Intracellular and cellular functions of ALKBH5 in thyroid cancer cells
OBJECTIVE:To investigate the role of N6-methyladenosine(m6A)demethylase ALKBH5 on intracellular and cellular functions of thyroid carcinoma cells in vitro.METHODS:The ALKBH5 knockdown plasmid was transfected to human thyroid papillary carcinoma cells(TPC-1)and undifferentiated thyroid carcinoma cells(8505C).Expression of m6A in the cancer cells were measured using a m6A methylation quantification detection kit.Plate cloning,CCK-8,plate scratch,and Transwell were used to analyze the proliferation,migration,and invasion ability.Flow cytometry was used to detect the cell cycle of the cancer cells and the percentage of cell in each phase.The relationship between ALKBH5 and downstream pathways was analyzed by Western blot experiments.RESULTS:Compared to cells without the ALKBH5 knockdown,the knockdown increased m6A expression of levels in the TPC-1 and 8505C cancer cells,and decreased the number of cell proliferation,invasion,and migration decreased(P＜0.05).In addition,cells were arrested at the G2/M phase of cell cycle.Western blot experiments showed that when the expression of ALKBH5 was low,the key genes of the PI3K/AKT signaling pathway were perturbed.Protein expressions of P85,AK,P-AKT and FAK were inhibited,as well as that of CDK4,CDK6,Cyclin B1,Cyclin D1,Cyclin E1 in the downstream Cyclin pathway,but P53 was increased(P＜0.05).CONCLUSION:In the thyroid papillary carcinoma cells,ALKBH5 knockdown inhibited their proliferation,migration,invasion and cell cycle,as well as the PI3K/AKT/P53 and PI3K/AKT/Cyclin axis.
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