metastatic pathways, proliferation signaling, and ferroptosis—collectively overcame the 7.3-fold DDP resistance index in A549/DDP cells. This represents a paradigm shift in platinum-refractory NSCLC management by addressing resistance through complementary biological mechanisms rather than dose escalation.

In vivo anti-tumor effect of the material

Animal models can be used to validate therapeutic efficacy against NSCLC chemoresistance. To quantify tumor suppression capabilities essential for clinical translation, subcutaneous xenograft studies measured volumetric changes in A549 tumor-bearing mice. The nSDDPs treatment led to a significantly greater reduction in the tumor growth coefficients compared to both DDP and SFN monotherapy, demonstrating superior control over platinum-resistant malignancies (Figure 5A). A histopathological assessment through H&E staining revealed that nSDDPs induced extensive tumor necrosis with significantly higher pyknotic nuclei density compared to the DDP groups, confirming potent apoptosis induction at the tissue level (Figure 5B). To evaluate metastatic regulation, an immunofluorescence analysis of invasion markers was conducted. The nSDDPs significantly decreased the MMP-2/9, TGF-β, and VEGF expression levels compared to the DDP group (Figure 5C). This coordinated biomarker modulation showed the comprehensive inhibition of invasion-angiogenesis cascades. The nSDDPs-treated tumors exhibited widespread mitochondrial shrinkage with characteristic cristae disintegration and membrane rupture (hallmarks of ferroptotic morphology), and most mitochondria (87.4%) retained structural integrity in the DDP group (Figure 5D). In the ferroptosis induction assessment, TEM was employed to visualize subcellular alterations (Figure 5E,5F). This visual evidence confirmed targeted oxidative organelle damage. The Western blot quantification of the ferroptosis regulators showed that the nSDDPs significantly suppressed GPX4 and SLC7A11 compared to DDP, establishing molecular pathway disruption (Figure 5G). The complementary qPCR analysis showed that the nSDDPs significantly decreased GPX4 mRNA, SLC7A11 mRNA, and FTH1 mRNA, while increasing ACSL4 mRNA (Figure 5H). This multi-omics verification demonstrated transcriptional reprogramming toward ferroptotic susceptibility. Therapeutic benchmarking confirmed the dual-mechanism advantage of the nSDDPs; DDP showed limited apoptosis induction, and SFN exhibited moderate ferroptosis; however, the nSDDPs achieved extensive apoptotic necrosis with near-universal mitochondrial destruction. This synergy overcomes chemoresistance by simultaneously activating caspase-dependent death and iron-mediated peroxidation cascades through tumor-targeted delivery.

Figure 5 In vivo experimental assessment of the anti-tumor efficacy of nSDDPs. (A) Detection results showing the effect of the nSDDPs on tumor growth in the A549 tumor-bearing mice; (B) H&E detection results showing the effect of the nSDDPs on tumors in the A549 tumor-bearing mice (20×, scale bar =50 µm); (C) immunofluorescence detection results showing the effect of the nSDDPs on the tumor MMPs, E-cadherin, TGF-β, and VEGF in the A549 tumor-bearing mice; (D) nSDDPs detection results for the mitochondrial ultrastructure in the A549 tumor-bearing mice, arrows in the figure indicate the locations of mitochondria; (E) schematic diagram of the relationship between cysteine transporters and ferroptosis; (F) effects of the nSDDPs on GPX4 enzyme activity in the A549/DDP cells; (G) nSDDPs detection results for the tumor ferroptosis-related proteins in the A549 tumor-bearing mice. (H) The mRNA expression of GPX4, SLC7A11, ACSL4 and FTH1. Data are presented as the mean ± standard deviation (n=3). ^##, indicates a significant difference (P<0.01) vs. the control group; while ^††, denotes a significant difference (P<0.01) between nSDDPs and DDP groups. Con, control; DDP, diamminedichloroplatinum (II); GSH, glutathione; H&E, hematoxylin and eosin; MMP, matrix metalloproteinase; nSDDP, nanostructured sulforaphane and diamminedichloroplatinum (II) particle; SFN, sulforaphane; TGF-β, transforming growth factor beta; VEGF, vascular endothelial growth factor.

Discussion

NSCLC is the most common type of lung cancer, accounting for approximately 85% of all lung cancer cases (4). Its high mortality rate is associated with late-stage diagnosis, rapid tumor progression, and the development of drug resistance. Although targeted therapy and immunotherapy have led to advancements in the treatment of NSCLC in recent years, chemotherapy remains the primary treatment modality for NSCLC, particularly in advanced patients. DDP, a classic platinum-based chemotherapy drug, is widely used in the clinical treatment of NSCLC. However, its clinical application faces numerous challenges, with side effects and drug resistance being the most prominent issues. Promoting ferroptosis can reverse DDP resistance and enhance the sensitivity of tumor cells to DDP. To overcome these difficulties in DDP treatment, this study developed a nanoparticle-based targeted delivery system. Compared with traditional DDP treatments, nanoparticles can deliver drugs precisely to tumor cells through targeted action, significantly increasing drug concentration in the tumor area and thereby enhancing its anti-cancer effect. Compared with previously reported MOF-based or lipid-based DDP nanocarriers, our nSDDPs exhibit notable performance advantages in several key aspects: EGFR-specific targeting modification enables more precise tumor targeting and accumulation than most non-targeted or single-responsive carriers, alleviating the key limitation of insufficient targeting in traditional carriers; its excellent surface modifiability outperforms some MOF materials with rigid structures and high functionalization difficulty, facilitating subsequent functional expansion; the relatively simplified synthesis process reduces preparation costs and enhances translational potential; and the unique ATP/pH dual-responsive release design is more compatible with the dual physicochemical characteristics of the tumor microenvironment compared to carriers with only single pH responsiveness, allowing precise control of drug release at the tumor site, minimizing damage to normal tissues, and further enhancing the specificity and safety of treatment.

Small-sized nanoparticles have significant advantages in NSCLC treatment, as they address the limitations of traditional drug treatments and provide targeted treatment and multifunctionality. Due to their extremely small size, nanoparticles can better penetrate tumor cells and concentrate in the TME, increasing local drug concentration and reducing toxicity to normal tissues. Additionally, nanoparticles can be surface-modified to achieve targeted functionality, enhancing drug selective delivery and improving treatment outcomes. In the treatment of NSCLC, nanoparticles can effectively enhance the efficacy of chemotherapy drugs, overcome drug resistance, reduce side effects, and exert synergistic effects by combining different drugs through synthetic complexes or carrier systems. This study successfully synthesized nSDDPs with a particle size of approximately 110 nm, exhibiting long-term storage stability and particle size uniformity. Notably, the synthesized nSDDPs maintained structural integrity after EGFR modification and DL, with stable crystalline properties that ensure reliable drug release performance, while their excellent colloidal stability prevents aggregation during circulation and storage. nSDDPs have distinct pH- and ATP-responsive release characteristics, facilitating controlled drug release in the TME. This dual-responsive design enables specific drug release only in the acidic and high-ATP tumor microenvironment, avoiding premature drug leakage in normal tissues and thus minimizing systemic toxicity, which solves the key problem of non-specific distribution of traditional DDP.

Nanoparticles designed using the antibody-receptor concept have significant specificity advantages, notably including an ability to precisely identify and target lung tumor cells. By modifying antibodies or their fragments (e.g., monoclonal antibodies) onto the surface of nanomaterials, they can bind to receptors specific to lung cancer cells (e.g., EGFR and VEGF receptors), thereby achieving selective recognition and binding to tumor cells. This targeting significantly increases drug concentration in tumor tissues, enhancing therapeutic efficacy while minimizing damage to healthy tissues. Additionally, antibody-receptor-targeted nanomaterials can overcome biological barriers, enhance drug penetration, and stably release drugs in the TME, significantly improving treatment precision and efficiency. Thus, this design not only enhances the targeting and specificity of lung cancer treatments but may also reduce the side effects associated with conventional therapies. This study successfully synthesized nanoparticles to target NSCLC tumors. The EGFR-targeted modification of nSDDPs significantly improves their uptake efficiency by NSCLC cells, and the endocytosis pathway mediated by lipid rafts and dynamin further enhances the specific accumulation of nanoparticles in tumor cells. Additionally, these nanoparticles did not accumulate significantly in normal tissues, greatly reducing the toxic side effects of traditional DDP formulations. In vivo distribution studies further confirmed that nSDDPs preferentially accumulate in lung tumors while reducing deposition in non-target organs such as the liver, which significantly mitigates the off-target toxicity issue of traditional chemotherapy drugs.

Nanoparticles offer significant advantages in terms of their safety and long-term stable dispersion in medical applications. First, due to their small size and large surface area, nanoparticles can effectively reduce non-specific interactions with healthy tissues, thereby lowering toxicity. Additionally, through precise surface modification (e.g., surface hydrophilization or PEGylation), their biocompatibility can be enhanced, reducing immune system rejection and thereby improving safety. Second, the long-term stable dispersion of nanoparticles is crucial for ensuring their effective circulation and targeting in the body. By optimizing the surface properties of nanoparticles, their aggregation and precipitation in blood or tissues can be effectively avoided, ensuring the stable dispersion of nanoparticles in the body, prolonging the half-life of drugs, thereby enhancing therapeutic efficacy and reducing side effects. Preliminary safety evaluations indicated that nSDDPs have favorable biocompatibility: empty carriers showed no significant cytotoxicity to multiple normal and tumor cell lines, and long-term in vivo administration did not cause pathological damage to major organs such as the heart, liver, spleen, kidneys, and lungs. Histological section results from various tissues and organs indicated that the nSDDPs did not exert significant toxic effects on these tissues and organs, providing a solid foundation for further pharmacodynamic evaluation. This study employed a subcutaneous tumor-bearing model primarily based on two key advantages: first, its simple operation and high reproducibility enable standardized comparison of pharmacodynamic data between groups, reducing experimental errors caused by individual differences; second, as a classic platform for initial pharmacological research, it allows rapid verification of the nanocarrier’s targeting ability, drug release, and core anti-tumor functions, laying a solid foundation for subsequent in-depth studies using orthotopic or metastatic tumor models. It should be noted that the subcutaneous tumor model does not fully mimic the tumor microenvironment and metastatic characteristics of clinical NSCLC, and the related therapeutic efficacy needs further verification in more clinically relevant models.

Although the nSDDPs in this study exhibited favorable biocompatibility and targeted anti-tumor efficacy, the clinical translation of MOF-based therapies still faces well-recognized inherent challenges in the field. First, Good Manufacturing Practice (GMP)-compliant large-scale production requires further optimization of process parameters and establishment of a strict quality control system to ensure the stability and consistency of products across different batches; the simplified synthetic route of this study has laid a foundation for subsequent process scaling-up. Second, the long-term immunogenicity of MOF materials needs to be clarified by extending preclinical observation periods and conducting more comprehensive immunocompatibility assessments to avoid non-specific immune responses induced by long-term in vivo circulation. Additionally, the in vivo metabolic pathways of MOF carriers and the safety of their degradation products require further elucidation. Addressing these issues through subsequent targeted preclinical studies will be crucial to ensure the feasibility of translating the nSDDPs system from the laboratory to the clinic.

The A549/DDP drug-resistant cell line is widely used to study the mechanisms of tumor drug resistance, particularly the mechanisms of cellular resistance to DDP, such as oxidative stress, drug efflux, and DNA repair pathways. Additionally, these cell lines are used in drug development and screening to evaluate the therapeutic efficacy of novel drugs or drug combinations against DDP-resistant tumors. We found that the nSDDPs exhibited a more significant inhibitory effect in the A549/DDP cell line than DDP. This superior inhibitory effect is attributed to nSDDPs’ ability to bypass drug efflux-mediated resistance mechanisms through targeted delivery, thereby increasing intracellular drug accumulation. Furthermore, promoting ferroptosis by enhancing intracellular iron accumulation and reactive oxygen species levels effectively reversed DDP resistance and enhanced the cytotoxic effect on tumor cells. Specifically, nSDDPs induce ferroptosis by regulating key molecules in iron metabolism, lipid peroxidation, and antioxidant pathways, which complements the DNA-damaging effect of DDP and forms a synergistic anti-tumor effect. Combining ferroptosis inducers with DDP in combination therapy has also been shown to effectively overcome tumor cell resistance, enhance the efficacy of DDP, and provide new strategies for cancer treatment. This study innovatively combines SFN, a natural ferroptosis inducer, with DDP through a nano-delivery system, achieving synchronized delivery and synergistic action to overcome DDP resistance, representing a practical improvement compared to single-agent therapy.

During ferroptosis, AA exacerbates ferroptosis by promoting lipid peroxidation. GPX4 exerts a protective effect by preventing the accumulation of lipid peroxidation products. The inhibition of GPX4 enzyme activity directly induces ferroptosis. Cysteine transporters exert an anti-oxidant effect by maintaining intracellular glutathione synthesis, thereby inhibiting the exacerbation of oxidative stress and reducing ferroptosis. We found that the combination of SFN and DDP effectively enhanced the cytotoxic effect of DDP on tumor cells by inducing ferroptosis, thereby increasing cellular sensitivity to DDP. In terms of tumor size, histological changes, and metastatic factors, the nSDDPs group demonstrated significantly superior tumor suppression effects compared to the DDP group, with more effective control of tumor growth. This includes not only inhibiting tumor proliferation but also suppressing tumor migration and invasion by regulating the expression of metastasis-related factors, achieving comprehensive tumor growth control. Additionally, pathological changes in the tumor tissue showed reduced invasiveness and metastatic potential. Furthermore, in the detection of iron death-related indicators, the nSDDPs group demonstrated higher activation effects in promoting the expression of key proteins and genes associated with iron death (e.g., GPX4 and FTH1), significantly enhancing tumor cell mortality, and showing better efficacy in overcoming DDP resistance and in anti-tumor therapy. The multi-omics analysis further confirms that nSDDPs regulate the transcriptional and translational levels of ferroptosis-related molecules, providing a clear molecular mechanism for their ability to overcome DDP resistance. This suggests that by effectively promoting ferroptosis, nSDDPs enhance the therapeutic efficacy of DDP, offering a more promising treatment option for NSCLC.

Conclusions

This study systematically explored nSDDPs, successfully synthesizing and characterizing the nanoparticles, and confirming their excellent targeting ability and controllable responsive drug release performance. The cell-level studies showed that the nSDDPs not only exhibited high safety but also significantly suppressed the migration and proliferation of A549/DDP cells, induced apoptosis, and effectively overcame DDP resistance. Animal experiments further verified that the nSDDPs significantly inhibited tumor growth in tumor-bearing mice without causing obvious toxicity to major organs. Our research findings provide new insights into the treatment of NSCLC. nSDDPs are expected to become a new type of nano-preparation with high efficiency, low toxicity, and the ability to overcome drug resistance. Our findings offer important scientific evidence and practical directions for drug development and combination therapy strategies in the field of cancer treatment.

Acknowledgments

None.

Footnote

Reporting Checklist: The authors have completed the ARRIVE and MDAR reporting checklists. Available at https://tlcr.amegroups.com/article/view/10.21037/tlcr-2025-1123/rc

Data Sharing Statement: Available at https://tlcr.amegroups.com/article/view/10.21037/tlcr-2025-1123/dss

Peer Review File: Available at https://tlcr.amegroups.com/article/view/10.21037/tlcr-2025-1123/prf

Funding: This work was supported by the Haiyan Research Fund Youth Project of Harbin Medical University Cancer Hospital (Nos. JJQN2023-03 and JJQN2023-06).

Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at https://tlcr.amegroups.com/article/view/10.21037/tlcr-2025-1123/coif). The authors have no conflicts of interest to declare.

Ethical Statement: The authors are accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved. All the animal experiments were conducted in compliance with institutional guidelines for the care and use of animals, and were approved by the Institutional Research Ethics Committee of Harbin Medical University Cancer Hospital (No. KY2022-71, 2022/11/3).

Open Access Statement: This is an Open Access article distributed in accordance with the Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License (CC BY-NC-ND 4.0), which permits the non-commercial replication and distribution of the article with the strict proviso that no changes or edits are made and the original work is properly cited (including links to both the formal publication through the relevant DOI and the license). See: https://creativecommons.org/licenses/by-nc-nd/4.0/.

References

1. Jeon H, Wang S, Song J, et al. Update 2025: Management of Non Small-Cell Lung Cancer. Lung 2025;203:53. [Crossref] [PubMed]
2. Mithoowani H, Febbraro M. Non-Small-Cell Lung Cancer in 2022: A Review for General Practitioners in Oncology. Curr Oncol 2022;29:1828-39. [Crossref] [PubMed]
3. Huang Q, Li Y, Huang Y, et al. Advances in molecular pathology and therapy of non-small cell lung cancer. Signal Transduct Target Ther 2025;10:186. [Crossref] [PubMed]
4. De Lucia A, Mazzotti L, Gaimari A, et al. Non-small cell lung cancer and the tumor microenvironment: making headway from targeted therapies to advanced immunotherapy. Front Immunol 2025;16:1515748. [Crossref] [PubMed]
5. Remon J, Soria JC, Peters S, et al. Early and locally advanced non-small-cell lung cancer: an update of the ESMO Clinical Practice Guidelines focusing on diagnosis, staging, systemic and local therapy. Ann Oncol 2021;32:1637-42. [Crossref] [PubMed]
6. Pennell NA, Arcila ME, Gandara DR, et al. Biomarker Testing for Patients With Advanced Non-Small Cell Lung Cancer: Real-World Issues and Tough Choices. Am Soc Clin Oncol Educ Book 2019;39:531-42. [Crossref] [PubMed]
7. Guo LL, Yang XH, Jin HZ, et al. TFAP2A induces cisplatin resistance via BNIP3-mediated mitophagy in non-small cell lung cancer. Transl Cancer Res 2025;14:5353-69. [Crossref] [PubMed]
8. Raziq MF, Manzoor H, Kayani MUR. Non-small Cell Lung Cancer, Immunotherapy and the Influence of Gut Microbiome. Curr Microbiol 2025;82:419. [Crossref] [PubMed]
9. Nafees B, Lloyd AJ, Dewilde S, et al. Health state utilities in non-small cell lung cancer: An international study. Asia Pac J Clin Oncol 2017;13:e195-203. [Crossref] [PubMed]
10. Nagano T, Tachihara M, Nishimura Y. Molecular Mechanisms and Targeted Therapies Including Immunotherapy for Non-Small Cell Lung Cancer. Curr Cancer Drug Targets 2019;19:595-630. [Crossref] [PubMed]
11. Patel SA, Weiss J. Advances in the Treatment of Non-Small Cell Lung Cancer: Immunotherapy. Clin Chest Med 2020;41:237-47. [Crossref] [PubMed]
12. Hua P, Olofson A, Farhadi F, et al. Predicting targeted therapy resistance in non-small cell lung cancer using multimodal machine learning. J Thorac Dis 2025;17:8700-14. [Crossref] [PubMed]
13. Zhang Y, Zhang W, Zhao Y, et al. Bioactive sulforaphane from cruciferous vegetables: advances in biosynthesis, metabolism, bioavailability, delivery, health benefits, and applications. Crit Rev Food Sci Nutr 2025;65:3027-47. [Crossref] [PubMed]
14. Alhazmi N, Subahi A. Impact of Sulforaphane on Breast Cancer Progression and Radiation Therapy Outcomes: A Systematic Review. Cureus 2025;17:e78060. [Crossref] [PubMed]
15. Plafker KS, Georgescu C, Pezant N, et al. Sulforaphane acutely activates multiple starvation response pathways. Front Nutr 2024;11:1485466. [Crossref] [PubMed]
16. Kuran D, Pogorzelska A, Wiktorska K. Breast Cancer Prevention-Is there a Future for Sulforaphane and Its Analogs? Nutrients 2020;12:1559. [Crossref] [PubMed]
17. Xagoraris I, Yang Y, Bougka E, et al. Sulforaphane promotes natural killer cell-mediated anti-tumor immune responses partially via cGAS-STING pathway in classical Hodgkin lymphoma. Leukemia 2025;39:1787-90. [Crossref] [PubMed]
18. Traka MH, Melchini A, Mithen RF. Sulforaphane and prostate cancer interception. Drug Discov Today 2014;19:1488-92. [Crossref] [PubMed]
19. Russo M, Spagnuolo C, Russo GL, et al. Nrf2 targeting by sulforaphane: A potential therapy for cancer treatment. Crit Rev Food Sci Nutr 2018;58:1391-405. [Crossref] [PubMed]
20. Liu P, Zhang B, Li Y, et al. Potential mechanisms of cancer prevention and treatment by sulforaphane, a natural small molecule compound of plant-derived. Mol Med 2024;30:94. [Crossref] [PubMed]
21. Li W, Trieu J, Blazev R, et al. Sulforaphane attenuates cancer cell-induced atrophy of C2C12 myotubes. Am J Physiol Cell Physiol 2023;324:C205-21. [Crossref] [PubMed]
22. Masoom M, Khan MA. Efficacy of sulforaphane in skin cancer animal models: A systematic review. Polim Med 2024;54:105-11. [Crossref] [PubMed]
23. Rafiei H, Ashrafizadeh M, Ahmadi Z. MicroRNAs as novel targets of sulforaphane in cancer therapy: The beginning of a new tale? Phytother Res 2020;34:721-8. [Crossref] [PubMed]
24. Gu HF, Mao XY, Du M. Metabolism, absorption, and anti-cancer effects of sulforaphane: an update. Crit Rev Food Sci Nutr 2022;62:3437-52. [Crossref] [PubMed]
25. Dasari S, Tchounwou PB. Cisplatin in cancer therapy: molecular mechanisms of action. Eur J Pharmacol 2014;740:364-78. [Crossref] [PubMed]
26. Dasari S, Njiki S, Mbemi A, et al. Pharmacological Effects of Cisplatin Combination with Natural Products in Cancer Chemotherapy. Int J Mol Sci 2022;23:1532. [Crossref] [PubMed]
27. Zoń A, Bednarek I. Cisplatin in Ovarian Cancer Treatment-Known Limitations in Therapy Force New Solutions. Int J Mol Sci 2023;24:7585. [Crossref] [PubMed]
28. Freyer DR, Brock PR, Chang KW, et al. Prevention of cisplatin-induced ototoxicity in children and adolescents with cancer: a clinical practice guideline. Lancet Child Adolesc Health 2020;4:141-50. [Crossref] [PubMed]
29. Ghosh S. Cisplatin: The first metal based anticancer drug. Bioorg Chem 2019;88:102925. [Crossref] [PubMed]
30. Makovec T. Cisplatin and beyond: molecular mechanisms of action and drug resistance development in cancer chemotherapy. Radiol Oncol 2019;53:148-58. [Crossref] [PubMed]
31. Mu Y, Dong Y, Zheng M, et al. Identification of a prognostic gene signature in patients with cisplatin resistant squamous cell lung cancer. J Thorac Dis 2024;16:4567-83. [Crossref] [PubMed]
32. Gandin V, Hoeschele JD, Margiotta N. Special Issue "Cisplatin in Cancer Therapy: Molecular Mechanisms of Action 3.0". Int J Mol Sci 2023;24:7917. [Crossref] [PubMed]
33. Wang S, Liu G, Yu L, et al. Fluorofenidone enhances cisplatin efficacy in non-small cell lung cancer: a novel approach to inhibiting cancer progression. Transl Lung Cancer Res 2024;13:3175-88. [Crossref] [PubMed]
34. Berkel C, Cacan E. Estrogen- and estrogen receptor (ER)-mediated cisplatin chemoresistance in cancer. Life Sci 2021;286:120029. [Crossref] [PubMed]
35. Sun CY, Zhang QY, Zheng GJ, et al. Phytochemicals: Current strategy to sensitize cancer cells to cisplatin. Biomed Pharmacother 2019;110:518-27. [Crossref] [PubMed]
36. Desilets A, Adam JP, Soulières D. Management of cisplatin-associated toxicities in bladder cancer patients. Curr Opin Support Palliat Care 2020;14:286-92. [Crossref] [PubMed]
37. Wu MX, Yang YW. Metal-Organic Framework (MOF)-Based Drug/Cargo Delivery and Cancer Therapy. Adv Mater 2017; [Crossref]
38. Sun L, Liu H, Ye Y, et al. Smart nanoparticles for cancer therapy. Signal Transduct Target Ther 2023;8:418. [Crossref] [PubMed]
39. Zhao D, Zhang W, Yu S, et al. Application of MOF-based nanotherapeutics in light-mediated cancer diagnosis and therapy. J Nanobiotechnology 2022;20:421. [Crossref] [PubMed]
40. Yang J, Dai D, Zhang X, et al. Multifunctional metal-organic framework (MOF)-based nanoplatforms for cancer therapy: from single to combination therapy. Theranostics 2023;13:295-323. [Crossref] [PubMed]
41. Wang Q, Yu Y, Chang Y, et al. Fluoropolymer-MOF Hybrids with Switchable Hydrophilicity for (19)F MRI-Monitored Cancer Therapy. ACS Nano 2023;17:8483-98. [Crossref] [PubMed]
42. Dou Y, Wang Y, Tian S, et al. Metal-organic framework (MOF)-based materials for pyroptosis-mediated cancer therapy. Chem Commun (Camb) 2024;60:6476-87. [Crossref] [PubMed]
43. Cai J, Xu Y, Liao F. Advances in multifunctional metal-organic framework (MOF)-based nanoplatforms for cancer starvation therapy. Expert Rev Mol Med 2024;26:e27. [Crossref] [PubMed]
44. Mohan B, Modi K, Singh G, et al. Understanding the Electrochemical MOF Sensors in Detecting Cancer with Special Emphasis on Breast Carcinoma Biomarkers. Top Curr Chem (Cham) 2025;383:9. [Crossref] [PubMed]
45. Du H, Meng S, Geng M, et al. Detachable MOF-Based Core/Shell Nanoreactor for Cancer Dual-Starvation Therapy With Reversing Glucose and Glutamine Metabolisms. Small 2023;19:e2303253. [Crossref] [PubMed]
46. Zhang C, Xin L, Li J, et al. Metal-Organic Framework (MOF)-Based Ultrasound-Responsive Dual-Sonosensitizer Nanoplatform for Hypoxic Cancer Therapy. Adv Healthc Mater 2022;11:e2101946. [Crossref] [PubMed]

(English Language Editor: L. Huleatt)