Silence Atf5 Ovarian Cancer Cell Proliferation, Apoptosis And The Effect Of Chemotherapy Sensitivity

Objective To investigate the effect of ATF5gene silenced by dominant negative on growth, apoptosis and chemosensitivity of ovarian carcinoma cells.Method1. The expression of ATF5in different ovarian carcinoma cell lines were detected by RT-PCR and Western blot.2. The pLeGFP-C1-NTAzip-ATF5and pLeGFP-C1recombinant plasmids were respectively transfected into SKOV-3cells with LipofectamineTM2000reagent.3. MTT and Flow cytometry was respectively used to evaluate the proliferation and apoptosis rates of cells, which was interfered with the function of ATF5.4. The cells were exposed to cisplatin at different concentrations for24hours. The proliferation was assayed by MTT.5. After transfection24hours, the cells were treated with4umol/L cisplatin at different hours (Oh,12h,24h,48h), flow cytometry was used to detect apoptosis.6. The expression of bcl-2protein was detected by western blot.Results1. The expression of ATF5in SKOV-3was the highest among the three kinds of ovarian cancer cell lines, the expression difference was statistically significant (P<0.05).2. The recombinant plasmid can efficiently and specifically inhibit the expression of ATF5gene. Loss of the function of ATF5led to apoptosis of SKOV-3cells, especially in the early stage.3. Compared with the nonspecific and control group, the growth inhibition rates and the apoptosis rates of cells significantly increased in the specific transfection group when they exposed to cisplatin. Meanwhile, the expression of bcl-2was obviously down-regulated in group of transfected dominant-negative ATF5(P<0.05).Conclusion Interference with the function of ATF5could markedly inhibit cell proliferation, induce cell apoptosis and increase the sensitivity of SKOV-3to cisplatin. bcl-2was an ATF5-targeted apoptosis-related gene.