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目的:探讨胃癌细胞中miR-193b对根据靶基因预测选出的纤溶酶原活化因子(uPA)的调控作用及机制。方法:胃癌BGC823细胞分别瞬时转染miR-193b正义序列(正义序列转染组),反义序列(反义序列转染组)及无意义序列(阴性转染组),以无转染的BGC823细胞为空白对照。用real-time PCR检测各组转染后miR-193b的表达水平,RT-PCR和Western blot法检测各组细胞uPA mRNA和蛋白的表达水平。结果:与空白对照组比较,阴性转染组miR-193b表达水平未见明显改变(P>0.05),正义序列转染组的miR-193b的表达水平明显上调,反义序列转染组miR-193b的表达水平明显下调(均P<0.05);各转染组uPA mRNA表达水平均无明显变化;阴性转染组uPA蛋白表达水平无变化,正义序列转染组uPA蛋白表达水平降低,反义序列转染组uPA蛋白表达水平增加。结论:uPA可能是miR-193b的靶基因,miR-193b可能通过抑制其转录后翻译负向调节其表达,从而利于胃癌细胞的侵袭。
AIM: To investigate the regulatory effect of miR-193b on the expression of plasminogen activator (uPA) predicted by target gene in gastric cancer cells and its mechanism. Methods: BGC823 cells were transiently transfected with miR-193b sense sequence (sense sequence transfection group), antisense sequence (antisense sequence transfection group) and nonsense sequence (negative transfection group) Cells were blank control. The expression of miR-193b in each group was detected by real-time PCR. The expression of uPA mRNA and protein in each group was detected by RT-PCR and Western blot. Results: Compared with the blank control group, miR-193b expression was not significantly changed in the negative transfected group (P> 0.05), miR-193b expression was significantly up-regulated in the sense sequence transfected group, miR- (All P <0.05). There was no significant change in uPA mRNA level in all transfection groups, uPA protein level in negative transfection group was not changed, uPA protein level in sense transfection group was decreased, The expression of uPA in the transfected group increased. Conclusion: uPA may be the target gene of miR-193b, and miR-193b may negatively regulate its expression by inhibiting its posttranslational transactivation, which will benefit the invasion of gastric cancer cells.