基因fas的表达对细胞色素C诱导HL-60细胞凋亡的影响

Effect of fas mRNA Expression on Cytochrome C-induced Apoptosis of HL-60 Cells In Vitor

本研究观察fas基因表达的变化对细胞色素C诱导急性早幼粒细胞白血病细胞株HL-60细胞凋亡的影响,并探讨细胞色素C诱导HL-60细胞凋亡的机制。将体外培养的HL-60细胞分成6组,细胞色素C终浓度分别为0(对照组)、9.375、18.75、37.5、75、150mg/L,用流式细胞仪检测细胞色素C对HL-60细胞的凋亡作用,用RT-PCR和Western blot检测以凋亡为主的HL-60细胞中fas的表达水平。结果表明:细胞色素C终浓度分别为0、9.375、18.75及37.5mg/L时HL-60细胞是以凋亡效应为主,fas基因的mRNA和蛋白的表达随着细胞色素C浓度的增高而增高,当细胞色素C浓度为75mg/L、150mg/L时,HL-60细胞是以坏死效应为主。结论:细胞色素C能够上调fas mRNA及其蛋白的表达,从而可以诱导HL-60细胞的凋亡。

The aim of this study was to investigate the effect of cytochrome C on apoptosis of HL-60 cells and to explore the mechanism of HL-60 cell apoptosis induced by cytochtome C. The HL-60 cells were treated with different concentlations of cytochrome C for 24 hours. The concentrations of cytochrome C were as follows： 0 （control group）, 9. 375 mg/L, 18.75 mg/L, 37.5 mg/L, 75 mg/L and 150 mg/L, The apoptosis was analyzed by flow cytometry （FCM） after HL-60 cells were treated with different concentrations of cytochrome C for 24 hours, The fas mRNA expression changes of relevant apoptotic genes were examined by reverse transcription-polymerase chain reaction （ RT- PCR） after HL-60 cells were treated with different concentrations of cytochrome C for 24 hours and were analyzed halfquantitatively. The expressions of fas involved in HL-60 treated with different concentrations of cytochtome C for 24 hours were detected by Western blot. The results indicated that the flow cytometry （FCM） （PI straining） showed obvious hypo-diploid peak before Gt phase in the treated group, and its apoptotic ratio increased in a dose-dependent manner. The fas mRNA expression levels of the relevant apoptotic genes could be detected by RT-PCR after HL-60 cells treated with different concentrations of cytochrome C for 24 hours, and the expression of fas mRNA in HL-60 cells was increased by cytochrome C in dose-dependent manner within range of 0 - 37.5 mg/L. It is concluded that the cytochrome C up-regulates expressions of fas mRNA and their protein so as to induce apoptosis of the HL-60 cells.