Regulation Of RB1and E2F1Gene Expression By ShRNA Induced Silencing Of HDAC1Gene In Prostate Cancer PC-3M Cells

Prostate cancer is one of the most common male malignancies in Eu-rope and America. Although the incidence of prostate cancer in China andother Asian countries is much lower than that in Europe and America, theincidence of prostate cancer in men is growing in recent years with thechange of life and eating habits.As a new class of anticancer drugs, histone deacetylase inhibitors(HDACIs) have the features of low toxicity and high efficiency, inhibitingtumor occurrence and development by inhibiting the proliferation of tu-mor cells and cell cycle, and accelerating apoptosis, etc. Studies haveshown that over-expression of HDAC1was detected in prostate cancertissues and cells. E2F1can regulate gene transcription by recruiting RB1and HDAC1and forming complexes. But HDAC1’s exact regulatory rolein this complex is uncertain.In this study, we constructed recombinant plasmid shRNA HDAC1to interference HDAC1gene expression. Recombinant plasmid shRNAHDAC1were successfully imported in PC-3M cells. After transfection,we detected the mRNA expression levels of RB1and E2F1gene byReal-time PCR and the cell cycle with Flow Cytometry.The results showed that after reducing HDAC1gene expression, RB1and E2F1genes mRNA expression levels were declined in varyingdegrees with no significance. The results suggest that HDAC1is not atthe regulation center in the complex. RB1, E2F1and HDAC1play a rolein regulation of cell cycle within the cell by forming complexes. RB1protein is involved in cell cycle regulation through the change of phos-phorylation and dephosphorylation form. Further protein level researchwill continue. After reducing HDAC1gene expression, cell cycle was in-hibited at G1phase, thereby apoptosis of tumor cells was promoted. Theresults suggest that HDAC1can be an effective molecule target of tumorinhibitor.In this study, we obtained the following conclusions:1. We successfully constructed two recombinant plasmids shRNAHDAC1to interference HDAC1gene expression using RNAi technology,which provided the basic material for the study on prostate cancer genetherapy.2. After reducing HDAC1gene expression, RB1and E2F1genesmRNA expression levels were declined in varying degrees with no signi-ficance.3. After reducing HDAC1gene expression, cell cycle was inhibitedat G1phase, thereby apoptosis of tumor cells was promoted.