MicroRNA-1243 regulates cell migration by targeting serine/threonine protein kinase 1 and serine/threonine protein kinase 2 in thyroid papillary carcinoma TPC-1 cell

Abstract

Objective To study microRNA (miRNA, miR)-1243 direct targeting serine/threonine protein kinase 1 (Akt1) and serine/threonine protein kinase 2 (Akt2) regulation of thyroid papillary carcinoma TPC-1 cell migration. Methods miR-1243 target gene was predicted by TargetScan and miRanda. The double luciferase reporter gene was used to verify the direct regulation of miR-1243 on target gene. Western blotting was used to detect the protein level of Akt1 and Akt2 in miR-1243 transfected cell. The migration of cells was detected by transwell. Results The TargetScan and miRanda software predict that the Akt1 and Akt2 genes are potential target genes of miR-1243. The relative fluorescence values of the double fluorescent reporter gene showed that the activity of luciferase in the Akt1 or Akt2 wild-type group transfected with miR-1243 mimics was significantly lower than that of the empty vector group and the mutant group (t=3.595, P=0.018). There was no significant difference in luciferase activity between the vector group and the mutant group (t=1.655, P=0.213). Compared with the control group, the miR-1243 mimic treatment group was significantly inhibited and the miR-1243 inhibitor treatment group promoted the expression of Akt1 and Akt2 protein in TPC-1 cells (t=4.810, P=0.009), but the protein level of the internal reference gene was not changed (P=0.235). The results of transwell showed that the number of migratory cells in the miR-1243 mimic transfected group [(9.83±3.51) cells] was lower than that in the control group [(18.67±4.24) cells], the difference was statistically significant (t=2.692, P=0.039). Compared with the control inhibitor group [(21.33±5.35) cells], the number of migrating cells in the miR-1243 inhibitor transfection group [(37.17±7.33) cells] was significantly increased (t=2.472, P=0.022). Conclusion miR-1243 can inhibit the expression of Akt1 and Akt2 and regulate the migration of TPC-1 cells. Key words: TPC-1 cells; MicroRNA-1243; Cell migration; Serine/threonine kinase 1; Serine/threonine kinase 2