Knockdown Of PLCε Inhibits Cell Proliferation Via PTEN/AKT Signaling Pathway In Human Prostatecancer

PART ONETHE PREPARATION OF THE LV-SHPLCE PROSTATE CANCER CELL LINEObjective and methods:The lentivirus-shPLCε(LV-shPLCε)was designed to silence PLCε expression in DU 145 and PC3 cell lines,and the effectiveness was tested by qRT-PCR and Western blotting.MTT assay and colony formation assay were conducted to observed cell proliferation.Results:The lentivirus-shPLCε effectively knocked down the expression of PLCε in both gene and protein levels,MTT and colony formation assay were conducted to observed after Silencing the PLCs gene significantly inhibited cell proliferation in DU 145 and PC3 cell lines.Conclusion:Silencing the PLCε gene significantly inhibited cell proliferation in DU 145 and PC3 cell lines.PART TWOTHE CORRELATION BETWEEN PLCE AND PTEN IN PROSTATE CANCERObjective and methods:A total of 15 benign prostatic hyperplasia(BPH)tissue samples and 40 prostate cancer(PCa)tissue samples were obtained from patients,immunohistochemical staining were performed to test the expression levels of PLCε and PTEN,statistic methods were used to test the correlation between PLCε and PTEN.Results:Our results showed a higher expression of PLCε in approximately 90%of the PCa tissue samples compared to BPH tissues.PTEN had been identified as a tumor suppressor in prostate cancer and we also observed that the expression of PTEN was strongly upregulated in approximately 73.3%BPH tissues,but PTEN showed a low or undetectable level in PCa tissue samples,the correlation between the PLCε increase and the PTEN decrease in PCa tissue appeared a strong level of agreement.Conclusion:there was a great level of agreement between PLCε and PTEN in human prostate cancer tissues.PART THREE PLCE PROMOTES CELL PROLIFERATION VIA PTEN/AKT SIGNALING PATHWAY IN HUMAN PROSTATE CANCERObjective and methods:qRT-PCR and Western blotting were conducted to test the expression of PLCε,PTEN,AKT and P-AKT in both DU145 and PC3 cells,besides,Western blotting and immunofluorescence assays were used to detect changed PTEN expression in DU 145.Results:After being infected by LV-shPLCε,we noticed that PTEN expression was upregulated in DU 145 cells but not in PC3 cells.Furthermore,we found that PLCε gene knockdown decreased P-AKT protein levels,but AKT protein levels were not affected.Immunofluorescence assays showed that PTEN expression had intracellular distribution change in the DU 145 cell line,and Western blot analysis showed that PTEN was obviously upregulated in the cell nucleus and cytoplasm.Conclusion:PLCε is an oncogene,and knockdown of expression of PLCε inhibits PCa cells proliferation via the PTEN/AKT signaling pathway.