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目的:探讨RNA干扰沉默NANOG表达后对肝癌细胞HepG2中细胞周期素D1(cyclin D1)表达及细胞增殖的影响。方法:将以NANOG基因为靶点的NANOG-siRNA瞬时转染肝癌细胞HepG2,real-time PCR和Western boltting检测NANOG、cyclin D1 mRNA和蛋白的表达,CCK-8和平板克隆形成实验检测细胞增殖能力,流式细胞术检测细胞周期情况。结果:与mock组比较,转染NANOG-siRNA后,肝癌细胞HepG2中NANOG、cyclin D1mRNA和蛋白水平均下调(P<0.05),细胞增殖能力下降(P<0.05),进入G0/G1期的细胞比例增多(P<0.05)。结论:沉默NANOG表达可引起肝癌细胞HepG2中cyclin D1的表达下降,导致细胞增殖能力下降。
AIM: To investigate the effect of RNA interference on the expression of cyclin D1 and cell proliferation in HepG2 hepatoma cells after NANOG expression. Methods: Nanog siRNA targeting NANOG gene was transiently transfected into HepG2 hepatoma cells. The expression of NANOG and cyclin D1 mRNA and protein was detected by real-time PCR and Western blotting. The proliferation of cells was detected by CCK-8 and plate clone formation assay , Flow cytometry to detect cell cycle. Results: Compared with the mock group, the mRNA and protein levels of NANOG, cyclin D1 and HepG2 in HepG2 cells were down-regulated (P <0.05), and the cell proliferation ability was decreased (P <0.05) The proportion increased (P <0.05). Conclusion: Silencing NANOG expression can decrease the expression of cyclin D1 in HepG2 hepatoma cells, leading to the decrease of cell proliferation.