摘要
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。
Objective： To investigate the regulatory effect of miK-193b on urokinase-type plasminogen activator （uPA）, which was selected according to target gene predictions, in stomach cancer cells and the mechanism. Methods： Gastric cancer BGC823 cells were transiently transfected with the miK-193b sense sequence （sense sequence transfection group）, miR-193b antisense sequence （antisense sequence transfection group） and scrambled sequence （negative transfection group） respectively, with the untransfected BGC823 cells as a blank control. In each group of cells, the expression levels of miR-193b were detected by real-time PCR, and themRNA and protein expressions were determined by RT-PCR and Western blot analysis, respectively. Results： Compared with blank control group, the miR-193b expression in negative transfection group had no obvious change （P〉0.05）, while it was significantly increased in sense sequence transfection group and significantly decreased in antisense sequence transfection group （both P〈0.05）; the uPA mRNA expression levels in all transfection groups had no obvious change; the uPA protein expression in negative transfection group showed no apparent change, but it was reduced in sense sequence transfection group and increased in antisense sequence transfection group. Conclusion： uPA may be the target gene of miR-193b; miR-193b may negatively regulate its expression through inhibition of its post-transcriptional translation, and thereby enhance the invasiveness of gastric cancer cells.
出处
《中国普通外科杂志》
CAS
CSCD
北大核心
2013年第12期1585-1589,共5页
China Journal of General Surgery
