环匹酮胺抑制人肝癌细胞株HepG2增殖和促进细胞凋亡作用的机制研究  

Study on the mechanism of cyclopiroxamine inhibiting the proliferation of human hepatocarcinoma cell line HepG2

作　　者：赵艳阳 李伟[1] 郭伟[1] ZHAO Yan-yang;LI Wei;GUO Wei(Department of General Surgery Branch,Beijing Friendship Hospital,Capital Medical University,Beijing 100050,China)

机构地区：[1]首都医科大学附属北京友谊医院普外分中心,北京100050

出　　处：《临床和实验医学杂志》2021年第19期2017-2020,共4页Journal of Clinical and Experimental Medicine

基　　金：国家科技支撑计划课题(编号:2015BAI13B09)。

摘　　要：目的探讨环匹酮胺抑制人肝癌细胞株HepG2生长增殖和促进细胞凋亡作用。方法分别在24、48、72 h时间点以及3.125、6.25、12.5、25、50μmol/L浓度的环匹酮胺、5-氟尿嘧啶处理人肝癌细胞株HepG2,并设置对照组(空白处理)。用磺酰罗丹明B(SRB)比色法来观察对HepG2细胞数目的影响;倒置相差显微镜观察细胞形态学上的变化;膜联蛋白V(Annexin V)/碘化丙啶(PI)染色法来检测对细胞凋亡率的影响;蛋白质印迹法检测Bcl-2、cleavedcaspase-3、Bax凋亡相关蛋白表达水平。结果环匹酮胺处理24 h开始出现细胞皱缩、破碎以及数量减少,时间愈久细胞形态变化愈明显。环匹酮胺对人肝癌HepG2细胞增殖有抑制作用,这种抑制作用浓度愈高作用愈强,50μmol/L下作用最强,时间愈久作用愈强,在72 h下作用最强。环匹酮胺在3.125-50μmol/L均较同浓度下的5-氟尿嘧啶效果好。环匹酮胺可减少抗凋亡蛋白Bcl-2的表达,增加促凋亡蛋白cleavedcaspase-3、Bax表达。环匹酮胺可显著性地使肝癌细胞的细胞周期出现凋亡特异性的Sub-G1分期,引起肝癌的凋亡。结论环匹酮胺能够抑制人肝癌HepG2细胞增殖,引导细胞凋亡,改变肝癌细胞周期,其抗癌作用机制需进一步探究。Objective To explore the effect of cyclopiroxamine on inhibiting the growth and proliferation of human liver cancer cell line HepG2 and promoting cell apoptosis.Methods The human hepatoma cell line HepG2 was treated with cyclazone and 5-fluorouracil at 24,48,72 h time points and 3.125,6.25,12.5,25,and 50μmol/L,respectively,and a control group(blank treatment)was set up.The sulfonyl rhodamine B(SRB)method was used to observe the effect on the number of HepG2 cells.Inverted phase-contrast microscope to observe the changes in cell morphology;Annexin V/propidium iodide(PI)staining method to detect the influence on cell apoptosis rate;Western blotting method to detect the expression levels of Bcl-2,cleavedcaspase-3,Bax apoptosis-related proteins.Results Cyclopistonamine treatment began to appear cell shrinkage,fragmentation,and number reduction for 24 h.The longer the time,the more obvious the changes in cell morphology.Cyclopiroxamine has an inhibitory effect on the proliferation of human liver cancer HepG2 cells.The higher the concentration of this inhibitory effect,the stronger the effect,the strongest effect was at 50μmol/L,the longer the time the stronger the effect,and the strongest effect was at 72 h.Cyclopiroxamine was more effective than 5-FU at the same concentration at 3.125-50μmol/L.Cyclopiroxamine could reduce the expression of anti-apoptotic protein Bcl-2 and increase the expression of pro-apoptotic proteins leavedcaspase-3 and Bax.Cyclopiroxamine could significantly make the cell cycle of hepatocellular carcinoma cells appear apoptosis-specific Sub-G1 staging,and cause liver cancer apoptosis.Conclusion Cyclopiroxamine can inhibit the proliferation of human liver cancer HepG2 cells,guide cell apoptosis,and change the cycle of liver cancer cells.Its anti-cancer mechanism needs to be further explored.

关 键 词：肝癌细胞 环匹酮胺 增殖 凋亡 

分 类 号：R735.7[医药卫生—肿瘤]