HepaCAM基因转染对肾癌细胞侵袭活性的影响  

Effects of HepaCAM gene transfection on invasiveness of renal cancer cells

作　　者：荀春华[1] 罗春丽[1] 吴小候[2] 谢建红[1] 杨淑哲[1] 潘翠翠[1] 

机构地区：[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400016 [2]重庆医科大学附属第一医院泌尿外科,重庆400016

出　　处：《肿瘤》2008年第12期1029-1033,共5页Tumor

摘　　要：目的:研究肝细胞黏附分子(hepatocyte cell adhesion molecule,hepaCAM)基因转染对肾细胞癌(renal cell carcinoma,RCC)细胞株786-0侵袭能力的影响,并探讨其作用机制。方法:将携带hepaCAM基因的重组质粒hepaCAM-pEGFPN2转染786-0细胞后,用G418筛选形成阳性克隆并扩增培养,以转染了pEGFPN2和未转染的786-0细胞作为对照。采用RT-PCR和实时荧光定量PCR法检测hepaCAM和基质金属蛋白酶(matrixmetalloproteinase,MMP)-2和MMP-9 mRNA的表达,明胶酶谱法检测MMP-2和MMP-9的活性变化,Transwell小室法检测786-0细胞的体外侵袭能力。结果:稳定转染hepaCAM基因后,786-0细胞的hepaCAM mRNA表达水平明显上调(P<0.01),而MMP-2和MMP-9 mRNA的表达和酶活性明显受抑(P<0.05),hepaCAM基因的导入显著削弱了RCC细胞的跨膜侵袭能力(P<0.01)。结论:hepaCAM可能通过下调MMP-2和MMP-9的表达以及抑制明胶酶的活性,进而抑制肾癌细胞786-0的侵袭力。Objective： This study was to evaluate the effects of hepaCAM （hepatocyte cell adhesion molecule） on invasiveness of human renal cell carcinoma （RCC） cell line 786-0 and explore the underlying mechanism. Methods：A recombinant plasmid containing hepaCAM cDNA （hepaCAM-pEGFPN2） was transfected into 786-0 ceils, then the positive cell clones were selected with G418 and amplified. The 786-0 cells transfected with pEGFPN2 and untreated 786-0 ceils were as controls. The mRNA expressions of hepaCAM and matrix metalloproteinase （MMP） -2 and MMP-9 before and after stable transfection are detected by reverse transcriptase- polymerase chain reaction （RT-PCR） and real-time fluorescence PCR. The activities of MMP-2 and MMP-9 were analyzed by gelatin zymography. The invasiveness of 786-0 cells was determined by Transwell migration assay in vitro. Results： The mRNA transcription of hepaCAM was significantly increased in 786-0 cells after stable transfection of hepaCAM （P 〈 0.01 ）. The mRNA expressions and activities of MMP-2 and MMP-9 in 786-0 cells were markedly inhibited by hepaCAM （P 〈 0. 05 ）. The transmembrane migration capacity of cells was significantly reduced after stable transfection of hepaCAM （ P 〈 0.01 ）. Conclusion： HepaCAM inhibits the activities of gelatinase by down-regulating the expressions of MMP-2 and MMP-9, thereby preventing the invasiveness of renal cell carcinoma 786-0 cells.

关 键 词：肾肿瘤 细胞黏附分子 转染 肿瘤侵袭 肿瘤细胞 培养的 基因 hepaCAM 

分 类 号：R737.11[医药卫生—肿瘤]