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目的:比较不同大肠癌细胞Fas受体及其下游通路抑制因子FLIP的表达差异,探讨Fas、FLIP对大肠癌细胞凋亡的影响. 方法:体外培养大肠癌细胞,应用直接免疫荧光流式细胞术检测细胞表面Fas表达率,半定量RT-PCR法测定FLIP mRNA的水平,并采用Annexin V法评价细胞对Fas介导的凋亡的敏感性. 结果:大肠癌细胞表面Fas表达率不同,其中HT-29细胞的表达率为42.46±4.32%,明显高于其他3株细胞.SW620 和HT-29细胞FLIP mRNA含量较高,Colo205居中,Lovo 则呈阴性;且相对于任何一株FLIP表达阳性的细胞,FLIPL的表达水平均高于FLIPs.给予凋亡诱导型抗Fas抗体(CH-11)刺激后,4株大肠癌细胞凋亡敏感性均较低. 结论:不同的大肠癌细胞株可能通过不同途径逃避Fas介导的凋亡,其中包括下调Fas的表达和上调FLIP的含量.
OBJECTIVE: To compare the expression of Fas receptor and its downstream pathway inhibitor FLIP in different colorectal cancer cell lines, and to explore the effect of Fas and FLIP on the apoptosis of colorectal cancer cells.Methods: Colorectal cancer cells were cultured in vitro and cultured with direct immunofluorescence flow cytometry The rate of Fas expression on the cell surface was measured, the level of FLIP mRNA was detected by semi-quantitative RT-PCR, and the sensitivity of Fas-mediated apoptosis was evaluated by Annexin V.Results: The expression of Fas on the surface of colorectal cancer cells was different, -29 cells was 42.46 ± 4.32%, which was significantly higher than that of the other 3 cells.The FLIP mRNA content was higher in SW620 and HT-29 cells, Colo205 was middle and Lovo was negative, The expression level of FLIPL and FLIPL were higher than that of FLIPs.Apoptosis of 4 colorectal cancer cells was lower than that of FLIPs.Conclusion: Different colorectal cancer cell lines may pass Different pathways evade Fas-mediated apoptosis, including down-regulation of Fas expression and up-regulation of FLIP levels.