Notch1表达对肝癌细胞HepG2生物学行为的影响

Effect of Notch1 expression on biological behaviour of human hepatocarcinoma cell HepG2

目的研究Notch1基因对Hes1、Hes2表达水平以及肝癌细胞HepG2生物学行为的影响。方法将HepG2细胞分为Notch1受体胞内段质粒(A)组、Notch1干扰RNA(B)组、空载体(C)组和未处理HepG2细胞(D)组。采用MTT法检测各组细胞增殖能力,流式细胞术测定细胞周期,RT-PCR和Western blot法分别检测Notch1、Hes1、Hes2mRNA和蛋白的表达。结果与C组和D组相比,A组细胞增殖能力减弱,G1期细胞以及Notch1、Hes1、Hes2mRNA和蛋白表达均明显增加(P<0.05),而B组细胞增殖能力增强,G1期细胞以及Notch1、Hes1、Hes2mRNA和蛋白表达均明显减少(P<0.05)。结论上调Notch1的表达能促进Hes1与Hes2表达水平并抑制肝癌细胞HepG2增殖。

Objective To investigate the effects of Notchl gene on the expressions of Hesl and Hes2 and the biological behaviour of human hepatocarcinoma cell HepG2. Methods HepG2 cells were divided into four groups of A （transfected with exogenous intracellular domain of Notchl ）, B （transfected with Notch1 siRNA）, C（transfected with empty vector） and D（without transfection）. The cell proliferation and cell cycle were measured by MTT assay and flow cytometry. The mRNA and protein expressions of Notchl, Hesl and Hes2 were detected by RT-PCR and Western blot, respectively. Resdts Compared with groups of C and D, the cell proliferation was decreased, while the cells in Cn stage and the mRNA and protein expressions of Notchl, Hesl and Hes2 were increased in group A （P〈0. 05）, which were all reversed in group B （P〈0. 05）. Conclusion Upregulated expression of Notch1 gene can promote the levels of Hesl and Hes2 and inhibit the proliferation of hepatocarcinoma cell HepG2.