摘要
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 MTF 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-O/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.
出处
《重庆医科大学学报》
CAS
CSCD
北大核心
2009年第7期841-844,共4页
Journal of Chongqing Medical University
