山东医药Issue(10)：1-3,3.DOI:10.3969/j.issn.1002-266X.2014.10.001
高表达重组人组织激肽释放酶7基因的前列腺癌单克隆细胞株的构建
Establishment of a human prostatic carcinoma cell line stably overexpressing Kallikrein 7
摘要
Abstract
Objective To establish a human prostatic carcinoma cell line stably overexpressing Kallikrein 7 (KLK7) and provide foundation for prostate cancer pathogenesis .Methods KLK7 cDNA was amplified by PCR and cloned into the eukaryotic expression vector pcDNA3.1.Then it was confirmed by restriction endonuclease and DNA sequencing test , and the correct vectors were transfected into prostate cell line DU 145 with lipofectamine .The survival cell clones treated by G418 were amplified.The KLK7 expression of cell line DU145 was detected by Western blotting in each cell clone .Re-sults After restriction endonuclease analysis and DNA sequencing , the pcDNA3.1-KLK7 eukaryotic expression vector was successfully constructed .The stably transfected DU145 cell line highly expressing KLK7 was successfully established after being transfected .The results of Western blotting showed the expression of KLK 7 protein was significantly higher in stable transfectants with the KLK7 vector than that in the cells with empty vector (P<0.01).Conclusion The construc-tion of the eukaryotic expression vector pcDNA 3.1-KLK7 and the establishment of stable prostatic cell line overexpressing KLK7 provide basis for further studies on the function KLK 7 in the prostate pathogenesis .关键词
重组人组织激肽释放酶基因7/前列腺癌/稳定细胞株/真核表达载体Key words
recombinant human kallikrein 7/prostate carcinoma/stable cell line/eukaryotic expression vector分类
医药卫生引用本文复制引用
林志弟,莫林键,张成东,宣强,张悦宁,莫曾南,滕若冰,杨小丽..高表达重组人组织激肽释放酶7基因的前列腺癌单克隆细胞株的构建[J].山东医药,2014,(10):1-3,3.基金项目
国家自然科学基金资助项目（81060214）；高等学校博士学科点专项科研基金资助课题（20104503120008）；广西壮族自治区自然科学基金资助项目（2011GXNSFA018175）。 （）
