RNA interference gene therapy targeting NHERF1 inhibits proliferation of prostate cancer cell line PC-3M

Abstract

Objective To investigate the effect of NHERF1-shRNA on malignant behaviors of prostate cancer cell PC-3M. Methods pSuper.puro NHERF1 shRNA vector and negative control (pSuper.puro luciferase shRNA) were transfected into prostate cancer cell PC-3M. Then the cell line stably knockdown NHERF1 was obtained and verified by Western blot analysis. Methyl thiazolyl thiazolium (MTT) was used to detect the proliferative activity of PC-3M cell. The cell migration abilities were examined by wound healing assay. Flow cytometry method was applied to detect the effect of NHERF1-shRNA on apoptosis of PC-3M cell line. The expressions of Bax and Bcl-2 in NHERF1-shRNA cells were detected by Western blot. Results Compared with PC-3M cells and the cells which were integrated of empty vectors, NHERF1-shRNA could weaken the proliferation of PC-3M cells significantly (F=62.63, P=0.004), and significantly repress PC-3M cells migration, and increase the apoptosis percent of PC-3M cell line apparently [(2.55±0.92)% and (11.98±3.28)%], up to 4 times (t=-2.392, P=0.001). NHERF1-shRNA could reduce the expression of Bcl-2 and increase the expression of Bax. Conclusion NHERF1-shRNA can inhibit the proliferation and migration of PC-3M cell, and promote its apoptosis. Key words: Prostatic neoplasms; Apoptosis; NHERF1