TMSG-1基因转染对肿瘤转移表型的影响  

The effects of TMSG-1 gene transfection on metastatic phenotype of pg cancer cells

作　　者：边巴[1] 马春树[1] 由江峰[1] 宁钧宇[1] 方伟岗[1] 郑杰[1] 

机构地区：[1]北京大学基础医学院病理学系,北京100083

出　　处：《北京大学学报（医学版）》2003年第1期18-22,共5页Journal of Peking University:Health Sciences

基　　金：国家自然科学基金 (3 0 170 3 63 );教育部科学技术研究重大项目 (0 10 0 3 );国家重点基础研究发展规划项目 (973 )(2 0 0 2CB5 13 10 0 );国家"十五"科技攻关项目 (2 0 0 1BA70 3B0 5 )资助~~

摘　　要：目的 :研究肿瘤转移抑制基因TMSG 1对肿瘤转移表型的影响。方法 :将含TMSG 1全长开放阅读框架的 1.5kbcDNA克隆于 pcDNA3,构建正义及反义TMSG 1cDNA真核表达载体 ,以脂质体转染人肺巨细胞癌PG的高转移亚系BE1,挑选G4 18抗性克隆 ,RT PCR检测正义或反义TMSG 1cDNA在转染细胞中的表达 ,进行转染细胞体外肿瘤生物学行为检测。结果 :RT PCR显示正义TMSG 1cDNA转染的BE1细胞 (BE1 S)中TMSG 1表达明显高于BE1及空载体对照细胞 ,反义TMSG 1cDNA转染细胞 (BE1 AS)中TMSG 1表达低于空载体对照细胞。与BE1及空载体对照细胞 (BE1 V)相比 ,BE1 AS细胞体外生长较快 ,软琼脂克隆形成能力明显增强 ;而BE1 S细胞体外生长能力没有显著改变 ,但其穿越Matrigel的能力及软琼脂克隆形成能力明显减弱。流式细胞仪分析结果显示BE1 S的G0 、G1期的细胞较BE1 V细胞比例增多 ,并且BE1 S出现了细胞凋亡峰。结论 :实验结果表明反义TMSG 1基因转染可增强BE1细胞的体外生长、体外侵袭能力及克隆形成能力。而正义TMSG 1基因转染则使BE1细胞的侵袭能力及软琼脂克隆形成能力减弱 ,而对细胞体外生长能力无明显影响。结论支持TMSGObjective: To observe the relationship between TMSG 1 gene and tumor metastatic phenotype. Methods: TMSG 1 cDNA fragment which contained full lenth open reading frame of TMSG 1 gene was cloned into pcDNA3 plasmid to reconstruct sense and antisense eukaryotic expression plasmids of TMSG 1 gene containing neo selection marker. Both sense and antisense eukaryotic expression plasmids of TMSG 1 gene were transfected into the highly metastatic subclone PG BE1 by LipofectAMINE method and the positive clones were selected by G418. RT PCR was used to examine the expression level of the transfected gene and the changes of biological characteristics were checked by a series of in vitro and in vivo assays. Results: The results showed that higher expression levels of TMSG 1 gene in BE1 S cells (cells transfected by sense TMSG 1 cDNA) than the control BE1 cells and BE1 V cells (cells transfected by pcDNA3 plasmid). The expression levels of TMSG 1 gene in BE1 AS cells (cells transfected by antisense TMSG 1 cDNA) were lower than those of the control BE1 cells and BE1 V cells. Compared with the control BE1 cells and BE1 V cells, BE1 AS cells grew more rapidly, and produced more foci in soft agar. Although the BE1 S cells did not reveal significant different growth capacity, the infiltrating ability and colony formation potential of BE1 S cells were decreased, compared with the control BE1 cells and BE1 V cells. Flow cytometry showed higher percentage of BE1 S cells in G 0G 1 phase than that of BE1 cells, and the presence of apoptotic peak in BE1 S cells. Conclusion: The results suggest TMSG 1 gene may represent a tumor metastasis suppressor gene.

关 键 词：TMSG-1基因转染 肿瘤转移 表型 病理生理学 

分 类 号：R730.23[医药卫生—肿瘤]