Effects of microRNA-149-5p modulating serine-arginine protein kinase 1 on invasion and migration of renal cancer cells

Objective To elucidate the role and mechanism of tumor suppressor microRN (miRNA, miR)-149-5p in metastasis potentials of renal cell carcinoma. Methods GRC-1 cells were divided into miR-149-5p group(transfected with miR-149-5p mimics), miR-NC group(transfected with mimics control), miR-149-5p+ SRPK1 group(co transfected with miR-149-5p mimics and pcDNA3.1-SRPK1), miR-149-5p+ vector group(co transfected with miR-149-5p mimics and pcDNA3.1). The transfection efficiency of miR-149-5p was detected by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR). The cell invasion and migration ability were measured by Transwell chamber. The levels of N-cadherin and E-cadherin proteins were detected by Western blotting. SRPK1 may be a target gene of miR-149-5p by using online target gene prediction software. Targeting relationship was identified by double luciferase reporting system. Results The levels of miR-149-5p (2.69±0.27 vs. 1.01±0.08) and E-cadherin (0.81±0.09 vs. 0.42±0.05)in the miR-149-5p group were significantly higher than those in the miR-NC group, and invasion number [(70.81±6.35) vs. (110.64±9.67)], migration number [(108.46±9.27)vs. (162.76±12.71)], N-cadherin (0.46±0.05 vs. 0.79±0.11)and SRPK1 (0.45±0.06 vs. 0.92±0.08)proteins levels were significantly lower than miR-NC group, and the difference was statistically significant (FmiR-149-5p=95.385, Finvasion number=20.216, Fmigration number=22.010, FN-cadherin=15.100, FE-cadherin=31.331, FSRPK1=31.785, P 0.05). The level of E-cadherin (0.45±0.04 vs. 0.80±0.08) in the miR-149-5p+ SRPK1 group was significantly lower than that in the miR-149-5p+ vector group, and the invasive number [(99.51±7.48) vs. (71.84±5.37)], migration number [(145.06±11.14) vs. (107.63±10.20)], N-cadherin (0.86±0.10 vs. 0.47±0.04) and SRPK1 (0.89±0.06 vs. 0.50±0.07) proteins levels were significantly higher than the miR-149-5p+ vector group, and the difference was statistically significant (tSRPK1=7.327, tinvasion number=5.205, tmigration number=4.292, tN-cadherin=6.272, tE-cadherin=6.778, P<0.05). Conclusion Tumor suppressor miR-149-5p targeting negative regulation of SRPK1 expression reduces the migration and invasion of renal cancer cells. Key words: MicroRNA-149-5p; Serine-arginine protein kinase 1; Renal cancer; Metastasis; Invasion