MicroRNA-183 Promote The Drug Resistant, Invasive And Migration Of Synovial Sarcoma Via Downregulation Of The Expression Of EGR1 In Human Synovial Sarcoma Cells Line SW982

Objective:To investigate the influence of drug resistant, invasive and migration of synovial sarcoma via regulating the expression of microRNA-183 in Human Synovial Sarcoma Cells line SW982 and analyze the possible potential target:EGR1. Methods:We changed the expression of miR-183 by transfecting the miR-183 mimics, inhibitor and N.C to SW982 cells line. Transfection efficiency was detected by using RT-PCR and FACS. MTT was used to detect the cell proliferation after transfection. We cultured the SW982 cells withlO μmol/l doxorubicin and measured the inhibition rates after 24h,48h and 72h by MTT. FMC was used to the detection of cell cycle of SW982 cells after transfection. Using Annexin V-FITC/PI to measure apoptosis of cells after transfection. Transwell model was used to exam the change of migratory and invasive abilities after transfection. Western blot was used to analyze the expression of EGR1 after transfection. Using a dual-luciferase report assay to identify whether EGR1 is the direct target of miR-183. Results:Real-time PCR analysis revealed that the expression of miR-183 was higher in mimics group (75.30±9.90) compared to the normal groups (1.157±0.52)(P<0.001). The miR-183 expression in inhibitor (0.22±0.09) was lower than normal groups (P<0.001).In normal and negative control (N.C) groups, the result was similar(.P=0.637). In the research of proliferation, apoptosis and cell cycle of SW982 cells after transfection, the result shows that there was no significant difference. In the study of drug resistant, after transfection and 24 hours’exposed in doxorubicin, the suppression ratio of mimics groups (0.0051±0.022) was much lower than the normal groups (0.0903±0.039)(P=0.003), and he suppression ratio of inhibitor groups (0.1351±0.062) was much higher than the normal groups (0.0281±0.033) (P=0.009). Transwell analysis revealed that, in migratory study, the number of SW982 cells penetrated this membrane in mimics and inhibitors group was 108.61±4.94,54.36±4.03, respectively(P<0.001). In invasive study, the number of SW982 cells penetrated this membrane in mimics and inhibitors group was 79.13±5.77,20.87±3.17, respectively (P=0.036). The expression of EGR1 was detected by western blotting, the result shows that in mimics group (286.59±32.77) the relative expression of EGRl was dramatically decreased compared with inhibitor group (373.00±38.40)(P<0.001). In inhibitor group (496.93±42.61) the expression of EGRl was dramatically increased compared with mimics group (P=0.012). We further identified EGR1 as a direct target of miR-183 by the dual-luciferase report assay. Conclusion:miR-183 regulated the drug resistant, invasive and migration of synovial sarcoma and there were a potential target:EGR1. miR-183 had no influence on the proliferation, apoptosis and cell cycle of SW982 cells.