Sp3表达下调后肝癌细胞株HepG2基因表达谱的变化

Alteration of gene expression profile in hepatocellular carcinoma HepG2 cells with Sp3 downregulation

目的:分析转录因子Sp3表达下调后人肝癌细胞株HepG2基因表达谱的变化.方法:使用siRNA干扰技术下调人肝癌细胞株HepG2中Sp3表达,人全基因组表达谱芯片筛查Sp3沉默后的差异表达基因,qRT-PCR验证部分差异表达基因,流式细胞术检测细胞周期.结果:Sp3表达下调后,筛查出差异表达基因1789个,其中上调基因1007个,下调基因782个,涉及细胞增殖、分化、凋亡、黏附、代谢等多方面功能,qRT-PCR结果表明周期相关基因CCND1、CCNE2、转化生长因子B1(transforming growth factor B1,TGFB1)、CDKN2A的表达变化与芯片结果一致,流式细胞术结果显示Sp3表达下调后细胞主要分布在G1期.结论:Sp3表达下调后人肝癌细胞株HepG2的基因表达谱发生明显变化,并出现G0/G1期阻滞,Sp3可能通过细胞周期因子参与肝癌的发生发展过程.

AIM： To analyze the alteration of gene expression profile in hepatocellular carcinoma cell line HepG2 with Sp3 downregulation. METHODS： RNA interference was performed to downregulate Sp3 mRNA in HepG2 cells and NimbleGen Human Gene Expression Microarray was used for gene expression profile analysis. Real-time quantitative PCR was employed to confirm the expression of several differentially expressed genes. The cell cycle was analyzed by flow cytometry. RESULTS： A total of 1789 genes was found to be differentially expressed in HepG2 cells with Sp3 downregulation, including 1007 up-regulated and 782 down-regulated ones. These genes were involved in many cellular biological process such as proliferation, differentiation, programmed death, adhesion, and metabolic process. Real-time quantitative PCR confirmed the alteration of several cell cycle related genes （CCND1, CCNE2, TGFB1, and CDKN2A）. Flow cytometry analysis showed that the percentage of cells in G1 phase increased significantly after Sp3 downregulation.CONCLUSION： Gene expression profile alters in HepG2 with Sp3 downregulation. Sp3 knockdown induces cell cycle arrest in G0/G1 phase in HepG2 cells. Sp3 may play an essential role in the pathogenesis of hepatocellular carcinoma as a transcription factor via regulating cell cycle progression.