腺病毒介导FOXA2基因过表达对胃癌细胞生长影响

EFFECT OF ADENOVIRUS-MEDIATED FOXA2 OVEREXPRESSION ON THE GROWTH OF GASTRIC CANCER CELLS

目的探讨腺病毒介导的FOXA2基因过表达对胃癌细胞生长的影响及其机制。方法根据人叉头蛋白A2(FOXA2)mRNA的序列,设计合成FOXA2基因的引物,扩增获得全长FOXA2,构建腺病毒表达载体,进一步获得腺病毒Ad5.FOXA2。应用腺病毒Ad5.FOXA2感染胃癌细胞系,实时荧光定量PCR和Western blot方法检测FOXA2基因和蛋白表达情况。应用Western blot、甲基噻唑基四唑(MTT)、细胞划痕实验等方法,检测FOXA2基因对胃癌细胞增殖和迁移的作用。结果成功构建了携带FOXA2的腺病毒Ad5.FOXA2。Ad5.FOXA2感染胃癌细胞系研究显示,Ad5.FOXA2组FOXA2蛋白表达是对照组和Ad5.GFP组的3.85倍;各组FOXA2mRNA表达量比较,差异有统计学意义(F=6.896,P<0.05)。MTT检测显示,Ad5.FOXA2抑制了胃癌细胞的增殖及迁移。Western blot检测显示,Ad5.FOXA2抑制了c-Myc、β-catenin、Cyclin D1以及PCNA蛋白的表达。结论腺病毒Ad5.FOXA2抑制胃癌细胞生长,其作用是通过抑制c-Myc、β-catenin、Cyclin D1和PCNA蛋白的表达实现的。

Objective To investigate the effect of adenovirus mediated FOXA2 overexpression on the growth of gastric cancer cells and the related mechanism. Methods The primer of the FOXA2 gene was designed and synthesized according to the mRNA sequence of FOXA2, and full-length FOXA2 was obtained by multiplication. The adenovirus expression vector was con- structed, and then the adenovirus Ad5.FOXA2 was obtained. Gastric cancer cells were infected by the adenovirus Ad5.FOXA2, and quantitative real-time PCR and Western blot were used to measure the mRNA and protein expression of FOXA2. Western blot, methyl thiazolyl tetrazolium （MTT） assay, and wound healing test were used to evaluate the effect of the FOXA2 gene on the pro- liferation and migration of gastric cancer cells. Results The adenovirus Ad5.FOXA2 which carried FOXA2 was successfully constructed. After the gastric cancer cells were infected by Ad5.FOXA2, the protein expression of FOXA2 in the Ad5. FOXA2 group was 3.85 times that in the control group and the AdS.GFP group, and there was a significant difference in the mRNA expres- sion of FOXA 2 between groups （F ： 6.896, P d0.05）. The MTT assay showed that Ad5. FOXA2 inhibited the proliferation and migration of gastric cancer cells, and Western blot showed that Ad5.FOXA2 inhibited the expression of c-Myc, β-catenin, Cyclin D1, and PCNA. pression of c Myc, Conclusion The adenovirus AdS.FOXA2 inhibits the growth of gastric cancer cells by inhibiting the protein ex β-catenin, Cyclin D1, and PCNA.