Study Of Transduction Of AdA-fos On Reversal Of Cisplatin Resistance In Ovarian Carcinoma Cell Line 3AO/DDP

Objective: We aimed to observe the reversal effects and further exploit mechanisms of AdA-FOS on drug resistance in ovarian carcinoma cell 3AO/DDP in vitro. And we tried to reveal a new target and experimental evidence for reversal of drug resistance in ovarian carcinoma.Methods: First AdA-fos plasmids were used to transform the competence E. coli DH5α. Positive clones were confirmed by PCR evaluation. DNA ladder results proved that AdA-fos expressed correctly in DH5 α . We named the plasmids PCDNA3. 1/ AdA—fos. After purified and quantitated for transgenic using they were transferred into cultured 3AO/DDP cells by FuGENE6 transfection reagents . The new-built cells 3AO/DDP cells and other 3AO/DDP cells which were transfected by PCDNA3. 1 only grouped as experiment group and control groups . RT-PCR and DNA Agarose Gel electrophoresis was used to detect the expression of gene. Then cells were cultured in DDP separately. Cell morphologic changes were observed under the inverted phase contrast microscope . DNA Agarose Gel electrophoresis wasemployed for the cell apoptosis. Detected the apoptosis of 3A0/DDP cells after transfection by inmunohistochemystrial straining with anti-ssDNA mAb. And evaluated the MDR reversal of AdA-fos according to the apoptosis rate.Results: With moleucular cloning technuque, eukaryotic expression vectors of PCDNA3. 1/ AdA-fos were transducted into 3AO/DDP cells by using FuGENE6 mediation. RT-PCR and DNA Agarose Gel electrophoresis showed the expression of AdA-fos gene in 3AO/DDP cells. After AdA-fos treatment under the effect of DDP the inhibitory of proliferation was obvious and typical morphological characteristics of apoptosis including apoptosis bodies and the trapeziform bind of electrophoresis were able to be observed. Theapoptosis rate is 45.71 % which is higher than that ofcontrol groups . (P<0.005)Conclusion: AdA-fos could reverse drug resistance of ovarian carcinoma cells 3A0/DDP by inhibiting the combination of AP-1 and DNA so as to increase chemotherapeutic sensitivity of ovarian carcinoma MDR cells. The transduction of AdA-fos inhibited the growth of 3A0/DDP cells and induced apoptosis. AP-1 was an important target for ovarian carcinoma to reverse DDP resistance.