N-乙酰氨基葡萄糖转移酶Ⅴ在CD133^+人肺腺癌细胞中的表达和功能研究  

Expression and Function of Mgat5 in CD133^+ Pulmonary Adenocarcinoma Cells

作　　者：周雪峰[1] 王乐[1] 张力[1] 但攀[1] 赵金平[1] 

机构地区：[1]武汉大学中南医院胸心外科,湖北武汉430071

出　　处：《武汉大学学报（医学版）》2011年第5期589-592,共4页Medical Journal of Wuhan University

基　　金：国家自然科学基金资助项目(编号:81101775);中央部属高校专项科研基金资助项目(编号:111118);湖北省卫生厅科研项目(编号:NX2011-6)

摘　　要：目的:观察N-乙酰氨基葡萄糖转移酶Ⅴ(Mgat5)在CD133+人类肺腺癌细胞中的表达和功能。方法:采用磁珠分选(MACS)的方法从10位肺腺癌患者手术切除的肿瘤标本中分离出CD133+人类肺腺癌细胞。荧光实时定量PCR(FQRT-PCR)和Western blot检测CD133+细胞中Mgat5的表达。Mgat5特异性的siRNA转染CD133+细胞后,L型植物凝集素(L-PHA)结合实验检测细胞表面N-聚糖表达情况。体内荷瘤实验和MTT比色法分别检测敲减Mgat5后CD133+肺腺癌细胞体内外的生长情况。结果:CD133+细胞中Mgat5的mRNA和蛋白表达量分别是CD133-细胞中Mgat5的1.2倍和1.4倍。CD133+细胞转染Mgat5特异性siRNA后与L-PHA结合能力下降,敲减CD133+细胞中Mgat5的表达能够显著抑制肿瘤细胞体内和体外的生长。结论:Mgat5在CD133+细胞内高表达,抑制Mgat5的表达可明显抑制肿瘤细胞的生长。Mgat5可以作为一个新的肺腺癌治疗靶点。Objective：To investigate the expression and function of N-acetylglucosaminyltransferase Ⅴ（Mgat5） in CD133＋ pulmonary adenocarcinoma cells.Methods：CD133＋ pulmonary adenocarcinoma cells were separated by MACS from excised pulmonary adenocarcinoma specimens from 10 patients.Expression of Mgat5 in CD133＋ cells was detected by Fluorescence Quantitative RT-PCR（FQRT-PCR） and Western blot.Subsequently,CD133＋ cells were transfected with specific siRNA of Mgat5 to evaluate the inhibition of Mgat5 on cancer cell growth in vivo and in vitro.Results：Expression of Mgat5 were 1.2 folds and 1.4 folds higher in CD133＋cells than in CD133– cells detected by FQRT-PCR and Western blot respectively.The L-PHA binding assay also showed higher reactivity in CD133＋ cells than in CD133-cells.Interestingly,downregulation of Mgat5 by Mgat5 specific siRNA resulted in significant inhibition of cancer cell growth in vitro and in vivo.Conclusion：Mgat5 may play an important role during oncogenesis,and it is a potential therapeutic target for pulmonary adenocarcinoma.

关 键 词：Mgat5 CD133 肺腺癌 N-聚糖 

分 类 号：R734.2[医药卫生—肿瘤]