论文部分内容阅读
目的:探讨DNMT1、DNMT3b基因在膀胱癌发生发展中的作用。方法:①将DNMT1、DNMT3bsiRNA重组质粒分别单独和共同转入人膀胱癌细胞系BIU-87;②半定量RT-PCR和Western Blot分别检测DNMT1和DNMT3b基因mRNA和蛋白水平的变化;③用MTT法观察各组细胞生长曲线,流式细胞术观察细胞生长周期及凋亡率的变化。结果:单独和共同转染DNMT1(或DNMT3b)mRNA、蛋白表达量明显下降,能影响BIU-87的生长曲线,使细胞增殖减慢,并使细胞周期阻滞于G_1期,明显提高细胞调亡率;转染空质粒和单独转染DNMT3b siRNA重组质粒对BIU-87细胞的生长和增殖无明显影响。结论:联合转染DNMT1、DNMT3b siRNA重组质粒,可同时下调DNMT1和DNMT3b在BIU-87细胞中的表达,并能在体外抑制BIU-87细胞生长,促进细胞凋亡发生,其效果优于单独转染的重组质粒。单独转染DNMT3b siRNA重组质粒对BIU-87细胞的生长和增殖无明显影响。
Objective: To investigate the role of DNMT1 and DNMT3b genes in the development of bladder cancer. Methods: ①The DNMT1 and DNMT3bsiRNA recombinant plasmids were separately and co-transfected into human bladder cancer cell line BIU-87; ② The mRNA and protein levels of DNMT1 and DNMT3b were detected by semi-quantitative RT-PCR and Western Blot; The growth curve of each group was observed. The cell growth cycle and apoptosis rate were observed by flow cytometry. Results: The DNMT1 (or DNMT3b) mRNA expression alone or co-transfected with DNMT1 (or DNMT3b) significantly decreased the expression of BIU-87 and inhibited the proliferation of BIU-87 cells and arrested the cell cycle in G_1 phase The transfected empty plasmid and DNMT3b siRNA plasmid alone had no significant effect on the growth and proliferation of BIU-87 cells. CONCLUSION: The co-transfection of DNMT1 and DNMT3b siRNA recombinant plasmids can both down-regulate the expression of DNMT1 and DNMT3b in BIU-87 cells and inhibit the growth and induce the apoptosis of BIU-87 cells in vitro Dyed recombinant plasmid. DNMT3b siRNA transfection alone had no significant effect on the growth and proliferation of BIU-87 cells.