Phytomedicine : international journal of phytotherapy and phytopharmacology · 2026
Zuojin Wan reverses cetuximab resistance in KRAS-mutant colorectal cancer via the NRF2-GCH1/CoQ and DHODH/CoQ axes.
СтатьяНазвание и аннотация: Перевод пока не подготовлен
Обработка статьи · этапы, ошибки и управление
Загрузка состояния…
Ошибки и ограничения
История · было → стало
Состояние обновляется каждые 5 секунд, пока блок открыт. Процент показывается только там, где известен объём работы. Пауза останавливает продолжение этапа; текущий запрос к источнику может завершиться.
Liu Xiaodie, Chen Yongqi, Jiang Yulang, Sui Hua, Liang Tingyu, Meng Jiao, Chang Bin, Ji Qing, Wang Ziyuan
Аннотация
BACKGROUND: Cetuximab (CET), a monoclonal antibody targeting the epidermal growth factor receptor (EGFR), serves as a cornerstone therapeutic option for advanced colorectal cancer (CRC). However, patients with KRAS-mutant CRC present inherent resistance to CET, which directly compromises its clinical efficacy and therapeutic benefits. Zuojin Wan (ZJW), a classic traditional Chinese medicine formula consisting of Coptis chinensis Franch. and Evodia rutaecarpa (Juss.) Benth., has been documented to exert potent antitumor activity; nevertheless, its potential role in reversing CET resistance in KRAS-mutated CRC and the underlying molecular mechanisms remain poorly defined.
PURPOSE: This study aimed to investigate the efficacy of ZJW in overcoming CET resistance in KRAS-mutated CRC and further characterize its key molecular targets and regulatory mechanisms.
METHODS: In vitro experiments were conducted using two KRAS-mutated CRC cell lines, HCT116 and SW620. The synergistic antiproliferative effect of ZJW combined with CET was assessed via CCK-8 assay and colony formation assay. Ferroptosis induction mediated by the combination treatment was verified by measuring lipid peroxidation, glutathione (GSH) depletion, ferrous ion (Fe2+) accumulation, and observing characteristic mitochondrial morphological alterations using transmission electron microscopy (TEM). Reactive oxygen species (ROS) levels were detected to evaluate intracellular oxidative stress. Transcriptome sequencing, Western blot analysis and enzyme-linked immunosorbent assay (ELISA) were performed to explore the underlying molecular mechanisms, while plasmid transfection and quantitative real-time polymerase chain reaction (qRT-PCR) were carried out for functional validation. A nude mouse subcutaneous xenograft model was established to evaluate the antitumor efficacy of the combined treatment in vivo, and immunohistochemical (IHC) staining was performed to detect the expression of relevant proteins. The primary bioactive components of ZJW were identified by mass spectrometry, and network pharmacology analysis was performed to predict the potential interactions between these active components and CRC-related signaling pathways.
RESULTS: ZJW synergistically augmented the CET-mediated suppression of KRAS-mutated CRC cell proliferation and triggered significant ferroptosis, as evidenced by elevated lipid peroxidation, reduced GSH levels, increased Fe2+ accumulation, shrunken mitochondria with condensed cristae, and markedly enhanced intracellular oxidative stress. Combination treatment with ZJW and CET significantly downregulated the expression of core proteins involved in the parallel NRF2-GCH1/CoQ and DHODH/CoQ antioxidant axes, thereby disrupting intracellular redox homeostasis. Silencing of NRF2 or DHODH facilitated ferroptosis and sensitized CRC cells to CET, whereas overexpression of these two molecules abolished the synergistic antitumor effect of ZJW plus CET. In the nude mouse xenograft model, the combined regimen notably suppressed tumor growth and decreased tumor weight, and modulated the expression of ferroptosis-associated markers (NRF2, DHODH) and the lipid peroxidation indicator 4-HNE in tumor tissues. Mass spectrometry identified the major bioactive constituents of ZJW, and network pharmacology analysis predicted their potential involvement in multiple CRC-related pathways, including antioxidant defense and ferroptosis regulation.
CONCLUSION: ZJW reverses CET resistance in KRAS-mutated CRC by inhibiting the parallel NRF2-GCH1/CoQ and DHODH/CoQ antioxidant axes to induce ferroptosis. This study provides a solid scientific basis for the clinical translation of ZJW as an adjuvant therapeutic strategy to boost the efficacy of CET in CRC treatment.
PURPOSE: This study aimed to investigate the efficacy of ZJW in overcoming CET resistance in KRAS-mutated CRC and further characterize its key molecular targets and regulatory mechanisms.
METHODS: In vitro experiments were conducted using two KRAS-mutated CRC cell lines, HCT116 and SW620. The synergistic antiproliferative effect of ZJW combined with CET was assessed via CCK-8 assay and colony formation assay. Ferroptosis induction mediated by the combination treatment was verified by measuring lipid peroxidation, glutathione (GSH) depletion, ferrous ion (Fe2+) accumulation, and observing characteristic mitochondrial morphological alterations using transmission electron microscopy (TEM). Reactive oxygen species (ROS) levels were detected to evaluate intracellular oxidative stress. Transcriptome sequencing, Western blot analysis and enzyme-linked immunosorbent assay (ELISA) were performed to explore the underlying molecular mechanisms, while plasmid transfection and quantitative real-time polymerase chain reaction (qRT-PCR) were carried out for functional validation. A nude mouse subcutaneous xenograft model was established to evaluate the antitumor efficacy of the combined treatment in vivo, and immunohistochemical (IHC) staining was performed to detect the expression of relevant proteins. The primary bioactive components of ZJW were identified by mass spectrometry, and network pharmacology analysis was performed to predict the potential interactions between these active components and CRC-related signaling pathways.
RESULTS: ZJW synergistically augmented the CET-mediated suppression of KRAS-mutated CRC cell proliferation and triggered significant ferroptosis, as evidenced by elevated lipid peroxidation, reduced GSH levels, increased Fe2+ accumulation, shrunken mitochondria with condensed cristae, and markedly enhanced intracellular oxidative stress. Combination treatment with ZJW and CET significantly downregulated the expression of core proteins involved in the parallel NRF2-GCH1/CoQ and DHODH/CoQ antioxidant axes, thereby disrupting intracellular redox homeostasis. Silencing of NRF2 or DHODH facilitated ferroptosis and sensitized CRC cells to CET, whereas overexpression of these two molecules abolished the synergistic antitumor effect of ZJW plus CET. In the nude mouse xenograft model, the combined regimen notably suppressed tumor growth and decreased tumor weight, and modulated the expression of ferroptosis-associated markers (NRF2, DHODH) and the lipid peroxidation indicator 4-HNE in tumor tissues. Mass spectrometry identified the major bioactive constituents of ZJW, and network pharmacology analysis predicted their potential involvement in multiple CRC-related pathways, including antioxidant defense and ferroptosis regulation.
CONCLUSION: ZJW reverses CET resistance in KRAS-mutated CRC by inhibiting the parallel NRF2-GCH1/CoQ and DHODH/CoQ antioxidant axes to induce ferroptosis. This study provides a solid scientific basis for the clinical translation of ZJW as an adjuvant therapeutic strategy to boost the efficacy of CET in CRC treatment.