Effect of microRNA-9-5p targeting MEF2C on biological behaviors of alveolar rhabdomyosarcoma cells / 吉林大学学报(医学版)

ABSTRACT

Objective: To investigate the effect of microRNA-9-5p (miR-9-5p) targeting the myocyte enhancer factor 2C (MKF2C) on the biologicals behaviors of the alveolar rhabdomyosarcoma (ARMS) cells, and to provide the basis for the molecular diagnosis and targeted therapy of ARMS. Methods: The expression levels of miR-9-5p and MKF2C mRNA in ARMS tissue and cells were detected by qRT-PCR method' the proliferation rate of cells was detected by CCK-8 method, the apoptotic rate was detected by flow cytometry, the numbers of invasion and migration cells were detected by Transwell chamber assay, the luciferase activity in 293T cells was detected by double luciferase reporter gene, and the expression level of MEF2C protein in the cells was detected by Western blotting method. Results: The expression levels of miR-9-5p in ARMS tissue and cells were higher than those in normal skeletal muscle tissue and HSKMC cells ( Pcells in miR-9-5p inhibitor group was decreased ( Pcells in miR-9-5p inhibitor group was decreased ( Pcells were decreased ( P<0. 05). The luciferase activity in the 293T eels after co-transfection with MKF2C-3-UTR-WT and added with miR-9-5p was decreased (PmRNA and protein in the RH30 cells in miR-9-5p inhibitor group were increased (P'<0. 05). The expression level of MKF2C mRNA in the ARMS tissue was lower than that in normal skeletal muscle tissue ( PARMS tissue ( r= 0. 5420, P<0. 05); the expression level of MKF2C mRNA in the RH30 cells and PLA802 cells was lower than that in the HSKMC cells (P'-C0.01). Compared with transfection with control plasmid (KV), the expression level of MKF2C mRNA in the RH30 cells after transfection with MEF2C over-expression plasmid was increased ( P<"0. 01), the proliferation rate was decreased (P<.0.01), the apoptotic rate was increased (P<0.05), the number of invasion cells was decreased ( P<0. 05), and the number of migration cells was decreased ( Ptransfection with si-NC, the expression level of MEF2C mRNA in the RH30 cells after transfection with MKF2C siRNA was decreased ( Pcells was increased ( P<0. 01), and the number of migration cells was increased (P<0. 01).

Conclusion:

MiR-9-5p can directly target the 3-UTR of MEF2C mRNA to induce mRNA degradation and inhibit the expression of MKF2C, and promote the proliferation, invasion, migration and anti-apoptosis ability of the RH30 cells.