| ObjectiveTo investigate the effect of chemotherapy drugs on the expression of epithelial ovarian cancer tissue PI3K,P-AKT,PTEN for evaluating the clinical significance of neo-adjuvant chemotherapy in the treatment of advanced epithelial ovarian cancer to provide a theoretical basis.Methods1.Tissue collectionSpecimens from the First Affiliated Hospital of China Medical University gynecological surgery and pathology confirmed advanced ovarian serosity papillary cystadenocarcinoma.A total of 37 cases of(FIGO stages:StageⅢ35 cases,Ⅳtwo cases)from September 2006 to September 2007.All patients with cytoreductive surgery. 12 cases for the experimental group,Which will advance chemotherapy(1-3 cycles),as carboplatin chemotherapy(400mg/m~2,a single intraperitoneal injection)+ cyclophosphamide(600mg/m~2,intravenous)PC programme.No preoperative chemotherapy was 25 cases for the control group.Postoperative chemotherapy at PC programme,the same dosage and usage,including postoperative chemotherapy treatment for six to 30 cases,more than two courses of less than six to seven cases of treatment.As of March 2008,37 patients with one death.Based:the rapid admission of ovarian cancer 0.5 cm x0.5 cm two pieces respectively add an EP of 1ml RNA out reagents,and 1ml of lysis and were conserved in liquid nitrogen soon aider operation(within 5 minutes)for 4 hours then transported to -70℃deep freeze refrigerator to be conserved.2.MethodsThe expression of PTEN mRNA in epithelial ovarian carcinoma was evaluated by RT-PCR.Step includes:(1)Total RNA was extracted with RNA out Reagent,total RNA was prepared by solvent extraction following the instructions.(2)The reverse transcription reaction(RT)and polymerase chain reaction(PCR) were operated according to the direction of test kit.(3)The expression of mRNA was evaluated by semi-quantitative analysis.The expression of PI3K and P-AKT protein in epithelial ovarian was evaluated by western blot.Steps include:(1)The protein sample preparation.(2)SDS-polyacrylamide gel electrophoresis.(3)To the membrane.(4)Closed.(5)One anti-hybrid.(6)The anti-hybrid.(7)Chromogenic substrate.(8)Computer scanning technology analyzes the experimental result.3.Statistical AnalysisUnited States SPSS12.0 statistical analysis software system is used to analyze the result.The data were evaluated by independent-samples t test and Fisher exact probability.(α=0.05)Results1.Preoperative chemotherapy group of ovarian cancer PTEN mRNA expression level was significantly higher than that of no preoperative chemotherapy group,the difference between the two groups was significant(P＜0.01).2.Preoperative chemotherapy group of ovarian cancer PI3K protein expression levels were significantly lower than that of no preoperative chemotherapy group,the difference between the two groups was significant(P＜0.05).3.Preoperative chemotherapy group of ovarian cancer P-AKT protein expression levels were significantly lower than that of no preoperative chemotherapy group.The difference between the two groups was significant(P＜0.05).4.Preoperative chemotherapy satisfaction by surgery(residual disease＜2 cm) ratio was significantly higher than that of no preoperative chemotherapy group.The difference between the two groups was significant(P＜0.05).5.Preoperative chemotherapy group after two cycles of chemotherapy,CA125 significantly lower than the average level of no preoperative chemotherapy group,the difference between the two groups was significant(P＜0.01).6.In preoperative chemotherapy group,residual disease＜2 cm after six months in patients with clinical remission rate higher than the residual disease＞2 cm patients,the difference between the two groups was significant(P＜0.05).Conclusions1.Preoperative chemotherapy can be cut short course of treatment of ovarian cancer PI3K and P-AKT protein expression,while raising PTEN mRNA expression levels,and promote the anti-tumor cell apoptosis to the role.2.Neo-adjuvant chemotherapy can improve optimal operation rate,speeding up the decline in the level of CA125,optimal cyeoreductive surgery can improve patients recent remission rate. |
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