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1. 重庆医科大学附属第一医院泌尿外科
2. 重庆医科大学检验系
3. 重庆医科大学
纸质出版日期:2009,
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[1]杨超,吴小候,罗春丽,蔡晓钟,吴绮思.HepaCAM基因对人膀胱癌细胞增殖的抑制作用研究[J].重庆医科大学学报,2009,34(09):1157-1161.
YANG Chao, et al. The inhibitory effect of gene HepaCAM transfection on the proliferation in human bladder carcinoma cell line[J]. Journal of Chongqing Medical University, 2009,34(9):1157-1161.
目的:研究hepaCAM基因对人膀胱癌T24细胞增殖的影响。方法:运用脂质体转染方法
将pEGFP-N2-hepaCAM质粒转入T24细胞株
经G418筛选获得稳定表达的细胞株;RT-PCR及Westernblot方法检测目的基因和蛋白表达;MTT法及克隆形成实验研究hepaCAM基因对T24细胞体外生长的作用;流式细胞仪检测细胞周期分布;RT-PCR技术测定周期调控蛋白CyclinD1的mRNA表达变化。结果:RT-PCR和Westernblot方法分别检测到hepaCAM的基因表达和蛋白表达
MTT检测转染hepaCAM基因的细胞生长速度较对照组和空载体组明显减慢(P<0.01)
克隆形成率明显小于对照组和空载体组(P<0.01)。细胞周期中G0/G1期比例明显增高(P<0.05)
而S期比例减少;CyclinD1 mRNA表达明显下调(P<0.05)。结论:HepaCAM基因通过阻滞细胞于G0/G1期
减少CyclinD1 mRNA表达
抑制T24细胞的增殖。
Objective:To explore the effect of gene hepaCAM transfection on the proliferation in human bladder carcinoma cell line. Methods: Plasmid pEGFP-N2-hepaCAM was transfected into T24 cells by Lipofectamine2000
and the T24 cell stably expressing hepaCAM gene was established by G4l8 selection.The expression of hepaCAM mRNA and protein was detected by RT-PCR and Western blot assays. The effects of hepaCAM on cell proliferation were measured by MTT and colony formation assays. The cell cycle progression were measured by flow cytometry. The expression of CyclinD1 mRNA was detected by RT-PCR. Results:Expression of hepaCAM gene and protein were proved by RT-PCR and Western blot assays. The MTT assay showed cell growth in the T24/hepaCAM-GFP group was significantly slow when compared with non-transfection and mock-vehicle groups whereas the rates of clonal formation were greatly decreased(P<0.01). Flow cytometric data showed that more positive clone cells were blocked into G0/G1 phase and fewer cells went into phase S(P<0.05). The expression of CyclinD1 mRNA was lower than non-transfection and mock-vehicle groups. Conclusion: HepaCAM gene might suppress the proliferation of T24 cells through arresting cell cycle in the G0/G1 phase and inhibitting the expression of CyclinD1 mRNA.
膀胱癌hepaCAM基因T24细胞转染抑癌基因
Transitional cell carcinoma of bladderHepaCAMT24 cellTransfactiontumor suppressor gene
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