摘要
目的探讨miR-548a-5p调控肝癌细胞HepG2凋亡的潜在机制。方法通过miR-548a-5p mimics和miR-548a-5p inhibitor分别过表达和敲低miR-548a-5p,检测HepG2的凋亡水平。通过miRDB在线分析和荧光素酶报告系统检测miR-548a-5p的潜在底物。通过siRNA和PCDNA3.1载体过表达底物,检测HepG2的凋亡水平。结果过表达miR-548a-5p后,HepG2的凋亡水平下降,miR-548a-5pd的潜在靶向HMBOX1的表达量显著降低;相反,在敲低miR-548a-5p后,HepG2的凋亡水平上升,HMBOX1的表达量显著上升(P<0.05)。荧光素酶报告系统显示miR-548a-5p靶向HMBOX1的3端非编码区,在敲低HMBOX1后,HepG2的凋亡水平明显下降,过表达后则相反(P<0.05)。结论miR-548a-5p通过靶向HMBOX1的mRNA的3端非编码区抑制HMBOX1的翻译,从而抑制肝癌细胞HepG2的凋亡。
Objective To investigate the potential mechanism of mir-548a-5p regulating the apoptosis of hepatocellular carcinoma(HCC)cell line HepG2.Methods MiR-548a-5p mimics and miR-548a-5p inhibitor were used to overexpress and knockdown miR-548a-5p and the apoptosis levels of HepG2 were detected.Potential substrates of miR-548a-5p were detected by miRDB online analysis and luciferase reporting system.The siRNA and PCDNA3.1 vector were used to overexpress substrates and the apoptosis level of HepG2 was detected.Results After overexpressing miR-548a-5p,the apoptosis level of HepG2 decreased,and the expression level of HMBOX1,a potential target of miR-548a-5p,decreased significantly.On the contrary,after knocking down miR-548a-5p,the apoptosis level of HepG2 increased,and the expression of HMBOX1 increased significantly(P<0.05).Luciferase reporter system showed that miR-548a-5p targeted the 3-terminal non-coding region of HMBOX1.After knocking down HMBOX1,the apoptosis level of HepG2 decreased significantly,and vice versa(P<0.05).Conclusion MiR-548a-5p inhibits the translation of HMBOX1 by targeting the 3-terminal non-coding region of HMBOX1 mRNA,thereby inhibiting the apoptosis of HepG2 of HCC cells.
作者
袁溢苒
陈龙
张臻
YUAN Yi-ran;CHEN Long;ZHANG Zhen(Nanchong Central Hospital Special Medical Department,Nanchong,Sichuan 637000,China;Department of Digestive Medicine,Nanchong Central Hospital;Nanchong Health Committee Nanchong,Sichuan 637000,China)
出处
《肝脏》
2021年第8期871-873,共3页
Chinese Hepatology
