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Effects of ALKBH5 on the Abilities of Migration and Invasion in Human Gastric Cancer Cell Line AGS
Lu Ying1, Wang Dawei1, He Junbo1, Zhou Meng1, Zeng Jian2, Gong Aihua2, Xu Min1*
1Department of Gastroenterology, Affiliated Hospital of Jiangsu University, Zhenjiang 212013, China; 2School of Medicine, Jiangsu University, Zhenjiang 212013, China
Abstract: The aim of this work is to investigate the effects of m6A (N6-methyladenosine) demethylase ALKBH5 (alkB homolog 5) on the abilities of migration and invasion in human gastric cancer cell line AGS. The levels of ALKBH5 gene expression in gastric cancer tissues were explored in databases. Real-time PCR and Western blot were used to detect the relative levels of mRNA and protein of ALKBH5 in human gastric cancer cell lines， respectively. Then plasmids sh-EGFP and sh-ALKBH5 were transfected into AGS cells. Transwell assays were used to examine the abilities of cell migration and invasion. The migration rate was measured by wound healing assay. Epithelial-mesenchymal transition (EMT) markers were detected by Western blot. Compared with normal gastric tissue， mRNA levels and gene copy numbers of ALKBH5 were decreased in high invasiveness diffuse gastric adenocarcinoma. The relative levels of ALKBH5 protein and mRNA were differed in different gastric cancer cells， and were highest in AGS. The down-regulation of ALKBH5 could enhance the abilities of migration and invasion in AGS cells. The relative level of epithelial marker was decreased， and mesenchymal markers were increased in ALKBH5 down-regulated AGS. The results suggested that ALKBH5 might be a tumor suppressor gene and related to the migration and invasion abilities of human gastric cancer cell.
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