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目的探讨癌基因△Np63对膀胱癌细胞增殖和凋亡的影响,并初步探讨其可能的分子机制。方法通过将△Np63特异性短发夹RNA(△Np63-shRNA)和非特异性短发夹RNA(Control-shRNA)转染膀胱移行细胞癌(transitional cell carcinom a of bladder,TCCB)5637细胞,用半定量逆转录-聚合酶链反应(RT-PCR)和Westernblot分别检测△Np63、p27kip1、p57kip2mRNA和蛋白的表达水平,WST-1法检测细胞增殖活性,利用流式细胞术(FCM)检测细胞周期分布,TUNEL法检测细胞凋亡情况。结果特异性的△Np63-shRNA能够有效地沉默△Np63并上调p27kip1和p57kip2的基因和蛋白水平,转染△Np63-shRNA的5637细胞的增殖能力明显受到抑制(P<0.05),且明显促进了细胞的凋亡(P<0.05)。结论靶向△Np63基因的shRNA片段可以有效的抑制人膀胱癌5637细胞的增殖并促进其凋亡,其可能的机制是通过上调p27kip1和p57kip2的表达实现的。
Objective To investigate the effects of oncogene △ Np63 on proliferation and apoptosis of bladder cancer cells and to explore its possible molecular mechanism. METHODS: Transfected cells were transfected with △ Np63 specific short hairpin RNA (△ Np63-shRNA) and non-specific short hairpin RNA (shRNA-shRNA) into transitional cell carcinom of a bladder (TCCB) The expression of △ Np63, p27kip1 and p57kip2 mRNA and protein were detected by quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The cell proliferation activity was detected by WST-1 method. The cell cycle distribution was detected by flow cytometry (FCM) , TUNEL assay apoptosis. Results The specific Np63-shRNA could effectively silence △ Np63 and up-regulate the gene and protein levels of p27kip1 and p57kip2. The proliferation of 5637 cells transfected with △ Np63-shRNA was significantly inhibited (P <0.05) and significantly promoted Cell apoptosis (P <0.05). Conclusion The shRNA targeting △ Np63 gene can effectively inhibit the proliferation and promote the apoptosis of human bladder cancer cell line 5637. Its possible mechanism is through up-regulating the expression of p27kip1 and p57kip2.