cFLIP蛋白在大肠癌细胞中的表达及其对大肠癌细胞凋亡的影响

Expression of cFLIP in colorectal cancer and its modulation effect on the apoptosis of colon cancer cells

目的检测细胞型Fas相关死亡区域蛋白样β白细胞介素1转换酶抑制蛋白(cFLIP)在大肠癌细胞中的表达,探讨其在大肠癌细胞凋亡中的作用。方法应用Western蛋白印迹法检测3株人大肠癌细胞株中cFLIP蛋白的表达及大分子合成抑制剂放线菌素D(ActD,10μg/L)对SW620细胞cFLIP表达的影响,并运用四唑盐比色法(MTT法)检测cFLIP对SW620细胞凋亡的影响。结果SW1116及SW620细胞株cFLIP蛋白呈阳性表达,而LoVo细胞株呈阴性表达。ActD(10μg/L)可明显下调SW620细胞中cFLIP蛋白的表达水平,其作用1h以后,cFLIP表达水平开始下降(为0h的86%),作用24h后下降最明显(为0h的59%)。同时,抗Fas抗体(1mg/L)可最大程度地诱导28.8%的SW620细胞发生凋亡。结论cFLIP蛋白在大肠癌细胞中表达并可能在抑制大肠癌细胞凋亡中发挥重要作用。

Objective To investigate the expression of cFLIP in colorectal cancer cells at protein level and its modulation effect on the apoptosis of colon cancer cells.Methods Western blot was employed to detect the expression of cFLIP in 3 colorectal cancer cell lines and the effect of actinomycin D (Act D) (10 μg/L) on the cFLIP expression of SW620 cell line.Then the effect of cFLIP on apoptosis in SW620 cell line pretreated with Act D was examined by MTT assay.Results cFLIP was positively expressed in SW1116 and SW620 cell lines and negatively in LoVo cell line.Act D (10 μg/L) treatment can obviously decrease the expression levels of cFLIP in SW620 cell line.cFLIP levels were found to decrease within 1 h after addition of Act D (86% of 0 h) and were significantly reduced after 24 h (59% of 0 h).At the same time,the anti Fas (1 mg/L) could induce 28.8% apoptosis of SW620 cell.Conclusion cFLIP is expressed in some colorectal cancer cell lines and may play an important role in the inhibition of apoptosis induced by CD95.