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二甲双胍与顺铂联合诱导人结肠癌HCT-8细胞凋亡的研究 预览

Induction of apoptosis in human colon cancer HCT-8 cells by metformin in combination with cisplatin
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摘要 目的研究二甲双胍联合顺铂对HCT-8人类结肠癌细胞凋亡的影响及其可能机制。方法将HCT-8人类结肠癌细胞分成二甲双胍组(MET)、顺铂组(DDP)、二甲双胍联合顺铂组(MET+DDP)、空白对照组(CK)4组,MTT实验观察4组细胞于24 h、48 h、72 h时的增殖能力;流式细胞术观察各组细胞48 h的凋亡比例;Western blotting观察各组细胞48 h时凋亡相关蛋白Bcl-2、Bax、caspase-3(激活型)以及AKT/GSK-3通路蛋白的表达水平。结果该实验中MET+DDP组的细胞增殖能力弱于其它三组(P<0.05);细胞凋亡比例高于其它三组(P<0.05);Bax、caspase-3(激活型)表达量高于其它组群,Bcl-2、P-AKT、P-GSK-3表达量低于其它组群,AKT、GSK-3在4组中表达量相对恒定(P >0.05)。结论二甲双胍、顺铂可能通过下调AKT/GSK-3信号通路,改变Bcl-2家族蛋白的表达活性从而促进HCT-8人类结肠癌细胞的凋亡,并促进p-caspase-3剪切转化为caspase-3(激活型);二甲双胍与顺铂可协同发挥促HCT-8人类结肠癌细胞凋亡的作用。 Objective To investigate the effects and mechanisms of action of metformin combined with cisplatin on the apoptosis in human colon cancer HCT-8 cells.Methods Human colon cancer HCT-8 cells were divided into four groups:metformin group(MET),cisplatin group(DDP),metformin combined with cisplatin group(MET+DDP),and blank control group(CK).The proliferative ability of HCT-8 cells was determined by an MTT experiment at24,48 and 72 h.The apoptosis rate was determined by flow cytometry at 48 h.Western blotting was used to reveal the expression of apoptosis-related proteins such as bcl-2,Bax and caspase-3(activated),as well as proteins of the AKT/GSK-3βpathway at 48 h.Results The cell proliferative ability in the MET+DDP group was weaker than that of the other three groups(P<0.05).Moreover,its apoptosis rate was higher than those of the other three groups(P<0.05).The expressions of Bax and caspase-3(activated)was higher than that of the other groups,the expression of Bcl-2,p-AKT and p-GSK-3βwas lower than that of the other groups,and the expression of AKT and Gsk-3 was relatively consistent among the four groups.Conclusions Metformin and cisplatin in combination can reduce the activity of the AKT/GSK-3βsignaling pathway,and change the expression of Bcl-2 family proteins,to promote the apoptosis of human colon cancer HCT-8 cells.They also promote the p-caspase-3 conversion to caspase-3 p-caspase-3 conversion to caspase-3(activated),metformin and cisplatin can thus play a role in promoting the apoptosis of HCT-8 human colon cancer cells.
作者 于敬坤 李明志 随振阳 张琪 田炜 胡琨 杨光华 张国志 王长友 YU Jingkun;LI Mingzhi;SUI Zhenyang;ZHANG Qi;TIAN Wei;HU Kun;YANG Guanghua;ZHANG Guozhi;WANG Changyou(Department of General Surgery,Affiliated Hospital of North China University of Science and Technology,Tangshan,Hebei,063000,China;Medical research center,North China University of Science and Technology,Tangshan,Hebei,063000,China)
出处 《中国比较医学杂志》 CAS 北大核心 2019年第2期43-50,共8页 Chinese Journal of Comparative Medicine
基金 中国煤炭工业协会2017年度科学技术研究指导性计划项目(MTKJ2017-331)。
关键词 HCT-8人类结肠癌细胞 二甲双胍 凋亡 AKT/GSK-3β信号通路 human colon cancer HCT-8 cell metformin apoptosis AKT/GSK-3βsignaling pathways
作者简介 于敬坤(1991-),男,山东聊城,硕士研究生,研究方向:结肠癌;E-mail:1843293446@qq.com;通信作者:王长友(1971-),男,河北唐山,硕士,主任医师,研究方向:胃肠外科学。E-mail:fhbj-2004@163.com.
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