稳定过表达Cdx2基因胃癌细胞株的建立  

Establishment of gastric cancer cell line with stable over-expression of Cdx2

作　　者：李雷[1] 王晓通[1] 谢玉波[2] 肖强[1] 

机构地区：[1]广西医科大学第一附属医院胃肠腺体外科,南宁530021 [2]广西医科大学第一附属医院麻醉科,南宁530021

出　　处：《广东医学》2011年第8期971-973,共3页Guangdong Medical Journal

基　　金：国家自然科学基金资助项目(编号:81060201);中国博士后科学基金资助(编号:201003342)

摘　　要：目的建立稳定过表达Cdx2基因的胃癌细胞株。方法使用阳离子脂质体分别将真核表达载体pCMV-Cdx2-HA或空载体pCMV-HA转染至人胃癌细胞MGC-803中,G418筛选出阳性克隆后扩大培养。RT-PCR和Western Blot技术检测各组胃癌细胞中Cdx2基因mRNA和蛋白的表达情况,将挑选出的稳定株命名为MGC-803/Cdx2细胞(转染组)和MGC-803/EV细胞(空载体组);流式细胞仪检测MGC-803细胞(未转染组)、MGC-803/EV细胞和MGC-803/Cdx2细胞的细胞周期和凋亡情况。结果成功筛选出稳定过表达Cdx2基因的MGC-803胃癌细胞,即MGC-803/Cdx2细胞;与MGC-803细胞和MGC-803/EV细胞比较,MGC-803/Cdx2细胞中Cdx2基因mRNA和蛋白的表达明显增加(P<0.05),而且细胞周期G0/G1期比例和凋亡率也明显增加(P<0.05)。结论成功构建了稳定过表达Cdx2基因的胃癌细胞株,而且Cdx2过表达使细胞周期停滞、凋亡增加。Objective To establish a gastric cancer cell line with stable over - expression of Cdx2 gene. Methods The reeombined eukaryotic expression vector pCMV - Cdx2 - HA or empty vector pCMV - HA was transfected into gastric cancer MGC -803 cells by Lipofectamine. G418 -resistant colonies were selected by adding G418 to the medium. Cdx2 mRNA and protein expression was assessed by RT - PCR and Western blot. The cell cycle progression and apoptosis were assessed by flow cytometry. Results A gastric cancer cell line with stable over - expression of Cdx2 was established successfully, which was named MGC-803/Cdx2 cells. Cdx2 mRNA and protein in MGC -803/Cdx2 cells significantly increased, compared with those in MGC - 803 cells and MGC - 803/EV cells （ P 〈 0. 05 ）. The proportion of cells in G0/ G1 phase and the apoptotic rate in MGC - 803/Cdx2 cells were significantly higher than those in MGC - 803 cells and MGC - 803/EV cells （ P 〈 0. 05 ）. Conclusion Gastric cancer cell line with stable over - expression of Cdx2, which induces cell cycle arrest and cell apoptosis, is established.

关 键 词：CDX2 胃癌 细胞周期 凋亡 

分 类 号：R742.5[医药卫生—神经病学与精神病学]