nm23-H1基因在人肺癌细胞L9981恶性表型逆转中的作用  

Effect of nm23-H1 on Reversing Malignant Phenotype on Human Lung Cancer Cell Line L9981

作　　者：车国卫[1] 周清华[1] 王艳萍[2] 刘伦旭[1] 覃杨[2] 孙芝琳[2] 孙泽芳[2] 陈晓禾[2] 

机构地区：[1]四川大学华西医院胸心外科,成都610041 [2]四川大学华西医院四川省肺癌分子重点实验室,成都610041

出　　处：《生物医学工程学杂志》2005年第3期530-534,共5页Journal of Biomedical Engineering

基　　金：国家自然科学基金资助项目(30070333;30100075);中国博士后科学基金资助(20040350134)

摘　　要：nm23-H1基因是肿瘤转移抑制基因,转染野生型的nm23-H1基因可以逆转肺癌的恶性表型,我们拟建立转染野生型nm23-H1基因的人大细胞肺癌细胞株L9981-nm23-H1,并初步探讨nm23-H1基因在逆转L9981细胞株恶性表型中的作用。应用基因克隆技术构建质粒载体pLXSN-nm23-H1-EGFP,并建立L9981-nm23-H1细胞株。同时应用Westernblot检测L9981-nm23-H1细胞中nm23-H1蛋白表达。细胞培养及改良的Boyden小室法分别检测L9981-nm23-H1细胞株的体外生物学行为,动物实验检测L9981-nm23-H1细胞株在裸鼠体内的成瘤性及肺部转移能力。结果成功构建了逆转录病毒载体pLXSN-nm23-H1-EGFP;并建立了人大细胞肺癌细胞株L9981-nm23-H1;nm23-H1基因在细胞株L9981-nm23-H1中稳定、高效表达;L9981-nm23-H1细胞体外增殖能力,克隆形成力,体外侵袭力显著降(P<0.01);L9981-nm23-H1裸鼠体内的成瘤性及肺部转移能力显著低于L9981和L9981-pLXSN(P<0.01);nm23-H1基因的抑瘤率82.56%。本研究资料提示转染野生型nm23-H1基因可以逆转人大细胞肺癌细胞株L9981的恶性表型。nm23-H1 is a proven tumor metastasis suppressive gene, tumor metastasis phenotype could be reversed by transfected nm23-H1 cDNA. This study was conducted to transfect nm23-H1 cDNA into L9981 cells and to explore the function of nm23-H1 in reversing the malignant phenotype of L9981 cells. The plasmid of pLXSN-nm23-H1-EGFP was constructed by gene clone technique, and the transfected nm23-H1 cDNA cell lines of L9981-nm23-H1 was established. The protein expression of nm23-H1 was detected by Western blot. The biologic features of L9981-nm23-H1 cells were studied in vitro and in vivo. The results showed that the fusion protein of nm23-H1-EGFP was stable, continuous and expressed with high efficiency in L9981-nm23-H1 cells. The cell proliferation, colon formation and invasive ability are significantly lowered in L9981 cells transfected nm23-H1 cDNA （P〈0.01）; the tumorgenesis and the lung metastasis incidence was lower in tranfected nm23-H1 cells than in L9981 and L9981-Plxsn in nude mice （P〈0. 01）; the rate for inhibiting tumorgenesis of nm23 -H1 was 82.56 %. These data suggest that the malignant phenotype could be reversed by wild nm23-H1 gene in L9981 cells.

关 键 词：NM23-H1基因 人肺癌 肿瘤细胞 L9981 恶性表型 逆转录病毒载体 

分 类 号：R734.2[医药卫生—肿瘤]