Objective To investigate the effects of ALKBH5 on migration, invasion and epithelial-mesenchymal transformation (EMT) of hepatoma cells.
Methods A total of 50 hepatocellular carcinoma specimens were selected. The relationship between ALKBH5 and clinicopathological parameters was analyzed. ALKBH5 short hairpin RNA (shRNA) sequence was designed, pLKO.1-TRC vector was constructed, then lentivirus was used to infect HepG2 and SMMC7721 cells. Puromycin screened and cultured ALKBH5 cell lines with stable low expression. Interference efficiency of ALKBH5 mRNA and ALKBH5 protein levels were detected. Transwell assay was used to detect the migration and invasion of cells. Western blot was used to detect the effects of EMT-associated proteins.
Results ALKBH5 protein expression was correlated with differentiation degree and clinical stage (P < 0.05), but not with serum alpha-fetoprotein (AFP) level, tumor size, age and gender (P>0.05). The expression of ALKBH5 was down-regulated, and the migration number of HepG2 cells in control group and experimental group was (123±22) and (335±21), respectively; the number of invasions was (118±29) and (282±17), respectively; the migration number of SMMC7721 cells in control group and experimental group was (220±27) and (453±23), respectively; the number of invasions was (200±11) and (388±20), respectively, and the differences among all groups were statistically significant (P < 0.05). After down-regulated ALKBH5 expression, E-cadherin protein expression was decreased, and the expression level of Vimentin and Snail protein increased (P < 0.05).
Conclusion ALKBH5 plays the role of tumor suppressor gene, and down-regulated ALKBH5 expression can promote migration, invasion and EMT of hepatoma carcinoma cells.
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