川楝素诱导人胃癌MGC-803细胞凋亡及其机制  

Toosendanin induces apoptosis of human gastric cancer MGC-803 cells and its mechanism

作　　者：徐兴军[1,2] 李珊珊 刘畅[1] 张伟伟[1,2] 张珍珠[1,2] 邵淑丽 XU Xing-jun;LI Shan-shan;LIU Chang;ZHANG Wei-wei;ZHANG Zhen-zhu;SHAO Shu-li(College of Life Science and Agriculture and Forestry,Qiqihar University,Qiqihaer 161006;Heilongjiang Provincial Key Laboratory of Resistance Gene Engineering and Protection of Biodiversity in Cold Areas,Qiqihar University,Qiqihaer 161006,China)

机构地区：[1]齐齐哈尔大学生命科学与农林学院,黑龙江齐齐哈尔161006 [2]抗性基因工程与寒地生物多样性保护黑龙江省重点实验室,黑龙江齐齐哈尔161006

出　　处：《中国应用生理学杂志》2021年第3期262-265,292,共5页Chinese Journal of Applied Physiology

基　　金：齐齐哈尔大学黑龙江省教育厅基本业务专项重点项目(135109104);黑龙江省高教强省优势特色学科—玉米“粮头食尾”重点专项(LTSW201737);黑龙江省省属高等学校基本科研业务费科研项目(植物性食品加工技术特色学科专项)(YSTSXK201809)。

摘　　要：目的:本研究旨在探讨川楝素诱导人胃癌MGC-803细胞凋亡及其机制。方法:将人胃癌MGC-803细胞分为5组,每组3个复孔,采用氟尿嘧啶(5-FU)和0 nmol/L川楝素(TSN)分别作为阳性对照和阴性对照。其余3组分别加入终浓度为30 nmol/L、50 nmol/L、70 nmol/L的川楝素。川楝素处理细胞48 h后,利用激光共聚焦显微镜观察细胞形态结构变化;流式细胞术检测线粒体膜电位变化;酶标法检测Caspase-3和Caspase-9活性;利用qRT-PCR和Western blot检测凋亡相关基因Bcl-2、Bax、Cyt c和APAF-1 mRNA和蛋白水平。结果:与0 nmol/L TSN组相比,30 nmol/L、50 nmol/L、70 nmol/L的川楝素作用于人胃癌MGC-803细胞48 h,可见细胞体积缩小,细胞核裂解,部分染色质凝集等形态学变化;Caspase-3和Caspase-9活性升高(P<0.05);而线粒体膜电位明显下降(P<0.05);Bax、Cyt c和APAF-1基因mRNA及蛋白表达量显著升高(P<0.05),Bcl-2基因mRNA及蛋白表达量显著降低(P<0.05)。结论:川楝素通过上调Bax、Cyt c和APAF-1的表达,下调Bcl-2基因表达,增强Caspase-3、Caspase-9活性诱导人胃癌MGC-803细胞凋亡。Objective:To explore the apoptosis of human gastric cancer MGC-803 cells induced by toosendanin(TSN)and its mechanism.Methods:The human gastric cancer MGC-803 cells were divided into 5 groups,each group was set with 3 replicate.Fluorouracil(5-FU)and 0 nmol/L toosendanin(TSN)were used as positive control and negative control,respectively.The other three groups were treated with toosendanin at the final concentrations of 30,50,and 70 nmol/L,respectively.After 48 h of treatment with toosendanin,the morphology of the cells were observed under laser confocal microscopy.Flow cytometry was used to detect the mitochondrial membrane potential,and enzyme-labeled assays were used to detect the activities of Caspase-3 and Caspase-9.The mRNA and protein levels of Bcl-2,Bax,Cyt c and APAF-1 were measured by qRT-PCR and Western blot.Results:Compared with the 0 nmol/L TSN group,after the human gastric cancer MGC-803 cells were treated with toosendanin at the concentrations of 30,50,and 70 nmol/L for 48 h,the cell volume shrinkage,nucleus cleavage and chromatin morphological changes were observed under the microscope.The activities of Caspase-3 and Caspase-9 were increased significantly(P<0.05),while the mitochondrial membrane potential was decreased significantly(P<0.05).In addition,the mRNA and protein expression levels of Bax,Cyt c and APAF-1 were increased significantly(P<0.05),while the mRNA and protein expression levels of Bcl-2 were decreased significantly(P<0.05).Conclusion:Toosendanin up-regulates the expressions of Bax,Cyt c and APAF-1,down-regulates the expression of Bcl-2 gene,enhances the activities of caspase-3 and caspase-9,and induces the apoptosis of human gastric cancer MGC-803 cells.

关 键 词：川楝素 MGC-803细胞 细胞凋亡 

分 类 号：Q255[生物学—细胞生物学]