The Inhibitation Expression of HDAC2 Gene Intereferred by Lentivirus Mediated RNA

Objective: To constuct the HDAC2 gene lentivirus vector and to verify the expression of HDAC2 gene silencing in hepatocellular carcinoma Hep G2 cells. Methods: Correctly identified three pairs of recombinant plasmid p LKO. 1-HDAC2-shRNA1,2,3,one pair of negative control p LKO. 1-HDAC2-shRNAneg and empty plasmid p LKO. 1 were co-transfected respectively with packed plasmids ps PAX2 and envelope plasmid p MD2. G into human embryonic kidney cells 293 T to produce infectious lentiviral particles by liposome method. The supernatant containing virus particles were collected,purified and infected with Hep G2 cells. Untreated Hep G2 cells were blank control group. HDAC2 gene mRNA and protein expression levels in Hep G2 cells were detected by Real-time PCR and Western blotting respectively 48 h and 72 h after infection. The interfering effect of recombinant plasmids in each group was indentified and analyzed. Results: Real-time PCR results showed that compared with control group,3 interference sequences transfected into Hep G2 liver cancer cells could significantly inhibit HDAC2 mRNA expression( P 0. 05),of which p LKO. 1-HDAC2-shRNA1 showed more obvious inhibition rate,reaching 82. 09%. Western blotting test results showed that 3 interference sequences could significantly inhibit HDAC2 protein expression( P 0. 05),of which p LKO. 1-HDAC2-shRNA1 and p LKO. 1-HDAC2-shRNA3 showed more obvious interference effect,and the difference between them was not statistically significant( P 0. 05). Conclusion: The constructed recombinant plasmid p LKO. 1-HDAC2-shRNA can effectively restrain HDAC2 gene expression in Hep G2 cells in the transcription level and protein level,which provides a useful tool for follow up study on HDAC2 gene function.