miR-200a靶向调控间质表皮转化因子的表达参与肝癌细胞生物学行为  

miR-200a involvement in the biological behavior of hepatoma carcinoma cells by targeting the regulatory expression of mesenchymal-epithelial transition factor

作　　者：张亮[1] 陈威[1] 侯振国[1] 杨熙 刘明慧[2] Zhang Liang;Chen Wei;Hou Zhenguo;Yang Xi;Liu Minghui(Department of Radiology,Xingtai People's Hospital,Xingtai 054000,China;Physiology Teaching and Research Department of the Basic Department of Xingtai Medical College,Xingtai 054000,China)

机构地区：[1]邢台市人民医院放射科,邢台054000 [2]邢台医学高等专科学校基础部生理教研室,邢台054000

出　　处：《中华肝脏病杂志》2023年第11期1176-1181,共6页Chinese Journal of Hepatology

基　　金：河北省医学科学研究重点课题计划(20191715)。

摘　　要：目的研究miR-200a对间质表皮转化因子(MET)的调控作用及其对人肝癌细胞生物学行为的影响。方法采用荧光素酶报告系统确定miR-200a对MET的调控作用。将人肝癌HepG2细胞分为对照组、miR-200a组、MET过表达组及共转染组(miR-200a+MET)。经培养后,采用细胞计数(CCK-8)法检测细胞增殖能力,采用细胞划痕实验检测细胞迁移能力,采用Annexin V-FITC染色观察细胞凋亡情况,Transwell小室检测细胞侵袭能力,并采用蛋白质印迹法检测MET以及凋亡相关(Bcl-2、Caspase-3、Bax)蛋白的表达情况。两组间比较应用独立样本t检验,多组间比较采用单因素方差分析进行统计学分析。结果荧光素酶实验表明miR-200a可靶向MET。miR-200a组肝癌HepG2细胞增殖倍数、细胞侵袭个数(55.00±7.21、85.00±7.94、164.67±19.22、104.00±12.29)、划痕愈合率(28.33%±5.03%、61.67%±4.04%,74.67%±7.02%,49.33%±9.02%)以及MET、Bcl-2、Caspase-3蛋白表达水平均低于对照组、MET过表达组、共转染组,而MET过表达组各项指标高于其他3组,组间比较差异有统计学意义(P<0.05)。miR-200a组肝癌HepG2细胞凋亡率(19.25%±2.98%、6.80%±1.15%、3.42%±0.76%、9.90%±2.72%)、Bax蛋白表达水平高于对照组、MET过表达组、共转染组,而MIF过表达组各项指标水平低于其他3组,对照组、共转染组介于两者之间,组间比较差异有统计学意义(P<0.05)。结论miR-200a可抑制肝癌HepG2细胞增殖、迁移、侵袭,以及诱导细胞凋亡,其作用机制可能与miR-200a靶向下调肝癌HepG2细胞MET的表达有关。Objective To study the regulatory effect of miR-200a on mesenchymal-epithelial transition factor(MET)and its impact on the biological behavior of hepatoma carcinoma cells.Method A luciferase reporter assay was used to determine miR-200a's regulatory impact on MET.Human hepatoma HepG2 cells were divided into a control group,a miR-200a group,a MET overexpression group,and a co-transfection group(miR-200a+MET).After culture,cell proliferation ability,cell migration ability,apoptosis,cell invasion ability,and the expression of MET and apoptosis-related(Bcl-2,Caspase-3,Bax)proteins were detected and observed by cell counting kit-8(CCK-8),scratch assay,Annexin V-FITC staining,transwell chambers,and western blotting.The two groups were compared using the independent sample t-test.The multiple groups were statistically analyzed using one-way ANOVA.Results The luciferase experiment showed that miR-200a had target MET.The proliferation rate,number of invasions in cells(55.00±7.21,85.00±7.94,164.67±19.22,104.00±12.29),scratch healing rate(28.33%±5.03%,61.67%±4.04%,74.67%±7.02%,49.33%±9.02%),and expression levels of MET,Bcl-2,and Caspase-3 proteins were lower in the miR-200a group than those in the control group,MET overexpression group,and co-transfection group,while the MET overexpression group had higher indexes than the other three groups,with statistically significant differences between the groups(P<0.05).The apoptosis rate of HepG2 cells and the expression level of Bax protein were higher in the miR-200a group than those in the control group,MET overexpression group,and co-transfection group(19.25%±2.98%,6.80%±1.15%,3.42%±0.76%,9.90%±2.72%),while the levels of various indexes in the MIF overexpression group were lower than those in the other three groups.The control group and co-transfection group were between the two groups,and the difference between the groups was statistically significant(P<0.05).Conclusion HepG2 cell proliferation,migration,invasion,and cell apoptosis induction can be inhibited by miR-

关 键 词：肝癌 miR-200a 间质表皮转化因子 肝癌细胞 生物学行为 细胞凋亡 

分 类 号：R735.7[医药卫生—肿瘤]