Liu Jing, Li Li-Jun, Qiu Ming-Xing
Department of Urology, Sichuan Academy of Medical Sciences,Sichuan Provincial People's Hospital,Chengdu 610072,China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2016 Sep;47(5):665-668.
To examine the effects of on cell proliferation and apoptosis and the expression level of Smad4 mRNA in the bladder cancer cell line of T24 by transfection with recombinant plasmid of pIRES-.
The recombinant plasmid of pIRES- was constructed successfully. Cultured T24 cells were divided into three groups, including control group, empty vector group,and recombinant plasmid group. The cells in empty vector group and recombinant plasmid group were respectively transfected by pIRES- and pIRES- The cells were harvested at 24 h after the transfection, the variation of cell morphology was examined by fluorescence microscopy. The cell apoptosis was detected by flow cytometry. The expression level of and Smad4 mRNA was measured by RT-PCR.
Cell death was observed in two transfection groups. At 24 h after transfection,the apoptosis rate was (3.23±0.45)% in control group, (8.98±1.62)% in empty vector group and (43.61±2.69)% in recombinant plasmid group. The expression level of mRNA was 2.79±0.36,detected only in recombinant plasmid group, which was significantly up-regulated compared with the other two groups (<0.05).
The expression level of Smad4 mRNA was up-regulated by transfection with pIRES-,which also inhibited cell proliferation and promoted cell apoptosis.The tumor suppressor gene of could regulate the bladder cancer cell proliferation and apoptosis by TGF-β/Smad signaling pathway.
通过转染重组质粒pIRES-，研究其对膀胱癌细胞系T24细胞增殖、凋亡及Smad4 mRNA表达水平的影响。
成功构建重组质粒pIRES-。将培养的T24细胞分为三组，即对照组、空载体组和重组质粒组。空载体组和重组质粒组细胞分别用pIRES-和pIRES-转染。转染后24 h收获细胞，用荧光显微镜观察细胞形态变化。采用流式细胞术检测细胞凋亡。用RT-PCR检测和Smad4 mRNA的表达水平。
两个转染组均观察到细胞死亡。转染后24 h，对照组凋亡率为(3.23±0.45)%，空载体组为(8.98±1.62)%，重组质粒组为(43.61±2.69)%。mRNA表达水平为2.79±0.36，仅在重组质粒组检测到，与其他两组相比显著上调(<0.05)。
转染pIRES-可上调Smad4 mRNA表达水平，同时抑制细胞增殖并促进细胞凋亡。的抑癌基因可通过TGF-β/Smad信号通路调节膀胱癌细胞的增殖和凋亡。
