稳定过表达Cdx2基因的胃癌细胞株的建立

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。

Establishment of a gastric cancer cell line with stable overexpressed Cdx2 gene

Objective To establish a gastric cancer cell line with stable overexpressed Cdx2 gene. Methods The recombined eukaryotic expression vector pCMV-Cdx2-HA or empty vector pCMV-HA was transfected into gastric cancer MGC-803 cells individually by Lipofectamine. G418-resistant colonies were selected by adding G418 to the medium. Cdx2 mRNA and protein expression were detected by RT-PCR and Western blotting. The cell cycle progression and apoptosis were assesed by flow cytometry. Results A gastric cancer cell line with stable overexpressed Cdx2 gene was established successfully, which is named as MGC-803/Cdx2 cells. Cdx2 mRNA and protein in MGC-803/Cdx2 cells were increased compared with 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 higher than that in MGC-803 cells and MGC-803/EV cells (P<0.05). Conclusions A gastric cancer cell line with stable overexpressed Cdx2 gene was established and Cdx2 overexpression made cell cycle arrest and promoted cell apoptosis.