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1. 重庆医科大学医学检验系临床检验诊断学、省部共建教育部重点实验室、重庆市重点实验室
2. 重庆医科大学附属第一医院泌尿外科
纸质出版日期:2009,
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[1]荀春华,罗春丽,吴小候,谢建红,杨淑哲,潘翠翠.hepaCAM基因转染对肾癌786-0细胞增殖和侵袭活性的影响[J].重庆医科大学学报,2009,34(07):841-844.
XUN Chun-hua, et al. Effects of hepaCAM gene transfection on proliferation and invasion of human renal cancer celI line 786-0[J]. Journal of Chongqing Medical University, 2009,34(7):841-844.
目的:肝细胞粘附分子(Hepatocyte cell adhesion molecule
hepaCAM)是近年来发现的一类免疫球蛋白超家族细胞黏附分子
具有抑癌基因特性。本文研究hepaCAM基因转染对肾癌(Renal cell carcinoma
RCC)细胞786-0增殖和侵袭能力的影响。方法:将携有hepaCAM基因的重组质粒hepaCAM-pEGFPN2转染786-0细胞(实验组)后用G418筛选形成阳性克隆并扩增培养
以转染了pEGFPN2(空载组)和未转染的786-0细胞(空白组)为对照
采用逆转录—聚合酶链反应(RT-PCR)方法检测hepaCAMmRNA的表达
四甲基偶氮唑蓝比色法(MTT法)检测786-0细胞增殖活性
Transwell小室法检测细胞体外侵袭能力。结果:稳定转染hepaCAM后786-0细胞的hepaCAMmRNA表达水平明显上调(P<0.01)
MTT显示实验组与空载组相比hep-aCAM基因抑制细胞成活率在4、5、6d分别为26.5%、38.1%、35.7%(P<0.01);侵袭实验显示细胞穿透ECM膜的数目为24.6±5.3(实验组)、35.5±6.23(空载组)和37.1±7.0(空白组)
实验组细胞明显少于2对照组(P<0.01)。结论:抑癌基因hepaCAM转染能降低肾癌细胞786-0的增殖和侵袭活性。
Objective:hepaCAM (hepatocyte cell adhesion molecule)is a kind of immunoglobulin superfamily member which was discovered as a tumor suppressor. This study was to investigate the effects of hepaCAM transfection on proliferation and invasion of human renal cancer cell line 786-0. Methods:An eukaryotic expression plasmid containing hepaCAM
hepaCAM-pEGFPN2 was transfected into 786-0 cells
then positive cell clones were selected with G418 and amplified. pEGFPN2 transfected cells and untreated cells were set as controls.The expression of hepaCAM was detected by reverse transcription-polymerase chain reaction (RT-PCR). Proliferative ability of 786-0 cells was measured by MTT assay and the invasive ability in vitro was determined by transwell chambers method. Results: Plasmid pEGFPN2
hepaCAM-pEGFPN2 were successfully constructed and stably transfected into 786-0 cells.After transfection
the inhibitory rates of cell growth at the fourth、 fifth and sixth day were 26.5%
38.1% and 35.7%
respectively. The penetrating cells were significantly less in 786-0/ hepaCAM-pEGFPN2 group than in 786-0/pEGFPN2 and 786-0 groups (24.6±5.3vs35.5±6.2 and 37.1±7.0
P<0.01). Conclusion:Transfection with hepaCAM might suppress proliferative and invasive ability of renal cancer 786-0 cells.
hepaCAM基因转染肾癌细胞增殖与侵袭
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