Effects Of Lentivirus Mediated AP-1 Gene Silencing On Proliferation,Apoptosis And Survivin Expression Of SKOV3 Cells

Objective:To investigate the effect of lentivirus mediated AP-1 gene silencing on the proliferation,apoptosis and survivin expression of ovarian cancer SKOV3 cells.Methods:1.Three groups of lentiviruses,AP-1-shRNA-1/2/3,were constructed by lentivirus vector mediated RNA interference technology,and their virus titers were measured.2.RT-PCR and Western blot were used to verify the AP-1 silencing effect of lentivirus in each group,and the best lentivirus was screened.3.SKOV3 cells infected with lentivirus with the best silencing effect were used as the experimental group,and SKOV3 cells infected with uninfected virus and infected with empty vector virus were used as the control group.The proliferation and apoptosis of SKOV3 cells were observed by MTT method and annexin v-APC method,and the expression of Survivin in SKOV3cells was detected by real-time fluorescent quantitative PCR and Western blot.Results:1.Three groups of AP-1-shRNA-1/2/3 lentiviruses have been successfully constructed by sequencing.The virus titer is more than 2×10~8TU/mL.2.The results of real-time fluorescent quantitative PCR and Western blot showed that AP-1 could be silenced at mRNA and protein levels,and AP-1-shRNA-3 had the best silencing effect for subsequent experiments.3.After AP-1 silencing,SKOV3 cells were inhibited in proliferation,increased in apoptosis,and decreased in Survivin mRNA transcription and protein expression.Conclusion:AP-1 gene silencing mediated by lentivirus can inhibit the proliferation and increase the apoptosis of SKOV3 cells.The mechanism may be related to the down-regulation of survivin.The specific mechanism needs further study.