miR-1246在肝癌血清中的表达及对肝癌 HepG2细胞生物学功能的影响

Expression of microRNA-1246 in the serum of hepatocellular carcinoma and its effect on biological function of HepG2 cells

目的探讨miR-1246在肝癌患者血清中的表达及其对HepG2细胞生物学功能的影响。方法应用荧光定量PCR检测miR-1246在正常对照者和肝癌患者血清中的表达;将miR-1246 mimics和NC mimics分别转染至HepG2细胞,采用Transwell小室法、划痕实验和细胞克隆形成实验检测miR-1246对HepG2细胞生物学功能的影响,荧光定量PCR检测NFE2L3、Vimentin、GAS1基因的表达改变。结果 miR-1246在正常对照者和肝癌患者血清中的表达量分别为1.079(0.740,1.534)、0.132(0.068,0.324),差异有统计学意义(P<0.001);miR-1246 mimics转染HepG2细胞后,HepG2细胞穿过基质胶的细胞数为128.000±9.900,较NC mimics组(251.500±24.749)明显减少(P=0.023),划痕48 h后HepG2细胞相对迁移率为(24.833±3.351)%,较NC mimics组(31.859±7.194)%明显减慢(P=0.046),Vimentin、GAS1、NFE2L3基因相对表达量分别为0.318±0.213、0.479±0.157、0.755±0.044,较NC mimics组(1.000±0.000)表达均明显下调(P=0.046、0.043、0.030)。结论肝癌患者血清中miR-1246表达下调;miR-1246可能通过下调Vimentin、GAS1、NFE2L3基因抑制HepG2细胞的侵袭、迁移能力。

Objective To investigate the expression of microRNA-1246(miR-1246)in hepatocellular carcinoma(HCC)serum and its effect on the biological function of HepG2 cells.Methods Serum miR-1246 was determined by qRT-PCR in HCC patients and healthy controls.MiR-1246 mimics and NC mimics were transfected into HepG2 cells separately.The effect of miR-1246 on the biological function of HepG2 cells was examined by transwell chamber assay,scratch test and cell clone formation test.The expression of NFE2L3,Vimentin and GAS1 were detected by qHT-PCH.Results The expression levels of miR-1246 in the serum of healthy controls and HCC patients were 1.079(0.740,1.534)and 0.132(0.068,0.324),respectively,and the difference was statistically significant(P<0.001).After transfection of miR-1246 mimics into HepG2 cells,the number of HepG2 cells passing through matrigel was(128.000±9.900),which was significantly lower than that of the NC mimic group(251.500±24.749,P=0.023).Forty-eight hours after scratch,the relative mobility of HepG2 cells was(24.833±3.351)%,which was significantly lower than that in the INC mimic group(31.859±7.194)%(P=0.046).The relative expression levels of Vimentin,GAS1 and NFE2L3 were(0.318±0.213),(0.479±0.157)and(0.755±0.044),respectively,which were significantly lower than those in the NC mimic group(P=0.046,P=0.043 and P=0.030,respectively).Conclusion The expression levels of miR-1246 were decreased in HCC serum.MiR-1246 could inhibit the invasion and migration of HepG2 cells by down-regulating the expression of Vimentin,GAS1 and NFE2L3.