Induction Of Apoptosis In Cancer Cells Research Articles

Polymer-based nanodrugs enhance sonodynamic therapy through epigenetic reprogramming of the immunosuppressive tumor microenvironment.

While sonodynamic therapy (SDT) has shown promise in treating triple-negative breast cancer (TNBC) due to its non-invasive nature, deep tissue penetration, and induction of immunogenic cell death (ICD), its efficacy remains limited by the complex immunosuppressive tumor microenvironment (TME). In this study, we developed tumor microenvironment-responsive nanoparticles (GdNPs) to enhance SDT effectiveness through epigenetic reprogramming of the TME by encapsulating the sonosensitizer chlorin e6 (Ce6) and the histone deacetylase 6 (HDAC6) inhibitor Ricolinostat (Ric) (GdNPs/Ce6-Ric). GdNPs/Ce6-Ric effectively accumulate at tumor sites via the enhanced permeability and retention (EPR) effect and release Ce6 and Ric in response to the acidic TME. Upon ultrasound stimulation, GdNPs/Ce6-Ric induce cancer cell apoptosis and trigger ICD by generating reactive oxygen species (ROS), which activate cytotoxic T cells and promote tumor cell elimination. Notably, the epigenetic modulation by Ric within the immunosuppressive TME increased the proportion of natural killer (NK) cells and cytotoxic T cells while decreasing the population of immunosuppressive regulatory T (Treg) cells. This modulation synergistically enhanced the anti-tumor effects of SDT by downregulating the HDAC6/p-STAT3/PD-L1 pathway. Furthermore, GdNPs/Ce6-Ric minimized lung metastases by not only improving systemic immune responses but also inhibiting TGFβ-induced epithelial-mesenchymal transition (EMT) of tumor cells through the blockade of α-tubulin deacetylation. Thus, GdNPs/Ce6-Ric-based epigenetic modulation of the immunosuppressive TME offers a promising approach to enhance the efficacy of SDT in treating TNBC.

Application of MMP-2-responsive in situ forming injectable hydrogel in preventing the recurrence of oral squamous cell carcinoma.

Oral squamous cell carcinoma is one of the most common types of cancer. Surgical resection is one of the most important treatments at present. However, patients often suffer from regional recurrence after surgery. Therefore, new strategies for combination therapy need to be investigated. This work identified a smart injectable hydrogel system co-delivering DOX and sunitinib nanoparticles. The nano drugs are continuously released from the hydrogels and effectively taken up by cells. The nano drugs in the hydrogel can inhibit tumor cell viability and induce cancer cell apoptosis in vitro. The drug-loaded hydrogel could control the recurrence of subcutaneous xenograft tumors, prolong the survival time, and have no obvious toxicity in nude mice. These findings suggest that this smart injectable hydrogel system may provide new ideas for the comprehensive treatment of oral squamous cell carcinoma.

Proximal Oblique‐Packing of Heptamethine Cyanines through Spiro‐Connection Boosts Triplet State Generation in Near‐Infrared

Near‐infrared (NIR) triplet dyes are the cornerstones of cutting‐edge biomedical and material applications. The difficulty in rational development of triplet dyes increases exponentially as the absorption wavelength shifts deeper into the NIR range. Although classical H‐/J‐typed packing of NIR dyes has the potential to enhance intersystem crossing (ISC) compared with that in single‐chromophore dyes, the triplet state quantum yields remain limited in such strategy. Herein, proximal oblique‐packed (V‐shaped) heptamethine cyanines (SZ780) through spiro‐connection were achieved. Multi‐channel ultrafast ISC were direct observed in SZ780 and a record high ISC rate constant (up to ~1011 s‐1) is registered among all the reported NIR triplet dyes. SZ780 exhibits a triplet state quantum yield of 18.9% upon excitation at 750 nm, which is almost an order of magnitude higher than that of the monomer (IR780, 2.1%) and nearly threefold increase compared to that of the H‐packed dimer (SC780) (6.7%). Moreover, SZ780 efficiently generates singlet oxygen under 808 nm light irradiation, inducing cancer cell apoptosis in vivo. These findings demonstrate that constructing V‐aggregated dyes system by spiro‐connection offers a powerful approach for the design of high‐performance NIR triplet sensitizers.

Proximal Oblique-Packing of Heptamethine Cyanines through Spiro-Connection Boosts Triplet State Generation in Near-Infrared.

Near-infrared (NIR) triplet dyes are the cornerstones of cutting-edge biomedical and material applications. The difficulty in rational development of triplet dyes increases exponentially as the absorption wavelength shifts deeper into the NIR range. Although classical H-/J-typed packing of NIR dyes has the potential to enhance intersystem crossing (ISC) compared with that in single-chromophore dyes, the triplet state quantum yields remain limited in such strategy. Herein, proximal oblique-packed (V-shaped) heptamethine cyanines (SZ780) through spiro-connection were achieved. Multi-channel ultrafast ISC were direct observed in SZ780 and a record high ISC rate constant (up to ~1011 s-1) is registered among all the reported NIR triplet dyes. SZ780 exhibits a triplet state quantum yield of 18.9% upon excitation at 750 nm, which is almost an order of magnitude higher than that of the monomer (IR780, 2.1%) and nearly threefold increase compared to that of the H-packed dimer (SC780) (6.7%). Moreover, SZ780 efficiently generates singlet oxygen under 808 nm light irradiation, inducing cancer cell apoptosis in vivo. These findings demonstrate that constructing V-aggregated dyes system by spiro-connection offers a powerful approach for the design of high-performance NIR triplet sensitizers.

Roles and therapeutic opportunities of ω-3 long-chain polyunsaturated fatty acids in lung cancer.

Over the past decades, researchers have continuously investigated the potential functions of long-chain polyunsaturated fatty acids (LCPUFAs) in cancers, including lung cancer. The ω-3 LCPUFAs, primarily consisting of eicosapentaenoic acid and docosahexaenoic acid, were found to modify inflammatory tumor microenvironment, induce cancer cell apoptosis and autophagy, and suppress tumor development when administered alone or with other therapeutical strategies. Although the precise anti-tumor mechanism has not been elucidated yet, ω-3 LCPUFAs are often used in the nutritional treatment of patients with cancer due to their ability to significantly improve patient's nutritional status, increase the sensitivity of tumor cells to treatments, and alleviate cancer-related complications. Here we present the key roles of ω-3 LCPUFAs as dietary supplementations in lung cancer, comprehensively review the recent progress on the underlying mechanisms of cancer cell regulation by ω-3 LCPUFAs, and introduce the application of ω-3 LCPUFAs in the clinical management of lung cancer and its malignant complications.

The development of α, β-unsaturated lactam-based andrographolide derivatives as anti-gastric cancer agents with the ability of inhibiting the ERK/c-Fos/Jun pathway.

Gastric cancer remains one of the global health threats for human beings. However, the therapeutic efficacy of the widely-used chemotherapy is usually limited due to the lack of specificity and the related toxicity. Only limited therapeutic agents were demonstrated to show selective and potent inhibitory activity to gastric cancer cells. In this study, we report the first α, β-unsaturated lactam-based andrographolide derivative P16 with the ability to potently and selectively inhibit the proliferation and migration of gastric cancer cells MGC-803. Moreover, the in vivo studies showed that P16 exhibited remarking anti-gastric cancer activity by significantly reducing the growth of tumor without losing the body weight. Further anticancer mechanistic studies indicated that P16 exerted its potent and selective anti-gastric cancer effect by arresting cell cycle at G2/M phase and inducing cancer cell apoptosis through intrinsic mitochondria-mediated pathway. Notably, for the first time, we found that andrographolide derivative P16 could reduce the activities of the ERK/c-Fos/Jun pathway to exert the anti-gastric cancer efficiency. This is the first time to reveal the novel role of ERK/c-Fos/Jun signaling in andrographolide derivative-mediated anti-gastric cancer processes. Overall, derivative P16 represents a valuable candidate for new therapeutic agent discovery in gastric cancer chemotherapy. In addition, pharmacological characterizations of derivative P16, together with another 33 new semi-synthesized andrographolide derivatives, provides a systematic structure-activity relationship (SAR) analysis for this class of compounds, elucidating useful information on structural requirements for potent and selective anti-gastric cancer inhibition.

Global research on nanomaterials for liver cancer from 2004 to 2023: a bibliometric and visual analysis.

Primary liver cancer, particularly hepatocellular carcinoma, is one of the most common gastrointestinal cancers. An increasing number of studies indicate that nanomaterials play a significant role in the diagnosis and treatment of liver cancer. However, despite the extensive and diverse research on nanomaterials and liver cancer, bibliometric studies in this field have not yet been reported. This study aims to comprehensively evaluate the application prospects and development trends of nanomaterials in primary liver cancer over the past 20years. By elucidating the current state of research on liver cancer, we intend to provide valuable reference information for researchers in this field. We conducted a comprehensive search of the Web of Science Core Collection for publications related to liver cancer and nanomaterials from January 1, 2004, to December 31, 2023. Relevant literature was selected based on specific inclusion and exclusion criteria. These selected publications were subsequently analyzed using CiteSpace, VOSviewer, and the R package "bibliometrix" to identify trends, influential countries, institutions, authors, journals, and research hotspots in this field. This study included a total of 1641 publications, with an annual growth rate of 25.45%. China and the United States are leading in this field, accounting for 67.46% and 11.27% of the total publications, respectively. The Chinese Academy of Sciences and Shao D are the most cited institution and author, respectively. The International Journal of Nanomedicine is the most influential journal in this field, while Biomaterials is the most highly cited and co-cited journal. Research hotspots mainly focus on improving drug delivery efficiency, inducing cancer cell apoptosis, photodynamic therapy, photothermal therapy, and combination treatments. Emerging research directions include the tumor microenvironment, polyethylene glycol, and immunogenic cell death. The results of this study indicate that the application of nanomaterials in the field of liver cancer is gradually becoming a significant research area, with a focus on improving drug delivery efficiency, enhancing therapeutic efficacy, and reducing side effects.

Vanadium Complexes for Mitochondria-Targeted Photodynamic Therapy.

This minireview focuses on vanadium complexes in photodynamic therapy (PDT), particularly for their potential as mitochondria-targeted anticancer agents. Vanadium's coordination versatility supports its bioactivity, showing promise in insulin-mimetic, lipid-lowering, and antitumor effects. PDT leverages these complexes' redox properties, producing reactive oxygen species (ROS) within mitochondria to induce cancer cell apoptosis with minimal impact on healthy cells. The review covers design strategies to improve mitochondrial localization, photodynamic efficiency, and selective cytotoxicity while addressing challenges like photostability and targeting for next-gen PDT applications.

Mechanisms and Applications of AZD6738 on Breast Cancer

Breast cancer (BC) is a cancer that is prevalent in women. However, the heterogeneity of BC in patients, especially triple-negative breast cancer (TNBC), leads to cases with poor outcomes, which requires more efficient treatment approaches. AZD6738, also known as ceralasertib, a selective inhibitor of Ataxia telangiectasia and Rad3-related (ATR) kinase, can be used for cancer treatment. In clinical practice, it is often used in combination with other drugs or chemotherapy to enhance treatment efficacy. ATR kinase is activated during DNA damage, affecting the cell cycle and responding to DNA replication stress (RS) by activating CHK1 and a series of downstream reactions. AZD6738 can block the ATR pathway by inhibiting the activation of CHK1, thereby inducing cancer cell apoptosis. Moreover, researchers conducted clinical studies on AZD6738 and explored the efficacy of its combination therapy with other drugs, such as PARP inhibitor olaparib. This review focuses on the structure and function of AZD6738, as well as its applications in TNBC clinical trials. Future studies of AZD6738 are required to further explore the mechanisms and clinical applications in the treatment of breast cancer and other cancers.

Research Progress of Endoplasmic Reticulum Targeting Metal Complexes in Cancer Therapy.

The development of anticancer drugs that target different organelles has received extensive attention due to the characteristics of cancer recurrence, metastasis, and drug resistance. The endoplasmic reticulum (ER) is an important structure within the cell that is primarily responsible for protein synthesis, folding, modification, and transport and plays a crucial role in cell function and health. ER stress activation induces cancer cell apoptosis. New anticancer drugs with different anticancer mechanisms and selectivity can be designed because of redox activity, composition diversity, and metal complexes structure regulation. Over the past few decades, dozens of metal complexes have killed cancer cells through ER stress, showing powerful tumor-suppressive effects. This review summarizes the progress of research on anticancer metallic drugs that induce ER stress over the past few years, which is expected to bring more breakthroughs in the field of medicine and life science.

PEG-BCT-100 and Canavanine Synergistically Induce Apoptosis in Arginine Biosynthetic Enzyme-Deficient Pancreatic Cancer.

This study investigates the synergistic antitumor effect of PEG-BCT-100, an arginase, in clinical trials, with canavanine in pancreatic cancer, in vitro and in vivo. The treatment induces cancer cell apoptosis while sparing normal fibroblasts. Our findings suggest heightened susceptibility of pancreatic tumors deficient in arginine biosynthesis enzymes ASS1 and OTC.

Isolation, purification, structural characterization, and antitumor activity of Gynura divaricata polysaccharides.

In this study, a water-soluble, homogeneous polysaccharide, GDPs-1, was isolated from Gynura divaricata and purified. It was subsequently subjected to structural characterization and biological activity assessment. Structural characterization revealed that GDPs-1 was mainly composed of glucose, galactose, and arabinose, with a molecular weight of 55.1 kDa. Its main structural backbone was →[4)-α-D-Glcp-(1]22 → 4)-α-D-Glcp-(1 → 4)-α-D-Glcp-(1 → [4)-β-D-Galp-(1]11→, with two additional branched structures. In vitro experiments demonstrated that GDPs-1 induced cancer cell apoptosis, caused cell cycle blockade, increased the Bax/Bcl-2 protein ratio, attenuated epithelial-mesenchymal transition, and activated caspase-3 protein in carcinoma cells, thereby exerting antitumor effects on tumor cells without affecting the growth of normal cells. Therefore, GDPs-1 may be further explored as a novel medicine to treat lung and liver cancer.

Erianin inhibits the progression of DDP-resistant lung adenocarcinoma by regulating the Wnt/β-catenin pathway and activating the caspase-3 for apoptosis in vitro and in vivo

BackgroundPlatinum-based chemotherapy is one of the main treatments for lung adenocarcinoma (LUAD). However, the toxic side effects and drug resistance of chemotherapeutic drugs on normal cells are still a thorny problem in clinical treatment. Dendrobium is one of the three largest genera of Orchidaceous family, which has ornamental and medicinal value. Dendrobium is mainly distributed in the tropics and subtropics of South Asia, Oceania and other regions, with 1547 species of Dendrobium currently known. In China, “Shi hu” and “Tie pi shi hu” are well-known traditional medicines and have been included in the Chinese Pharmacopoeia (Editorial Board of Chinese Pharmacopoeia, 2020). Erianin is a natural product isolated from Dendrobium and is considered as a potential anticancer molecule due to its remarkable anti-tumor effects through various mechanisms, among which induced cancer cell apoptosis, inhibited invasion and migration. This study preliminarily explored the mechanism of Erianin inhibiting the progression of cisplatin (DDP) resistant LUAD in vivo and in vitro.MethodsThe effect of Erianin on the proliferation of DDP-resistant LUAD cells was detected by CCK-8 assay, wound healing assay and cloning assay. Transwell assay was used to evaluate the effect of Erianin on cell invasion and migration. The changes of cell cycle and apoptosis were detected by flow cytometry and TUNEL assay. Finally, the effects of Erianin on cell function and signaling pathway-related protein expression in vivo and in vitro were examined based on the enrichment analysis.ResultsErianin could inhibit the proliferation, invasion and migration, induce apoptosis, altered cell cycle of DDP-resistant LUAD cells, and reverse the resistance to DDP. Western blotting results showed that Erianin exerted its anti-tumor effects by regulating the Wnt/β-catenin cascade in DDP-resistant LUAD cells.ConclusionErianin may exerted its anti-tumor effect in DDP-resistant LUAD cells by regulating the Wnt3/β-Catenin/Survivin/Bcl-2/Caspase-3/Cyclin D1 axis.

Rational Design of Self-Reporting Polymersomes for the Controlled Release of Sulfur Dioxide.

As a new member of the gaseous regulators, sulfur dioxide (SO2) plays a crucial role in many biological activities. Recent studies have shown that SO2 is capable of inducing cancer cell apoptosis by regulating intracellular reactive oxygen species (ROS), allowing SO2 to serve as an efficient therapeutic agent. Although various polymer-based platforms have presented great potential for the controlled release of SO2, most of the systems are incapable of monitoring the intracellular generation of SO2. In this work we present the rational design of SO2-releasing biodegradable polymersomes, accompanied by a self-reporting property. The polymersome consists of a hydrophilic block of poly(ethylene glycol) (PEG) and a hydrophobic segment of poly(trimethylene carbonate) (PTMC)-based SO2 donors. The polymersomes not only exhibit good SO2-releasing performance upon treatment with glutathione (GSH), but can also regulate the fluorescence change of the system, offering a good platform for real-time monitoring of the intracellular production of SO2. Significantly, the in vitro and in vivo studies indicate the potential for exploitation of these polymersomes as antitumor agents. We expect that incorporating both the SO2-releasing capacity and self-reporting feature within a polymersome system will provide a unique opportunity for the development of intelligent gas nanovehicles.

Honokiol-loaded PLGA-PEG nanoparticles with solubility in water for infusion treatment of solid cancer

This study aimed to investigate honokiol-loaded PLGA-PEG nanoparticles with good stability in water for infusion treatment. The characteristics of this nanoparticle-based delivery system were analyzed in a previous study. Additionally, fourier transform infrared spectroscopy confirmed the successful loading of honokiol into PLGA-PEG nanoparticles, revealing characteristic chemical shifts. In vitro drug release studies demonstrated sustained release over 72 h, with lower release rates compared to free honokiol. Cytotoxicity assays on A549 and 3LL lung cancer cells showed that the IC50 values for the nanoparticles were significantly lower than those for free honokiol, with values of 8.76 ± 1.51 μg/ml and 7.46 ± 0.53 μg/ml for the nanoparticles, compared to 12.03 ± 1.07 μg/ml and 10.51 ± 1.27 μg/ml for free honokiol, respectively. Apoptosis induction in cancer cells was demonstrated, emphasizing the potential therapeutic benefits. In vivo studies using a murine carcinoma model showed higher concentrations of honokiol in tumor tissues and major organs with the nanoparticle formulation. Tumor growth inhibition was significantly greater in the nanoparticle-treated group, achieving an inhibition ratio (IR) of 85.32 %, compared to 65.99 % for free honokiol. Hematological and biochemical analyses and histopathological examination indicated no significant adverse effects. The study suggests that honokiol-loaded PLGA-PEG nanoparticles covered by carboxyl (COOH) groups (Nano-HK), with good solubility in water, enhance the delivery and therapeutic efficacy of honokiol, providing a promising avenue for cancer treatment.

Unveiling the therapeutic potential of anthocyanin/cisplatin-loaded chitosan nanoparticles against breast and liver cancers

BackgroundLiver and breast cancers are among the leading causes of cancer-related deaths worldwide, prompting researchers to seek natural anticancer agents and reduce chemotherapy side effects. Red beetroot (Beta vulgaris Linnaeus), rich in polyphenols and powerful antioxidants, has shown potential in cancer prevention. This study aimed to evaluate the impact of red beetroot-derived anthocyanin (Ant), Ant-loaded chitosan nanoparticles (Ant NPs), cisplatin (Cis), Cis-loaded chitosan (Cis NPs), and Cis + Ant-loaded chitosan NPs on human hepatoma HepG2 and breast adenocarcinoma MCF7 cell lines.MethodsNPs preparation was evaluated by zeta potential, FTIR, and SEM. The cytotoxic, apoptotic, antioxidant, anti-inflammatory, anti-metastatic, and anti-angiogenic effects were assessed by MTT assay, qPCR, AO/EB staining, and flow cytometry.ResultsTreatment with Ant, Ant NPs, Cis, Cis NPs, and Cis + Ant NPs caused cytotoxicity in HepG2 and MCF7 with best effect in Cis-treated cells. The anticancer effects were attributed to mitochondrial-dependent apoptosis (with high Bax and low Bcl2 expression), chromatin disintegration, and cell cycle arrest in G2/M and S phases. All treatments inhibited migration by downregulating the migration-related gene MMP9 and upregulating the anti-migratory gene TIMP1 and decreased the angiogenesis-related gene VEGF and the inflammatory gene TNFα with best results in Cis NPs-treated cells. Interestingly, Ant, Ant NPs, and Cis + Ant NPs increased the antioxidant status (high GSH and upregulated expression of Nrf2 and OH-1) and decreased drug resistance-related MAPK1 and MDR1 genes compared to Cis and Cis NPs-treated cells.ConclusionsAnthocyanin and cisplatin-loaded chitosan nanoparticles effectively combat breast and liver cancers by inducing cancer cell apoptosis, enhancing antioxidant defenses, and reducing inflammation. They also inhibit tumor spread and blood vessel formation through downregulation of MMP9 and VEGF, highlighting their therapeutic potential.

EXTH-61. TOWARDS A QUANTUM THERAPY FOR GLIOBLASTOMA: A NEW THERAPEUTIC PARADIGM IN MEDICINE

Abstract BACKGROUND A cell operates as an interconnected bioelectrical circuit, utilising electron transfer processes for intracellular communication, with cytochrome c (Cyt c) playing a pivotal role. The redox processes of Cyt c, occurring via electron tunnelling, are essential for its translocation into the cytosol and modulation of its conformation to bind apoptotic protease activating factor 1. This highlights the need for novel technologies capable of interacting with these processes at the atomic scale to control downstream effects and induce apoptosis in cancer cells. METHODS We demonstrate that ‘bio-nanoantennae’, when supplied with an electrical current, enable quantum biological tunnelling for electron transfer (QBET) and facilitate cellular apoptosis in patient-derived IDH wild-type glioblastoma from both the infiltrative tumour margin and proliferative core. The bio-nanoantennae were constructed from gold nanoparticles functionalised with reduced Cyt c and zinc porphyrin as a redox couple. RESULTS Electrical polarisation of these bio-nanoantennae via resonant alternating currents in preclinical glioblastoma cells led to decreased metabolic activity and reduced cell viability by oxidizing Cyt c, thus inducing cellular stress. No significant effect was observed in healthy human astrocyte counterparts. The cytosol localised bio-nanoantennae induced differential gene expression related to ion channels, apoptosis, cancer proliferation and tumour suppression upon activation, in tumour relative to astrocyte cell populations. The bio-nanoantennae were also tested in 3D glioblastoma spheroid models, showing similar effects, and in vivo studies demonstrated a significant reduction in glioblastoma xenograft tumour size. CONCLUSION We propose that bio-nanoantennae modulate the redox state of Cyt c under an electrical field through QBET. To validate this, we investigated the tunnel junction energy and plasmon resonance scattering, and developed a mathematical model to explain the system’s behaviour. This innovative wireless electrical–molecular nanodevice, capable of inducing cancer cell apoptosis, paves the way for further applications of quantum signalling as a new (non-pharmacological) therapeutic paradigm.

Relationship between Apoptosis Induction in Cancer Cells and the Amount of Electrical Stimulation on Frequency Analysis of Nanosecond Pulsed Electric Field

Relationship between Apoptosis Induction in Cancer Cells and the Amount of Electrical Stimulation on Frequency Analysis of Nanosecond Pulsed Electric Field

Lysosome-Targeted Bifunctional Fluorescent Probe and Type I/II Photosensitizer for Viscosity Imaging and Cancer Photodynamic Therapy.

Abnormal lysosomal viscosity is closely associated with cancer progression, underscoring the need for bifunctional fluorescent probes and photosensitizers (PSs) that can both monitor viscosity and facilitate imaging-guided therapy for simultaneous cancer diagnosis and treatment. Despite advances in lysosome-targeted PSs development, few have demonstrated the ability to generate both Type I and Type II reactive oxygen species (ROS). In this study, we present BTTPA, a lysosome-targeted fluorescent probe and photosensitizer, designed to integrate cancer diagnosis via viscosity imaging and cancer treatment through photodynamic therapy (PDT). Our findings reveal that BTTPA selectively targets lysosomes, enabling dynamic monitoring of cellular viscosity and distinguishing cancer cells from normal cells. Upon light activation, BTTPA efficiently generates both Type I and Type II ROS. Apoptosis assays further confirm BTTPA's effectiveness in inducing cancer cell apoptosis, highlighting its potential as a powerful tool for cancer diagnosis and therapy.

Plasma activated solution synergistic with exogenous RONS enhances cancer cell apoptosis for improved tumor therapy

Abstract The role of reactive oxygen and nitrogen species (RONS) in plasma-activated solution (PAS) for inducing cancer cell apoptosis is well-established, but suboptimal concentrations and rapid degradation often limit their efficacy. This study addresses a significant gap in current research by exploring how the incorporation of exogenous RONS into PAS can enhance apoptosis in cancer cells, potentially overcoming the limitations of traditional PAS treatments. In this study, the effect of treating saline with a plasma jet for different durations was investigated to analyze how its chemical composition triggers apoptosis in cancer cells in vitro. It was observed that longer plasma treatment times resulted in lower cell viability, the reactive species of H2O2, NO2- and ONOO- were found to play a crucial role in inducing apoptosis in cancer cells. Based on these findings, the anticancer efficacy of PAS was further evaluated after the addition of specific exogenous RONS, using measures such as cell viability, intracellular ROS levels, and microscopic imaging. Biological experiments demonstrated that the anticancer efficacy followed the order: addition of RONS to saline before plasma discharge treatment > addition of RONS to saline after plasma discharge treatment > exogenous RONS solution without plasma treatment. This suggests that the external introduction of RONS can modify the chemical composition of PAS to effectively enhance its anticancer properties, with the addition of H2O2 before plasma discharge treatment proving to be the most effective strategy. These results provide a novel perspective on harnessing the anticancer potential of PAS and expanding its potential applications in tumor therapy.