The Effects Of Silencing PICK1on Proliferation And Apoptosis Of Human Breast Cancer Cell Line MCF-7via RNA Interference

Objective:To explore the effects of silencing PICK1on proliferation and apoptosis of human breastcancer cell line MCF-7via RNA interference, and investigate whether the PICK1gene can be used as a potential target for breast cancer therapy.Methods:Design and synthesis siRNA for PICK1gene. Experiment was divided into three groups:negative control group, blank plasmid group and PICKl-siRNA group). siRNA was tansfected into MCF-7cells by Lipofectamine2000.(1)Transfection efficiency using fluorescence microscopy.(2)MTT assays were used to detect cell proliferation.(3) Western blot detecte expression of transfected PICK1, Bax and caspase-3protein.(4)Invasion using transwell assay.(5)Flow cytometer was used to observe apoptosis rate.Results:(1)Under the fluorescence microscope, the green fluorescence with the cells could be seen after transfection, suggesting the transfection was successful. The transfection efficiency was greater than70percent.(2)After48hours of transfection, The expression of PICK1protein in PICKl-siRNA group(0.11±0.04) was significantly lower than that in the negative control group(0.34±0.03) and blank group(0.36±0.02)(P<0.05).(3)MTT showed that the cell proliferation was reduced significantly in PICKl-siRNA group, compared with the negative control and blank in48h and72h after transfection(P<0.05). (4)Transwell experiment showed that the number of cells was reduced significantly in PICKl-siRNA group compared with the negative control and blank after transfection(P<0.05).(5)Flow cytometer showed that the apoptosis rate of MCF-7cells was increased significantly in PICKl-siRNA group compared with the negative control and blank after transfection(P<0.05).(6)Western-blot analysis indicated that the expression of Bax and caspase-3protein were up-regulated after transfection(P<0.05).Conclusion:siRNA targeting PICK1can inhibit MCF-7cell proliferation, decrease the cell invasive and induce apoptosis. its anti-tumor activity may be applicable on the treatment of human breast cancer.