Influence On Cell Apoptosis By SiRNA Induced Silencing Of HDAC1Gene In Bladder Cancer RT112Cells

Objective: To observe the effect of silencing of histone deacetylases1(HDAC1) by siRNA on the apoptosis in bladder cancer RT112cells，andits possible mechanism is discussed preliminarily.Methods: Culture human bladder cancer RT112cell in vitro, theexperiment has4group：normal group, the HDAC1-control group, theHDAC1-siRNA group and SAHA group. Before building HDAC1-siRNArecombinant plasmid，design2HDAC1-siNRA sequences and1controlsequence.Select the HDAC1-siRNA-2sequence which has highersilencing efficiency after respectively transfecting RT112cell to do thefuctional experiment.Using MTT to detect the inhibition of SAHA on cellproliferation. Detect the apoptosis rate of four groups by Flow cytometry.Respectively extractinng mRNA and protein, detect HDAC1, Bax, Bcl-2gene mRNA expression with real-time PCR, and detect the expression ofHDAC1, PTEN, Bax, Bcl-2protein with Western blot.Results:(1) Compare with untreated group, the mRNA and proteinexpression of HDAC1and Bcl-2in RT112cell is significantly reduced inHDAC1-siRNA and SAHA groups. There was no significant difference among groups on the expression of Bax. But the expression ofPTEN protein was improved in HDAC1-siRNA and SAHA groups.（2）Flow cytometry analysis of apoptosis is increased.Conclusion:1.The HDAC1-siRNA recombinant plasmid that wasbuilded can disturb the mRNA and protein expression of HDAC1in vitro.2. HDAC1siRNA recombinant plasmid can induce the apoptosis ofRT112cells, and it may be related to promoting the level of histoneacetylation, tumor suppressor gene PTEN expression and reducing theratio of Bax/Bcl-2.