The Chinese Herbal Formula Weichang’an Reduces the Proliferation of Human Gastric Cancer Cells via Mitochondrial Apoptosis  

中药方剂胃肠安通过线粒体凋亡减少人胃癌细胞的增殖

作　　者：Weixia Chen Yaofei Niu Bin Chen Aiguang Zhao 陈伟霞;牛篧飞;陈彬;赵爱光(河南省中医院,河南郑州;上海中医药大学龙华医院,上海;河南省人民医院/郑州大学人民医院,河南郑州;上海中医药大学普陀医院,上海)

机构地区：[1]Department of Oncology,Henan Province Hospital of Traditional Chinese Medicine,Zhengzhou,Henan,China [2]Department of Oncology,Longhua Hospital Shanghai University of Traditional Chinese Medicine,Shanghai,China [3]Department of Infection,Henan Provincial People’s Hospital/Zhengzhou University People’s Hospital,Zhengzhou,Henan,China [4]Department of Oncology,Putuo Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai,China

出　　处：《Chinese Medicine and Natural Products》2024年第3期117-125,共9页中医学报（英文）

基　　金：funded by the Three-Year Plan of Action for the Development of Traditional Chinese Medicine in Shanghai(ZY3-CCCX-3-2003);State Administration of Traditional Chinese Medicine(TCM)Clinical Research Base of China(JDZX2015068);the Key and Promotion Special Topics of Henan(202102310164);Henan Province Traditional Chinese Medicine Scientific Research Special Project(2022ZY2031).

摘　　要：Objective The aim of the study was to investigate the effects of Chinese herbal formula Weichang’an(WCA)on the proliferation and mitochondria-mediated apoptosis of human gastric cancer cells.Methods Cell Counting Kit-8(CCK8)was used to evaluate the antiproliferative activity ofWCA on MKN45 cells;Giemsa staining was used to investigate cell colony formation;flow cytometry was used to analyze cell cycle,apoptosis rate,and caspases activation;and Hoechst staining was used to analyze the morphology of cell nuclei.The mitochondrial membrane potential was analyzed by both flow cytometric measurements and fluorescence microscopy.Western blot was used to analyze the protein levels of pro-caspase-3,Bcl-2,Bax,and Bcl-X.MKN45 cells were subcutaneously injected into the right forelimb of 16 nude mice to establish the tumor xenograft model,and a total of 12 nude mouse tumor xenograft models were successfully created.The nude mice were divided into the control group and the WCA-treated group.The two groups received normal saline and WCA treatment at 35.49 g/kg by a single daily oral gavage for 21 days.The body weight and tumor size of the nude mice were measured twice a week.The ultrastructural changes of subcutaneous tumors were observed by a transmission electron microscope.Results WCA can suppress cell proliferation and colony formation.It can also induce changes in the mitochondrial membrane potential and increase the activities of caspases-3,caspases-8,and caspases-9 in MKN45 cells.WCA induced S-phase arrest and apoptosis in MKN45 cells,and decreased the expression levels of antiapoptotic proteins Bcl-2 and Bcl-X in MKN45 cells,while increasing the expression levels of the proapoptotic proteins Bax and pro-caspase-3 compared with the control group.WCA inhibited the growth of a xenografted MKN45 tumor in nude mice,and the protein levels of Bax and pro-caspase-3 were significantly increased,while Bcl-X and Bcl-2 were reduced compared with the control group.The differences are statistically significant(p<0.05).Conclusion W目的:研究中药方剂胃肠安(Weichang′an,WCA)对人胃癌细胞增殖及线粒体介导的凋亡的影响。方法:使用CCK8评估胃肠安对MKN45细胞的抗增殖活性;利用吉姆萨染色观察细胞集落形成;流式细胞术分析细胞周期、凋亡率和Caspases活性;利用Hoechst染色分析细胞核形态;通过流式细胞术和荧光显微镜分析线粒体膜电位;利用蛋白质印迹法分析pro-Caspase-3、Bcl-2、Bax和Bcl-X的蛋白水平。将MKN45细胞皮下注射到16只裸鼠右前肢建立肿瘤异种移植模型,共成功建立了12只裸鼠肿瘤异种移植模型。裸鼠分为对照组和胃肠安处理组,两组分别接受生理盐水和胃肠安处理,剂量为35.49 g·kg^(-1),每日1次口服灌胃,持续21天。每周两次测量裸鼠的体质量和肿瘤大小。通过透射电子显微镜观察皮下肿瘤的超微结构变化。结果:胃肠安可抑制细胞增殖、集落形成,并改变了线粒体膜电位,提高MKN45细胞中Caspase-3、Caspase-8和Caspase-9的活性;与对照组相比,胃肠安诱导MKN45细胞S期阻滞和凋亡,减少MKN45细胞中抗凋亡蛋白Bcl-2和Bcl-X的表达,同时增加了促凋亡蛋白Bax和pro-Caspase-3的表达;与对照组相比,胃肠安抑制了裸鼠异种移植MKN45肿瘤的生长,Bax和pro-Caspase-3的蛋白水平显著升高,而Bcl-X和Bcl-2的水平降低,且差异具有统计学意义(P<0.05)。结论:胃肠安可通过线粒体凋亡抑制胃癌细胞的增殖。

关 键 词：gastric cancer Chinese herbal formula Weichang’an mitochondrial apoptosis 

分 类 号：R273[医药卫生—中西医结合]