首页|乙氧基二氢血根碱靶向EGFR抑制结直肠癌细胞恶性生物学行为并诱导凋亡的机制研究

乙氧基二氢血根碱靶向EGFR抑制结直肠癌细胞恶性生物学行为并诱导凋亡的机制研究

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目的 探讨乙氧基二氢血根碱(ethoxydihydrosanguinarine，ESG)对结直肠癌细胞的抑制及具体分子机制。方法 选用结直肠癌细胞系RKO和HRT-18。实验分为空白对照组和加药组(1。0、1。5、2。0 μmol/L ESG)。通过CCK-8法、实时无标记细胞分析技术(real time cellular analysis，RTCA)及克隆形成实验检测ESG对细胞增殖能力的影响;以流式细胞术、DAPI染色、JC-1线粒体膜电位和活性氧检测试剂盒检测ESG对细胞凋亡的影响;通过高内涵成像分析技术及Transwell侵袭实验检测ESG对细胞侵袭、迁移的影响;Western blot检测细胞凋亡及侵袭、迁移相关蛋白的表达，表皮生长因子受体(epidermal growth factor receptor，EGFR)以及下游Akt/ERK信号通路蛋白的表达及磷酸化水平;通过过表达EGFR和敲低EGFR探讨EGFR在ESG诱导结直肠癌细胞凋亡、抑制细胞增殖及侵袭迁移过程中的作用;通过分子对接、靶点稳定性药物亲和反应实验以及微量热泳动实验检测ESG与EGFR的直接结合作用。结果 ESG显著抑制结直肠癌细胞的增殖及克隆形成能力;ESG诱导结直肠癌细胞发生线粒体途径凋亡并抑制其侵袭、迁移能力;ESG直接结合EGFR的激酶活性区而抑制其活性，进而下调其下游与肿瘤恶性进展相关的信号分子Akt、ERK的磷酸化水平。结论 ESG靶向结合并抑制EGFR，进而通过其下游Akt/ERK信号通路抑制结直肠癌细胞侵袭、迁移并诱导线粒体途径的细胞凋亡，有望成为靶向EGFR治疗结直肠癌的新型靶向药物。
The Mechanism of Ethoxydihydrosanguinarine Targeting EGFR to Inhibit Malignant Progression and Induce Apoptosis of Colorectal Cancer
Objective To investigate the antitumor effects of ethoxydihydrosanguinarine(ESG)on colorectal cancer cells and its possible mechanisms.Methods Colorectal cancer cell lines RKO and HRT-18 were used.The cells were divided into blank control group and drug treatment groups(1.0,1.5,2.0 μmol/L ESG).CCK-8 assay,real time cellular analysis(RTCA)and colo-ny formation assay were used to detect the effect of ESG on cell proliferation and survival.Flow cytometry,DAPI staining,JC-1 mitochondrial membrane potential assay,and reactive oxygen species detection were performed to detect the effect of ESG on cell apoptosis.High-content analysis and Transwell invasion assay were used to detect the effect of ESG on cell invasion and mi-gration.Western blot was used to detect the expression of apoptosis,invasion and migration-related proteins,and the expression and phosphorylation of epidermal growth factor receptor(EGFR)and Akt/ERK signaling pathway proteins.The roles of EGFR in ESG-induced apoptosis,proliferation,invasion,and migration inhibition in colorectal cancer cells were detect through overex-pression and knockdown of EGFR.The direct binding between ESG and EGFR was detected by molecular docking and drug af-finity responsive target stability assay.Results The result showed that the proliferation and colony formation ability of colorec-tal cancer cells were significantly inhibited by ESG.Apoptosis through mitochondrial pathway was induced by ESG.The inva-sion and migration ability of colorectal cancer cells were inhibited by ESG.Moreover,ESG directly bound to EGFR and down-regulated the phosphorylation levels of Akt and ERK.Conclusion ESG targets and inhibits EGFR,thereby inhibiting the malig-nant progression of colorectal cancer cells and inducing mitochondrial apoptosis through its downstream Akt/ERK signaling pathway.This study provides new targeted drugs for targeting EGFR in the treatment of colorectal cancer.

colorectal cancerethoxydihydrosanguinarineepidermal growth factor receptorinvasionmigrationapoptosis

谭苗、钱琛、向可、段钟琪、万芳、刘莹

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湖北医药学院 基础医学院,十堰 442000

湖北医药学院 武当特色中药研究湖北省重点实验室,十堰 442000

湖北文理学院附属谷城医院,襄阳 441700

结直肠癌 乙氧基二氢血根碱 表皮生长因子受体 侵袭 迁移 凋亡

国家自然科学基金资助项目湖北省卫生健康委指导性项目湖北医药学院研究生科技创新项目2022年大学生创新创业训练计划项目2023年大学生创新创业训练计划项目

82072928WJ2023F079YC2022032202210929007S202310929001

2024

10.3870/j.issn.1672-0741.23.06.008

华中科技大学学报(医学版)
华中科技大学

华中科技大学学报(医学版)

CSTPCD北大核心
影响因子：1.443
ISSN：1672-0741
年,卷(期)：2024.53(3)