Part1:Expression Of POSTN Promoting Metastasis And Regulated By Malignant Ascites In Ovarian Cancer Part2:Prognostic Analysis Of Bulked Locally Advanced Cervical Cancer Without Pathological High-risk Factors Treated With Neo-Adjuvant Chemotherapy And Radi

Objective A number of ovarian cancer patients were primarily diagnosed with malignant ascites,and most were in late stage with intraperitoneal metastasis.To predict the status of metastasis using malignant ascites and develop a treatment target was the main foucs in clinical procedure.POSTN has recently been reported to overexpress in many malignant carcinomas and be related to high invasiveness.However,its expression status and role in ovarian carcinoma was uncertain.This study aimed to analyse POSTN expression and its role in ovarian carcinoma.Methods This study included six parts.1.Measurement of POSTN expression in ovarian cancer.We first searched the GEO database and analysed the expression of POSTN gene in ovarian carcinoma.Tissue array was made from surgical specimens of 167 ovarian serous adenocarcinoma patients primarily treated in our hospital and immunohistochemistry was performed to evaluate the expression of POSTN protein.The serum concentration of POSTN protein in both ovarian cancer patients and normal people was measured by ELISA and compared by t-test.2.Analysis of the predict potential of POSTN in the prognosis of ovarian cancer.Establish the clinical database after follow-up.Correlation analysis was conducted between the expression of POSTN and the common clinical and pathological factors in ovarian cancer.The overall survival and progression free survival were analysed with Kaplan-Meier method.Log-rank test and Cox regression model were used to evaluate whether POSTN could act as a prognostic risk factor.3.Analysis of the relation between POSTN and common molecules of EMT.Pearson correlatin test was performed between the expression of POSTN and common factors of EMT in TCGA ovarian carcinoma database.QPCR was performed to measure level of MMP2 and Cadherinl 1 after knockdown of POSTN in ovarian cancer cells.4.Functional experiment of POSTN to promote metastasis in vitro.TOV-112D cells were disposed with human recombinant POSTN protein and POSTN antibody respectively,Transwell migration assay was performed to evaluate the migration function regulated by POSTN.After knockdown of POSTN with siRNA in TOV-112D,Transwell migration assay was performed to evaluate its migration potential.5.Functional experiment and mechanisms of malignant ascites to promote metastasis.After disposal of malignant ascites in SKOV3 cell line,Transwell migration assay was performed to evaluate its migration protential.To discover the regulation of POSTN by ascites,Western Blotting was performed to evaluate the expression of POSTN protein.Furthermore,ELISA was performed to detect the POSTN protein in the ascites to know whether it could be secreted into the ascites to promote intraperitoneal metastasis.6.The regulation correlation between POSTN and ?-catenin.POSTN plasmid was transfected into SKOV3 cell line,and the expression of POSTN and ?-catenin protein was measured by WB.Their expression was also measured after the treatment of malignant ascites and POSTN antibody.Results 1.Data based on GSE6008 revealed the overexpression of POSTN in ovarian carcinoma compared to normal ovary with significant difference(P=0.026).IHC of tissue array revealed that POSTN mainly expressed in the stroma and epithelial cells,especially in the stroma.Compared to the normal people,the serum concentration of POSTN protein was significantly higher in the ovarian cancer patients(P<0.0001).The expression of POSTN in ovarian metastatic carcinoma was significantly higher than primary carcinoma in both mRNA level(P=0.023)and protein level(P=0.03).2.Survival analysis based on GSE14764 database showed that OS and PFS were significantly shorter in the patients with higher POSTN expression.Survival analysis based on our tissue array and clinical database revealed that the 5 year OS and PFS rate of higher POSTN group was 34.4%and 14.1%respectively,both of which were significantly lower than lower POSTN group(P=0.016,P=0.039).However,Cox regression analysis only showed the FIGO stage(P=0.006)and CA125 level on initial treatment(P=0.044)as independent prognostic factors.3.Data based on TCGA ovarian carcinoma database showed that the expression of POSTN was significantly related to common epithelial-mesenchymal transition factors.And qPCR analysis revealed the concurrent downregulation of MMP2 and Cadherin11 after knockdown of POSTN.4.In vitro,the migration potential of TOV-112D cell line improved after disposal of human recombinant POSTN protein(P=0.02)and reduced after treatment of POSTN antibody(P=0.02)and knockdown of POSTN by siRNA(P=0.002).5.After treatment of malignant ascites,upregulation of POSTN was detected and migration potential enhanced in SKOV3 cell line.High concentration with average 214.9ug/ml of POSTN protein was detected in the ascites,which means the POSTN protein could be secreted into the ascites to promote the intraperitoneal metastasis.6.The expression of?-catenin was upregulated after the overexpression of POSTN in SKOV3 cell line and downregulated after the knockdown of POSTON in TOV-112D cell line.The expression of ?-catenin could be blocked with POSTN antibody.The stimulating effect of malignant ascites in tumor migration was counteracted via ?-catenin.Conclusions In conclusion,POSTN was highly expressed in ovarian carcinoma,and its overexpression carcinoma in stroma could predicted poor prognosis.The expression of POSTN in ovarian metastatic carcinoma was higher than primary carcinoma.Overexpression of POSTN and rhPOSTN protein could improve the migration potential of ovarian cancer cells.The malignant ascites could promote the migration potential of ovarian cancer,and the overexpression of POSTN protein could be secreted into the ascites to accelerate the intraperitoneal metastasis.?-catenin was positively regulated by POSTN in the metastasis of ovarian cancer.Objective To analyze the prognosis of bulked locally advanced stage cervical squamous cancer(Ib2 and IIa2 stage)without high risk factors after treatment of neo-adjuvant chemotherapy(NACT)and radical hysterectomy.Methods This retrospective study included patients with International Federation of Gynecology and Obstetrics(FIGO)stage Ib2 and IIa2 cervical squamous cancer without high risk factors who underwent platinum-based NACT followed by radical surgery from January 2008 to January 2015.The responses of NACT were observed and compared in their effect on postoperative pathologic risk factors.Kaplan-Meier method and Cox regression analysis were performed to analyze survival status,Results 282 patients with the average age of 44.4±6.7 were recruited into this study.After NACT,42 patients achieved complete response(CR,14.9%),while 138 patients achieved partial response(PR,48.9%)and 102 stable disease(SD,36.2%).The rate of pathologic diameter>4 cm,deep stromal invasion(DSI)positive and lymph-vascular space invasion(LVSI)positive rate decreased significantly in CR and PR group compared with SD group(P<0.05).The rate of multiple intermediate risk factors was 0%vs 13.8%vs 45.1%respectively(P=0.000).The disease relapsed in 23 patients,and 12 died.On the whole series,5-year overall survival rate was 91.7%,and 5-year recurrence-free survival rate was 88.9%.The rate of recurrence(P=0.002)and mortality(P=0.036)were higher in LVSI positive patients compared with LVSI negative.And the rate of recurrence was higher in patients with multiple intermediate risk factors,compared with no or one intermediate risk factors(P=0.002).Univariate analysis revealed that LVSI positive and multiple intermediate risk factors were the factors predicted recurrence and mortality(P<0.05),and no significance in age,stage,tumor grade,tumor diameter before or after NACT,response to NACT,or DSI positive factors(P>0.05).On multivariate analysis,the factor of primary tumor diameter?6cm(P=0.022)and multiple intermediate risk factors(P=0.001)were independent prognostic variables for recurrent-free survival.Besides,multiple intermediate risk factors was independent prognostic variable for overall survival(P=0.034).After surgery,175 among 282 patients received adjuvant therapy with 26 radiotherapy,46 chemotherapy and 103 concurrent radiochemotherapy.On survival analysis of postoperative adjuvant treatment,5-year recurrence-free survival rate of radiotherapy group was significantly lower in patients with the factor of SD response to NACT(P=0.011)and multiple intermediate risk factors(P=0.008),while overall no significance in overall survival rate.Conclusions NACT played beneficial role in relieving the status of intermediate risk factors in stage Ib2 and IIa2 cervical squamous cancer without high risk factors.The status of multiple intermediate risk factors was independent prognostic factors for recurrence and mortality.For patients with multiple intermediate risk factors,postoperative adjuvant chemotherapy or concurrent radiochemotherapy might be the better choice to prevent relapse.