耐受TRAIL肾细胞癌株的建立及其与cFLIP蛋白的关系

The establishment of TRAIL-resistant renal cancer cell line and the relationship between it and cFLIP protein

目的建立耐受肿瘤坏死因子相关凋亡诱导配体(tumor necrosis factor-related apop-tosis-inducing ligand,TRAIL)的肾细胞癌株,观察Fas相关死亡域样白介素21β转换酶抑制蛋白(cFLICEinhibitory protein,cFLIP)与TRAIL耐药的关系。方法应用TRAIL间歇诱导方法建立耐受TRAIL的肾细胞癌株786-0TR,检测耐药株的半数抑制率(IC50)和耐药指数(RI);RT-PCR方法检测敏感株和耐药株cFLIP mRNA的表达,Western blot检测cFLIP蛋白的表达,并加以比较。结果 786-0TR细胞对TRAIL的IC50为739.3472μg/L,RI为9.6427;耐药株中cFLIPmRNA和蛋白的表达明显高于敏感株,二者之间差异有统计学意义(P<0.05)。结论成功建立了耐受TRAIL的肾细胞癌株;cFLIP蛋白是肾细胞癌对TRAIL耐药的重要因素。

Objective To establish the TRAIL-resistant renal cancer cell line and investigate the relationship between resistance and cFLIP protein.Methods A resistant cell line 786-0TR was established by stepwise increasing dose of TRAIL and intermittent administration.The resistant cell line IC50 and resistant index were measured.The cFLIP mRNA and protein of the resistant cell line and sensitive cell line were measured by RT-PCR and western blot.Then we compared the result.Results IC50 of resistant cell line was 739.347 2 μg/L,and its resistant index was 9.642 7.The expression of cFLIP mRNA and protein in resistant cell line higher than sensitive cell line （P0.05）.Conclusions We established the TRAIL resistant renal cancer cell line successively.cFLIP protein play an important role in renal cell carcinoma resisiting TRAIL.