正得康诱导肺癌细胞凋亡及抑制肺癌转移的线粒体通路机制OA
Mechanism of Zhengdekang in Inducing Apoptosis and Inhibiting Metastasis of Lung Cancer Cells via the Mitochondrial Pathway
目的 探究中药复方正得康抑制肺癌转移的作用及机制.方法 体外采用CCK-8 实验、平板克隆形成实验、Transwell细胞迁移实验和流式细胞凋亡实验评估正得康对人肺腺癌循环肿瘤细胞(CTC-TJH-01)和小鼠Lewis肺癌(LLC)细胞增殖、迁移和凋亡的影响;通过JC-10 线粒体膜电位染色实验评估正得康对CTC-TJH-01和LLC细胞线粒体膜电位的影响;采用Western Blot实验检测经正得康干预后肺癌细胞内线粒体/细胞色素C通路相关蛋白Bcl-2、Bax、Cytochrome C、Caspase-9 和Cleaved-Caspase-3 的表达水平.体内采用LLC细胞在C57BL/6J小鼠上构建尾静脉注射肺转移模型,给予正得康每日 1 次灌胃给药,25 d后取材,拍照观察小鼠的肺癌转移情况;HE染色观察肺组织病理变化;免疫组化法检测肺转移瘤组织中Ki-67 和Cleaved-Caspase-3蛋白的表达水平.结果 正得康可明显抑制CTC-TJH-01 和LLC细胞的增殖和迁移(P＜0.05,P＜0.01,P＜0.001),并明显诱导肺癌细胞发生凋亡(P＜0.001);随着正得康给药浓度的增加,CTC-TJH-01 和LLC细胞内线粒体膜电位逐渐下降(P＜0.01,P＜0.001);正得康干预后还可下调CTC-TJH-01 和LLC细胞中Bcl-2 蛋白的表达水平(P＜0.05),上调Bax、Cytochrome C、Caspase-9 和Cleaved-Caspase-3 蛋白的表达水平(P＜0.05,P＜0.01,P＜0.001).体内研究发现正得康给药后可明显降低小鼠肺转移瘤数目(P＜0.01),并且下调肺转移瘤组织中Ki-67 蛋白的表达水平(P＜0.001),上调Cleaved-Caspase-3 蛋白的表达水平(P＜0.001).结论 正得康具有通过调控内源性线粒体凋亡途径诱导肺癌细胞凋亡,进而抑制肺癌转移的作用.
Objective To investigate the effect and mechanism of the Chinese herbal compound Zhengdekang(ZDK)in inhibiting lung cancer metastasis.Methods In vitro,CCK-8 assay,plate colony formation assay,Transwell migration assay,and flow cytometric apoptosis assay were used to evaluate the effects of ZDK on the proliferation,migration,and apoptosis of human lung adenocarcinoma circulating tumor cells(CTC-TJH-01)and mouse Lewis lung carcinoma(LLC)cells.JC-10 mitochondrial membrane potential staining was employed to assess the effect of ZDK on the mitochondrial membrane potential in CTC-TJH-01 and LLC cells.Western Blot was performed to detect the expression levels of mitochondrial/cytochrome C pathway-related proteins,including Bcl-2,Bax,Cytochrome C,Caspase-9,and Cleaved-Caspase-3,in lung cancer cells after ZDK intervention.In vivo,a lung metastasis model was established in C57BL/6J mice via tail vein injection of LLC cells.The mice were administered ZDK by gavage once daily for 25 days.Lung metastases were observed and photographed after sample collection.Pathological changes in lung tissue were examined by HE staining,and the expression levels of Ki-67 and Cleaved-Caspase-3 proteins in lung metastatic tissues were detected by immunohistochemistry.Results ZDK significantly inhibited the proliferation and migration of CTC-TJH-01 and LLC cells(P＜0.05,P＜0.01,P＜0.001)and markedly induced apoptosis in lung cancer cells(P＜0.001).With increasing concentrations of ZDK,the mitochondrial membrane potential in CTC-TJH-01 and LLC cells gradually decreased(P＜0.01,P＜0.001).ZDK intervention also downregulated the expression of Bcl-2 protein(P＜0.05)and upregulated the expression of Bax,Cytochrome C,Caspase-9,and Cleaved-Caspase-3 proteins in CTC-TJH-01 and LLC cells(P＜0.05,P＜0.01,P＜0.001).In vivo studies showed that ZDK administration significantly reduced the number of lung metastatic nodules in mice(P＜0.01),downregulated the expression of Ki-67 protein(P＜0.001),and upregulated the expression of Cleaved-Caspase-3 protein in lung metastatic tissues(P＜0.001).Conclusion ZDK inhibits lung cancer metastasis by inducing apoptosis in lung cancer cells through regulation of the endogenous mitochondrial apoptotic pathway.
刘诗卉;李佳轩;田建辉;刘佳君;张志鹏;阙祖俊
上海中医药大学附属市中医医院肿瘤临床医学中心,上海 200071上海中医药大学附属市中医医院肿瘤研究所,上海 200071上海中医药大学附属市中医医院肿瘤临床医学中心,上海 200071上海中医药大学附属市中医医院肿瘤研究所,上海 200071上海中医药大学附属市中医医院肿瘤临床医学中心,上海 200071上海中医药大学附属市中医医院肿瘤研究所,上海 200071
医药卫生
正得康肺癌细胞凋亡线粒体转移人肺腺癌循环肿瘤细胞小鼠Lewis肺癌细胞小鼠
Zhengdekanglung cancerapoptosismitochondriametastasishuman lung adenocarcinoma circulating tumor cellsmouse Lewis lung carcinoma cellsmice
《中药新药与临床药理》 2026 (1)
42-51,10
国家自然科学基金面上项目(82174245)上海市炎癌转化病证生物学前沿研究基地(2021KJ03-12)上海市卫生健康领军人才(2022LJ014).
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