Effect of microRNA-497 on the proliferation and apoptosis of baldder cancer cell line EJ / 实用医学杂志

ABSTRACT

Objective To investigate the effect of microRNA-497 (miR-497) on cell proliferation and apoptosis capability of baldder cancer cell line EJ. Methods EJ human baldder cancer cells were divided into miR-497 mimics group,mimics NC group,miR-497 inhibitor group and inhibitor NC group. MiR-497 mimics,mimics NC,miR-497 inhibitor and inhibitor NC were transfected into EJ cells by LipfectamineTM 2000. The transfection effi-ciency was observed under a fluorescence microscope 6 hours after. And qRT-PCR was used to detected the expres-sion of miR-49748 hours after. Plate clone formation assay ,MTT assay and flow-cytometric analysis of apoptosis were used to detect the cell proliferation and apoptosis of bladder cancer EJ cells. Western blot was employed to de-termine the protein expressions of bcl-2 and caspase-3. Results Under fluorescence microscope ,the efficiency rate of four groups were over 90%. And qRT-PCR results showed miR-497 expression in EJ cells increased signifi-cantly in miR-497 mimics group than those in the control group(P<0.01),while miR-497 inhibitor post expres-sion decreased(P<0.01). Overexpression of miR-497 significantly suppressed EJ cells proliferation(P<0.01), and prompted EJ cells apoptosis(P < 0.01)compared with the control group. Moreover ,opposite results were ob-tained when miR-497 inhibitor was transfected into EJ cells . Compared with negative control ,the protein expres-sion of Bcl-2 down-regulated(P < 0.01)and Caspase-3 up-regulated(P < 0.01)in miR-497 mimics transfection group. Conclusions MiR-497 could suppress the proliferation and promote apoptosis of EJ cells ,which might be related to the protein expression of Bcl-2 and Caspase-3.