摘要
目的探讨miR-218是否通过调控含有4个mbt区域的scm相关基因(SFMBT1)影响肝癌细胞增殖能力。方法 RT-PCR检测人肝癌细胞Hu H-7、Hep G2、SNU-449和人正常肝细胞HH中miR-218的水平。靶基因预测软件预测miR-218的靶基因并用荧光素酶报告基因进行鉴定,RT-PCR和Western blot检测miR-218对靶基因的调控作用。肝癌细胞转染miR-218模拟物(miR-218 mimics组)、SFMBT1小干扰RNA(SFMBT1 siRNA组),同时设置对照组(只加入转染试剂)。噻唑蓝(MTT)检测各组细胞存活率。结果肝癌细胞Hu H-7、Hep G2、SNU-449中miR-218表达水平均明显低于正常肝细胞HH(P<0.01)。Hep G2细胞中miR-218表达水平最低,后续选用Hep G2细胞深入研究。miR-218的靶基因是SFMBT1,并且miR-218负调控SFMBT1靶基因。miR-218 mimics组和SFMBT1 siRNA组细胞存活率均明显低于对照组(P<0.01)。miR-218 mimics和SFMBT1 siRNA共转染后的细胞存活率比miR-218mimics组和SFMBT1 siRNA组均明显降低(P<0.01)。结论 miR-218能够负调控靶基因SFMBT1抑制人肝癌细胞的增殖。
Objective To investigate whether miR-218 can regulate SFMBT1 gene to affect the proliferation of hepatocellular carcinoma cells.Methods RT-PCR was used to detect the level of miR-218 in human hepatocellular carcinoma cell line HuH-7,HepG2,SNU-449 and human normal liver cell line HH.Target gene prediction software were used to predict the target gene of miR-218 and to identify by the luciferase reporter gene,to detect the effect of miR-218 on target gene by RT-PCR and Western blot.Hepatocellular carcinoma cells were transfected with miR-218 mimics,SFMBT1 siRNA group,and the control group was set up,while the control group was only transfected with transfection reagent,MTT was used to detect the survival rate of each group.Results The expression of miR-218 in human hepatocellular carcinoma cells HuH-7,HepG2,SNU-449 was significantly lower than that of the control group （P＆lt;0.01）.The miR-218 target gene was SFMBT1 and miR-218 negatively regulated the SFMBT1 target gene.The expression level of miR-218 in HepG2 cells was the lowest,followed study with HepG2 cells.The survival rates of miR-218 group and SFMBT1 siRNA group were significantly lower than those of control group （mimics）,respectively （P＆lt;0.01）.The cell survival rate of miR-218 mimics and SFMBT1 siRNA was significantly lower than that of miR-218 mimics group and SFMBT1 siRNA group （P＆lt;0.01）.Conclusion miR-218 can inhibit the proliferation of human hepatocellular carcinoma cell line by negative regulation of target gene SFMBT1.
出处
《临床和实验医学杂志》
2017年第13期1272-1275,共4页
Journal of Clinical and Experimental Medicine
