MicroRNA-33b通过靶向调控SALL4的表达抑制肝细胞癌细胞增殖

Micro RNA-33b inhibits cell proliferation in hepatocellular carcinoma via targeting SALL4

目的:探索微小RNA-33b(micro RNA-33b,mi R-33b)在肝细胞癌(hepatocellular carcinoma,HCC)中的表达及调控HCC细胞增殖的相关分子机制。方法:收集配对的HCC及癌旁组织32对,分别采用实时荧光定量PCR和Western印迹法检测人类婆罗双树样基因4(Sal-like 4,SALL4)RNA和蛋白表达,MTT实验检测HCC细胞增殖,萤光素酶报告基因检测验证mi R-33b与SALL4基因的靶点关系。结果:Mi R-33b在HCC组织中的表达显著低于癌旁组织(P<0.001)。过表达mi R-33b能显著降低HCC LH86细胞的增殖。SALL4是mi R-33b的靶基因,其蛋白表达被mi R-33b负调控。过表达SALL4能逆转mi R-33b对LH86细胞增殖的抑制作用。SALL4在HCC组织中的表达显著高于癌旁组织(P<0.001)。结论:Mi R-33b对HCC细胞增殖的抑制作用是通过负调控SALL4的表达而实现。

Objective: To investigate the expression of miR-33 b in hepatocellular carcinoma(HCC) and to explore regulatory mechanism of mi R-33 b for cell proliferation of HCC.Methods: HCC tissues and adjacent non-tumor tissues were collected for this study(n=32 for each). Real-time PCR and Western blot were conducted to examine the mR NA and protein expression, respectively. MTT assay was used to detect the cell proliferation. Luciferase reporter gene assay was performed to verify the target relationship between mi R-33 b and Sal-like 4(SALL4).Results: MiR-33 b was significantly downregulated in HCC tissues compared with adjacent non-tumor tissues. Overexpression of miR-33 b decreased the proliferation of HCC LH86 cells. SALL4 was identified as a target gene of miR-33 b, and its protein expression was negatively regulated by miR-33 b. Overexpression of SALL4 reversed the suppressive effect of miR-33 b on LH86 cell proliferation. SALL4 was significantly upregulated in HCC tissues compared with adjacent nontumor tissues. Conclusion: The miR-33 b suppresses HCC cell proliferation through down-regulation of SALL4.