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该研究探讨了6-甲基腺嘌呤(N6-methyladenosine,m~6A)去甲基酶ALKBH5(alk B homolog5)对人胃癌AGS细胞迁移和侵袭的影响。通过数据库分析ALKBH5在胃癌组织中的表达水平;Real-time PCR和Western blot检测ALKBH5在胃癌细胞中表达水平;用sh-EGFP和sh-ALKBH5质粒转染人胃腺癌AGS细胞,Transwell实验检测细胞的迁移和侵袭能力,细胞划痕实验进一步检测细胞的迁移能力;用Western blot检测上皮–间质转化(epithelial-mesenchymal transition,EMT)相关蛋白质的变化。结果显示,弥漫性胃腺癌中ALKBH5的m RNA水平明显低于正常胃组织和其他类型的胃癌组织,且其基因拷贝数也明显低于正常胃组织。在胃癌细胞系中,AGS细胞ALKBH5蛋白质和m RNA水平最高,sh-ALKBH5干扰能明显促进AGS细胞迁移和侵袭能力,且下调其上皮指标水平,而上调间质指标水平。上述结果提示,ALKBH5在胃癌组织中可能是一个抑癌基因,与胃癌细胞的迁移和侵袭能力负相关。
This study investigated the effects of alkylating homolog 5-alkylase (N6-methyladenosine, m ~ 6A) on the migration and invasion of human gastric cancer AGS cells. The expression of ALKBH5 in gastric cancer tissues was analyzed by real-time PCR and Western blot. The expression of ALKBH5 in gastric cancer cells was detected by Real-time PCR and sh-EGFP and sh-EGFP and sh-ALKBH5, respectively. Migration and invasion ability. Cell scratch assay was used to further detect the migration of cells. Western blot was used to detect the changes of proteins related to epithelial-mesenchymal transition (EMT). The results showed that the m RNA level of ALKBH5 in diffuse gastric adenocarcinoma was significantly lower than that in normal gastric tissues and other types of gastric cancer tissues and the gene copy number was also significantly lower than that in normal gastric tissues. In gastric cancer cell lines, ALKH5 protein and m RNA levels were the highest in AGS cells. The sh-ALKBH5 interference could significantly promote the migration and invasion of AGS cells, down-regulate the level of epithelial index and up-regulate the level of interstitial markers. The above results suggest that ALKBH5 may be a tumor suppressor gene in gastric cancer tissues and negatively correlated with the ability of gastric cancer cells to migrate and invade.