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目的探讨肿瘤坏死因子相关凋亡诱导配体(TRAIL)对膀胱癌细胞的抑制作用。方法构建TRAIL融合增强绿色荧光蛋白(EGFP)真核表达载体,脂质体法转染膀胱癌细胞株EJ细胞,荧光显微镜下计算转染率。分别采用逆转录-聚合酶链反应(RT-PCR)、Western blot检测 TRAIL的表达。采用流式细胞仪检测细胞凋亡,噻唑蓝(MTT)法、细胞倍增时间和集落形成率观察TRAIL的抑制作用。结果 48 h观察重组载体转染率为38．4％,空载体转染率为35．3％(P> 0．05),RT-PCR和Western blot证实转染后EJ细胞中TRAIL的表达明显增加;凋亡率明显高于转染空白载体和未转染组(P<0．01);EJ细胞增殖受到明显抑制(P<0．01)。结论 TRAIL具有诱导膀胱癌细胞凋亡、抑制增殖的作用,有望成为治疗膀胱癌的新方法。
Objective To investigate the inhibitory effect of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) on bladder cancer cells. Methods The eukaryotic expression vector of TRAIL fusion enhanced green fluorescent protein (EGFP) was constructed and transfected into bladder cancer cell line EJ cells by lipofectamine. The transfection efficiency was calculated under fluorescence microscope. TRAIL expression was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot respectively. The inhibitory effect of TRAIL was observed by flow cytometry (FCM) and MTT assay, cell doubling time and colony formation rate. Results After 48 h, the transfection efficiency of recombinant vector was 38.4%, and the empty vector transfection rate was 35.3% (P> 0.05). The expression of TRAIL in transfected EJ cells was confirmed by RT-PCR and Western blot (P <0.01). The proliferation of EJ cells was significantly inhibited (P <0.01). Conclusion TRAIL can induce the apoptosis of bladder cancer cells and inhibit the proliferation of human bladder cancer cells. It is expected to become a new method for the treatment of bladder cancer.