槐果碱对人肠癌细胞HCT-116生长抑制作用  

Effect of sophocarpine on the growth of human HCT 116 colon cancer cells

作　　者：颜勋 刘成宸 张明[2] 陆峰[2] 周广军 YAN Xun;LIU Chengchen;ZHANG Ming;LU Feng;ZHOU Guangjun(Yancheng Clinical College,Xuzhou Medical University,Yancheng,Jiangsu 224000,China;Department of General Surgery,the People′s Hospital of Binhai County,Binhai,Jiangsu 224000)

机构地区：[1]徐州医科大学附属盐城临床医学院,江苏盐城224000 [2]滨海县人民医院普外科,江苏滨海224000

出　　处：《徐州医科大学学报》2023年第5期367-372,共6页Journal of Xuzhou Medical University

基　　金：滨海县人民医院科研项目(BHKY201801);江苏大学临床医学科技发展基金(JLY20180198)。

摘　　要：目的探讨槐果碱对人肠癌细胞HCT-116生长的影响。方法选取人肠癌细胞HCT-116,用不同浓度梯度槐果碱作用48 h后,通过细胞毒性试验检测增殖能力,根据IC_(50)选择3个浓度进一步实验。通过平板克隆形成实验检测细胞克隆能力,流式细胞术检测细胞凋亡情况,线粒体膜电位检测线粒体膜电位水平,蛋白质印迹法检测凋亡相关蛋白及生存素(survivin)的表达水平。结果细胞毒性试验显示,槐果碱可抑制HCT-116肿瘤细胞增殖,且随着槐果碱浓度的升高抑制作用增强。IC_(50)为2.134 mmol/L,并选择1、2和4 mmol/L进行进一步相关实验;在槐果碱作用后HCT-116细胞克隆能力、线粒体膜电位水平均明显下降,细胞凋亡率明显升高(P<0.05);槐果碱作用后,HCT-116细胞内Bcl-xL、Bcl-2、survivin表达显著下调,而Bax、Cleaved Caspase-3和Cleaved Caspase-9显著上调。结论槐果碱可抑制肠癌细胞HCT-116增殖并通过线粒体途径促进凋亡。Objective To investigate the effect of sophocarpine on the growth of human HCT 116 colon cancer cells.Methods HCT-116 cells were treated with sophocarpine at different concentrations for 48 h.Their proliferation was detected by the cytotoxic assay.According to IC_(50) values,three concentrations were selected for further experiments.Then,the capability of a single cell to form a colon was measured by the clonogenic assay.The apoptosis was evaluated by flow cytometry.The mitochondrial membrane potential were determined by the mitochondrial membrane potential assay.The levels of apoptosis-associated proteins and survivin were measured by Western blot.Results According to the cytotoxic assay,sophocarpine inhibited the proliferation of HCT-116 cells and the inhibitory effect increased with the rising of sophocarpine concentrations.The IC_(50) value was 2.134 mmol/L and then 1,2 and 4 mmol/L sophocarpine were selected for further experiments.After treatment with sophocarpine,remarkable decreases were see in the clonogenic capability and mitochondrial membrane potentials,while apoptosis was enhanced(P<0.05).Furthermore,treatment with sophocarpine resulted in downregulated expression of Bcl-xL,Bcl-2 and survivin in HCT-116 cells after,and upregulated expression of Bax,Cleaved Caspase-3,and Cleaved Caspase-9.Conclusions Sophocarpine inhibits the proliferation of HCT-116 cells and promoted apoptosis via the mitochondrial pathway.

关 键 词：槐果碱 肠癌细胞HCT-116 线粒体途径 凋亡 SURVIVIN 

分 类 号：R73-361[医药卫生—肿瘤]