毒胡萝卜素对胃癌BCG-823细胞凋亡的影响  

Effect of thapsigarg on apoptosis of BCG-823 cells in gastric cancer

作　　者：陈玲 吕小婷 孙蕾清 郑璐 周忠海 陈复兴 李洪春 CHEN Ling;LV Xiao-ting;SUN Lei-qing;ZHENG Lu;ZHOU Zhong-hai;CHEN Fu-xing;LI Hong-chun(College of Medical Technology,Xuzhou Medical University,Xuzhou 221004,Jiangsu,China;Department of Central Laboratory,the 71th Hospital of Army,Xuzhou 221004,Jiangsu,China;Department of Clinical Laboratory,the Affiliated Hospital of Xuzhou Medical University,Xuzhou 221002,Jiangsu,China)

机构地区：[1]徐州医科大学医学技术学院,江苏徐州221004 [2]陆军第七十一集团军医院实验科,江苏徐州221004 [3]徐州医科大学附属医院检验科,江苏徐州221002

出　　处：《生物医学工程与临床》2021年第2期219-224,共6页Biomedical Engineering and Clinical Medicine

基　　金：南京军区医学科技创新重点课题(14ZD17)。

摘　　要：目的探讨毒胡萝卜素对胃癌BCG-823细胞凋亡的影响及机制研究。方法体外培养BCG-823细胞,分别经不同浓度的毒胡萝卜素(0.00、0.75、1.50、3.00、6.00、12.00μmol/L)诱导培养24 h和48 h。采用CCK-8法检测毒胡萝卜素对BCG-823细胞活力的影响和计算生长抑制率;流式细胞术检测胃癌细胞的凋亡和周期情况;Western blot检测Bcl-2、Bax、Cyclin D1、Cyt C、cleaved-Caspase 9、cleaved-Caspase 3蛋白表达水平。结果毒胡萝卜素诱导胃癌BCG-823细胞24 h后,BCG-823细胞的活力受到明显抑制作用,并具有浓度依赖性。随着毒胡萝卜素作用浓度的增加,BCG-823细胞的抑制率随之增加,由(0.23±0.04)%增加到(47.23±1.85)%。随着浓度的增加G1/G0期比例显著增加,S期和G_(2)/M期细胞比例依次降低,即浓度0.00μmol/L时G1/G0期为(35.92±2.35)%,S期(62.88±2.36)%,G_(2)期(1.20±0.02)%;浓度为12.00μmol/L时G1/G0期为(88.98±1.86)%,S期(9.84±0.86)%,G_(2)期(1.18±0.02)%。毒胡萝卜素对BCG-823细胞的细胞凋亡率具有浓度依赖性,在浓度12.00μmol/L时BCG-823细胞的细胞凋亡率(38.95%±2.43%)最高,与浓度0.00μmol/L时(4.90%±1.33%)比较,差异有显著统计学意义(P<0.01)。随着毒胡萝卜素浓度的增加促凋亡蛋白Bax和Cyt C表达依次增加,抑凋亡蛋白Bcl-2和周期蛋白Cyclin D1表达随之减少,凋亡相关蛋白cleaved-Caspase 9、cleaved-Caspase 3表达量随之增加。结论毒胡萝卜素可促进胃癌BCG-823细胞凋亡,通过阻滞细胞周期在G1期和启动线粒体凋亡途径相关。Objective To investigate the effect of thapsigarg on apoptosis of BCG-823 cells in gastric cancer and its mechanism.Methods BCG-823 cells were cultured in vitro and cultured with different concentrations of thapsigarg(0.00,0.75,1.50,3.00,6.00,12.00μmol/L)for 24 hours and 48 hours.CCK-8 method was used to detect the effect of thapsigarg on activity of BCG-823 cells and calculate growth inhibition rate.Flow cytometry was used to detect the apoptosis and cycle of gas-tric cancer cells.Western blot was used to detect protein expression levels of Bcl-2,Bax,Cyclin D1,Cyt C,cleaved-Caspase 9 and cleaved-Caspase 3.Results The activity of BCG-823 cells was significantly inhibited after 24-hour induction by thapsigarg,and the inhibitation was concentration dependent.With increased concentration of thapsigarg,the apoptosis rate of BCG-823 cells increased from(0.23±0.04)%to(47.23±1.85)%;the G1/G0 ratio increased significantly and proportions of S stage and G_(2)/M stage cells decreased successively.With increased concentration,the cell ratio in G1/G0 stage increased significantly,and decreased sequentially in S stage and G_(2)/M stage,which were(35.92±2.35)%in G1/G0 stage,(62.88±2.36)%in S stage and(1.20±0.02)%in G_(2)stage at concentration of 0.00μmol/L;(88.98±1.86)%in G1/G0 stage,(9.84±0.86)%in S stage and(1.18±0.02)%in G_(2)stage at concentration of 12.00μmol/L.Thalassene was concentration-dependent on apoptosis rate of BCG-823 cells,and at concentration of 12.00μmol/L,the apoptosis rate of BCG-823 cells(38.95%±2.43%)was the highest,compared with that at concentration of 0.00μmol/L(4.90%±1.33%),the difference was statistically significant(P<0.01).With increased thapsigargin concentration,the expression of pro-apoptotic proteins Bax and Cyt C increased,the expression of anti-apoptotic proteins Bcl-2 and Cyclin D1 decreased,and expression of apoptosis-related proteins cleaved-Caspase 9 and cleaved-Caspase 3 increased.Conclusion It is demonstrated that thapsigarg can promote apoptosis of gastric cancer BCG-823

关 键 词：毒胡萝卜素 BCG-823 细胞凋亡 细胞周期 线粒体通路 

分 类 号：R735.2[医药卫生—肿瘤]