【摘要】 目的探讨人端粒酶催化亚基(humantelomerasereversetranscriptase,hTERT)启动子联合mad1基因对膀胱癌细胞T24和EJ细胞的生长抑制作用及机制。方法采用细胞计数、电子显微镜、流式细胞术、RT-PCR及免疫组化等方法,观察hTERT启动子联合mad1基因表达质粒转染T24和EJ细胞后的膀胱肿瘤细胞的增殖能力、生长周期、细胞超微结构的变化以及hTERT和TGF-β1mRNA基因和蛋白表达影响。结果转染hTERT启动子联合mad1基因后对T24和EJ细胞生长有明显抑制作用、出现凋亡小体和细胞周期相的改变,S期细胞明显减少,G0/G1期细胞明显增多,下调T24和EJ细胞hTERT和TGF-β1的mRNA表达,并能抑制hTERT和TGF-β1蛋白表达。结论hTERT启动子联合mad1基因能够抑制膀胱肿瘤细胞的增殖能力,可望为膀胱癌基因治疗提供新思路。更多还原

【Abstract】 Objective To explore the mechanism of the growth inhibition of bladder cancer cells by combination of human telomerase reverse transcriptase （hTERT） promoter and mad1 gene. Methods The cell proliferation, cell cycle progression, cell ultrastructure alterations, mRNA and protein expression of hTERT and TGF-β1 were detected by cell counting, transmission electron microscopy （TEM）, flow cytometry （FCM）, reverse transcription polymerase chain reaction （RT-PCR） and immunocytochemistry after T24 and EJ cells were respectively treated with hTERT promoter and mad1 gene plasmids. Results Bladder cancer cell growth were suppressed, apoptotic bodies were observed, and the cell number of S phase decreased while the cells of G₀/G₁ increased, both mRNA and protein expression of hTERT and TGF-β1 was decreased after hTERT promoter and mad1 gene transfected into T24 and EJ cells. Conclusion Combination of hTERT promoter and mad1 gene could inhibit the proliferation of bladder cancer cells, which may provide a new idea for gene therapy of bladder carcinoma.更多还原