| P53 gene is the most commonly mutated gene in ovarian tumors. There is P53 gene mutated among 30%～79% ovarian tumors. At present, wide-type P53 gene replacement therapy is among focus of tumor gene therapy, of which, recombinant adenovirus-P53 is the most commonly used. It has been reported that adenovirus-mediated wide-type P53 gene transfection into tumor cells can inhibit cell growth, promote cell apoptosis, and enhance chemotherapy sensitivity. This research is to evaluate the effects of domestic recombinant adenovirus-P53 (rAd-P53,Gendicine) on growth inhibition and apoptosis of human ovarian adenocarcinoma cell lines SKOV-3 and CAOV-3.Objective: The aim of the study is to evaluate the effects of domestic recombinant adenovirus-P53 (rAd-P53, Gendicine) on growth and chemosensitivity of human ovarian adenocarcinoma cell lines.Methods: Human ovarian adenocarcinoma cell lines CAOV-3 and SKOV-3 (mutant P53) were treated with rAd-P53, cisplatin (DDP) and rAd-P53 + DDP respectively. P53 expression was dectected by Western blot. The cell growth inhibition was assessed by MTT, and cell cycle and apoptosis were dectected by flow cytometry.1. use MTT to compare CAOV-3 and SKOV-3 cell growth inhibition rate when they were treated with rAd-P53, cisplatin (DDP) and rAd-P53 + DDP respectively of different time and dose. Compare tumor cell growth inhibition of different sequence of combination treatment.2. use flow cytometry to compare cell cycle and apoptosis of CAOV-3 and SKOV-3 when treated with rAd-P53 (100MOI), DDP (20uM) and rAd-P53 (100MOI) + DDP DDP (20uM) respectively after 72 hours.3. use Western blot to detect P53 expression of CAOV-3 and SKOV-3 after treated with rAd-P53 (100MOI) 72 hours later.Results: 1. There was a dose-dependent and time-dependent inhibition of cell proliferation by rAd-P53. After combined treatment with rAd-P53 (100MOI) and DDP (20uM) for 72 hours, the growth inhibition rate of CAOV-3 cells was 73.57±0.43%,which was significant higher than that in rAd-P53 group (45.53±0.95%, P<0.01) and DDP group (52.05±2.13%, P<0.01) . The growth inhibition rate of SKOV-3 cells was 74.70±0.86%, which was significant higher than that in rAd-P53 group ( 50.72±2.04%, P<0.01) and DDP group (59.46±1.17%, P<0.01) .The sequence of combined treatment is of no difference to the cellgrowth inhibition of SKOV-3 and CAOV-3.(P>0.6)2. Combined administration of rAd-P53 and DDP remarkably arrested CAOV-3 and SKOV-3 in G0/G1 (CAOV-3: 67.47±1.03 % , SKOV-3: 67.26±2.37%), which was significantly higher than that in control group (CAOV-3: 41.58±1.72%, SKOV-3 : 41.68±3.51 %, P<0.01) and DDP group (CAOV-3: 36.87±6.53%, SKOV-3: 48.36±9.34%, P<0.01).And cells in S phase significantly decreased (CAOV-3: 32.51±1.06%, SKOV-3: 32.53±2.32%), which was significantly lower than that in that in control group (CAOV-3: 54.65±2.79%, SKOV-3 : 54.94±0.65%, P<0.01), rAd-P53 group (CAOV-3: 36.10±3.67%, SKOV-3 : 37.11±1.20%, P<0.01) and DDP group (CAOV-3: 45.98±8.38%, SKOV-3 : 44.88±6.11%, P<0.01).Meanwhile the apototic rate of CAOV-3 cells was 19.39±1.67% in rAd-P53 + DDP group, which was significantly higher than that in rAd-P53 group (2.61±0.20%, P<0.01) and DDP group (6.61±0.49%, P<0.01). The apototic rate of SKOV-3 cells was 38.12±1.83% in rAd-P53 + DDP group, which was significantly higher than that in rAd-P53 group (5.70±0.47%, P<0.01) and DDP group (8.52±0.47%, P<0.01).3. High level P53 expression was dectected in CAOV-3 and SKOV-3 cells by Western blot after administeration of rAd-P53 (100MOI) 72 hours later.Conclusion: rAd-P53 can tranfect wide-type P53 into the genome group of human ovarian tumor cells, arrest tumor cells at G0/G1, inhibit the growth of human ovarian tumor cells, promote tumor cells apoptosis. Its combibation with DDP may significantly enhance the chemosensitivity of human ovarian tumor cells to DDP.
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