Mechanism of microRNA-1243 on proliferation and migration of human papillary thyroid carcinoma TPC-1 cells

Abstract

Objective To observe the biological effects of microRNA (miRNA, miR)-1243 overexpression on proliferation and migration of human papillary thyroid carcinoma cell line TPC-1. Methods The expression of miR-1243 in 2×105 TPC-1 cells was detected by real-time quantitative polymerase chain reaction (RT-qPCR). The capacities of migration and the migration in TPC-1 cells were tested by water soluble methacon compounds (MTS) or Transwell assays, respectively. The candidate targets for miR-1243 were investigated using predication algorithm provided by miRanda and RNAhybrid. Luciferase activity was measured by dual luciferase reporter assay. Results The expression levels of miR-1243 in blank control and negative control groups were 0.97±0.31 and 1.208±0.40 respectively, significantly lower than 932.84±52.34 in miR-1243 mimic transfected group (P=0.000). The relative survival rates of the blank control group and the negative control group were 4.82±0.10 and 4.91±0.09, respectively, which were significantly higher than that of 4.41±0.06 in miR-1243 mimic transfected group (P=0.032). The proliferation ability of TPC-1 cells transfected with miR-1243 was significantly inhibited. The number of migratory cells in the miR-1243 mimic group was 15.5±6.5, which was significantly less than that in the blank control group (33.5±9.5) and the negative control group (31.75±10.75) (P=0.016). The bioinformatics analysis revealed that the AKT serine/threonine kinase 1 (AKT1) mRNA may be the target of miR-1243. The results of dual luciferase assay showed that WT 3’ untranslated region (UTR) of AKT1 cotransfected with miR-1243 mimics significantly decreased luciferase activity (45.92±10.91) as compared with empty vector group (98.97±9.96), mutant group (85.02±11.04) and negative control mimic group (97.38±10.29) (P=0.005). Conclusion Overexpression of miR-1243 can inhibit the proliferation and migration of TPC-1 cells. The miR-1243 may be involved in the regulation of thyroid papillary carcinoma by direct targeting of AKT1. Key words: Thyroid papillary carcinoma; MicroRNA-124; Cell proliferation; Cell migration; AKT serine / threonine kinase 1