GFER抑制四氯化碳对HepG2细胞的损伤

GFER Restrains Injury Induced by CCl_4 in HepG2 Cells

目的探讨HepG2细胞内生长因子ERV1样基因(growth factor Erv1-gene,GFER)表达降低后对四氯化碳(CCl4)诱导的细胞损伤的影响,以进一步明确GFER对于肝细胞的保护作用。方法首先将GFER siRNA转染入HepG2细胞,72 h后收集细胞并通过Western Blot检测GFER的表达以明确沉默效率。再次将GFER siRNA转染入HepG2细胞72 h后,用CCl4处理细胞6 h和24 h,检测细胞内ATP的含量,caspase-3的活性,并应用MTS方法测定细胞的增殖能力以及TUNEL方法检测细胞凋亡。结果 Western blot结果显示转染GFER siRNA后细胞内GFER的表达降低。CCl4处理细胞6 h后,GFER表达降低使细胞的增殖能力下降,细胞内ATP含量增加,细胞凋亡更为明显。CCl4处理24 h后,GFER表达降低使细胞的增殖能力进一步下降,Caspase 3活性进一步升高,凋亡细胞数目显著增多,而ATP的含量明显下降。结论 GFER表达降低促进CCl4对HepG2细胞的损伤。

Objective To explore the effects of downregulation of GFER gene expression on cell injury induced by CCl4 in HepG2 cells so as to elucidate the protective effect of GFER on hepatocytes. Methods GFER siRNA was transfected into HepG2 cells for 72 h,and western blot was used to detect the protein level of GFER in order to assess transfection efficiency. After 72 h of GFER siRNA interference,the HepG2 cells were treated with CCl4 for 6 h and 24 h. ATP levels and caspase-3 activity were measured. MTS assay was used to measure cell proliferation. TUNEL was used to detect and quantify apoptotic cell death. Results Western blot analysis showed that the protein level of GFER was decreased after GFER siRNA transfectioin. Cell proliferation of HepG2 cells was inhibited by GFER siRNA interference,while the ATP levels and the number of apoptotic cells were both increased after 6 h of CCl4 treatment in the siRNA intervention group as compared to the controls. When cells were treated with CCl4 for 24 h,proliferation of HepG2 cells was further inhibited after siRNA intervention,and the caspase-3 activity and the number of apoptotic cells were further increased compared to the results obtained at 6 h after CCl4 treatment. However,the ATP levels in HepG2 cells were markedly decreased. Conclutsion Downregulation of GFER gene expression aggravates HepG2 cell injury induced by CCl4.