胃癌相关基因GCRG213真核表达载体的构建及其对胃癌细胞MKN45的影响  

Construction of gastric cancer related gene GCRG213 eukaryotic expression vector and it's effect on gastric cancer cells

作　　者：高利利[1] 王孟薇[1] 吴本俨[1] 王珊[1] 杨怡[2] 黄海力[1] 伍银桥[1] 尤纬缔[1] 王卫华[1] 

机构地区：[1]中国人民解放军总医院,北京100853 [2]军事医学科学院实验仪器中心

出　　处：《山东医药》2008年第41期1-4,共4页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(20370635)

摘　　要：目的研究胃癌相关基因GCRG213正义、反义转染对胃癌细胞MKN45的影响。方法采用分子克隆及基因转染技术将GCRG213基因转入哺乳动物细胞,并结合反义转染技术研究GCRG213基因对胃癌细胞恶性生物学行为的影响。结果GCRG213正向和反向克隆正确插入真核表达载体pcDNA3.1(+);重组子pcDNA3.1-a、pcDNA3.1-b和空载体转染人胃癌细胞系MKN45细胞;正义转染其mRNA的表达上调,而反义转染下调;正义转染加快MKN45细胞生长增殖速度、降低细胞凋亡率,反义转染减慢MKN45细胞生长增殖速度,增加细胞凋亡率。结论胃癌相关基因GCRG213可促进肿瘤细胞生长、分裂和转移,抑制肿瘤细胞的凋亡,可能是一个新发现的恶性肿瘤癌变的促进因素。Objective To investigate the effect of gene GCRG213 transfection （sense, antisense） on gastric cancer cell MKN45. Methods The sense and anti-sense fragment of GCRG213 were cloned into eukaryotic expression vector pcDNA3.1 （ ＋ ） and transfected into MKN45 cells. Results Though sequencing, sense GCRG213 and anti-sense GCRG213 were proved to be successfully cloned into eukaryotic expression vector. The recombinant plasmid pcDNA3, 1-a, pcDNA3, 1-b, and the vector were transfected into MKN45 cells. The MKN45 transfectd by the sense vector significantly increased the expression of GCRG213 , both in mRNA level and protein level. The MKN45 transfected by the anti-sense vector significantly decreased the expression of GCRG213 , both in mRNA level and protein level. The growth of pcDNA3. 1-a transfected cells was faster and the cell apoptosis decreased. But the growth of pcDNA3.1-b transfected cells was slower and the cell apoptosis increased. Conclusion Stable transfection showed that GCRG213 promoted cell growth, malignancy and metastasis, and inhibited the cells into apoptosis. GCRG213 might be a new promoter to tumor.

关 键 词：胃癌相关基因GCRG213 真核表达 胃癌细胞 基因转染 

分 类 号：R735.2[医药卫生—肿瘤]