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 recomhined 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 hy RT - PCR and Western blot. The cell cycle progression and apoptosis were assessed hy flow cytometry. Results A gastric c:ancer 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 GO/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.
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