Clinical And Experimental Study Of The Effects Of Cyclooxygenase-2 And Its Related Factors On Ovarian Carcinoma

Objectives1. To detect the expression of COX-2, VEGF-C, and VEGF-A in all specimens including epithelial ovarian carcinoma, benign ovarian neoplasm and normal controls by immunohistochemical staining.2. To study the association of COX-2, VEGF-C, and VEGF-A with the clinicopathological charactors of epithelial ovarian carcinoma patient.3. To study the correlation of COX-2, VEGF-C, and VEGF-A, furthermore , To investingate their effects on MVD and MLD.4. To detect the expression of COX-2 in ovarian carcinoma cell lines by immunohistochemical staining. To investingate the effects of the selective COX inhibitor celecoxib on the proliferation and apoptosis of ovarian carcinoma cell lines by MTT and acridine orange and ethidium bromide staining methods.Methods1.The expression of COX-2, VEGF-C, VEGF-A, VEGFR-3, and CD34 were detected in 45 epithelial ovarian carcinoma, 10 benign ovarian neoplasm, and 12 normal ovarian tissues by immunohistochemical technique. The relationship of the expression between COX-2 and VEGF-C, VEGF-A were investgated. The relationship of their expression with the clinicpathologic characters was determined by statistics, the effectsof COX-2 and VEGF-C on MLD, COX-2 and VEGF-A on MVD was measured2. The expression of COX-2 was detected in ovarian carcinoma cell lines SKOV3 and OVCAR3 by immunohistochemical staining, the effects of the selective COX inhibitor celecoxib on the proliferation and apoptosis of ovarian carcinoma cell lines was determined by MTT and acridine orange and ethidium bromide staining methods. Results1. 32 cases exhibited positive expression of COX-2 in 45 epithelial ovarian carcinoma tissues, 13 cases were lower expression and 19 cases were higher expression. The positive rate was 71.1%. The expression of COX-2 was not found in benign ovarian neoplasm tissues and normal controls. The positive expression rate of VEGF-C was 51.1 % in EOC tissues. There were 11 cases exhibited lower expression and 12 cases were higher expression. The expression of VEGF-C in ovarian malignant was higher than that in benign ovarian neoplasm tissues and normal controls(x2= 4.37, PO.05-, x 2=4.08,P<0.05). The positive rate of VEGF-A was 55.6% in EOC tissues. There were 9 cases exhibited lower expression and 16 cases were higher expression, The expression of VEGF-C in ovarian cancer was higher than that in benign ovarian neoplasm tissues and normal controls( x 2 = 4.29,P<0.05; x 2 = 4.81,P<0.05),2 cases in benign ovarian neoplasm tissues and 2 cases normal ovarian tissues were positive expression only.2. The positive expression of COX-2, VEGF-A and VEGF-C had no relationship with age, clinical stage ,pathologic defferentiation ,histological types, and hydroperitoneum ( respectively P>0.05 ) . Their expression respectively had significantly relationship with lymphnode metastasis (respectively PO.05) . The positive expression rate of COX-2, VEGF-C, and VEGF-A in cases with lymphnode metastasis (respectively 83.3%, 66.7%, 66.7%) was respectively higher than that with non-lymphnode metastasis (respectively46.7%, 20%, 33.3%).The expression of each marker was statistic diference between the two groups (respectively PO.05) . MLD was 18.5±7.8 in cases with lymphnode metastasis and 12.3±5.7 in non- lymphnode metastasis. There were statistic diference between the two groups ( P<0.05, t=2.73) . MVD was 46.1±20.5 in cases with lymphnode metastasis and 32.4±17.6 in non-lymphnode metastasis. There were statistic diference between the two groups also(PO.05, t=2.21) .3. Spearman rank correlation analysis showed that there were a positive correlation between expression of COX-2 and VEGF-C (rs=0.415, PO.01), COX-2 and VEGF-A (rs=0.624, P<0.01) .The mean MLD was 18.7±7.3 in COX-2 positive group,and it was 9.5±7.1 in COX-2 negtive group, there were sigificant diferences(t=3.86,P<0.01) . The mean MVD was 42.5±15.4 in COX-2 positive group, and it was 28.4±17.5 in COX-2 negtive group, there were sigificant diferences (t=2.68,P<0.01) .4. The growth of 0VCAR3 and SKOV3 cell lines were inhibited by celecoxib in a dose- and time- dependent manners. The treated 0VCAR3 cells exhibited some mophologycal features of apoptosis. The apoptotic index of the treated 0VCAR3 was in a dose- and time- dependent manners,S-N-K analysis displayed statistic diference between every two groups of them( respectively PO.05).Conclusions:1. The expression COX-2 was found in EOC tissues and its cell lines, and was not found in benign ovarian neoplasm tissues and normal controls.2. COX-2 may stimulated angiogenesis through VEGF-A up-regulation in ovrian epithelial carcinoma.3. COX-2 may stimulated lymphangiogenesis in EOC. VEGF-C maybe its critical media factor.4. lymphangiogenesis and angiogenesis induced by COX-2 may have a important role in the progression of lymphnode metastasis of malignant tumors.5. The growth of 0VCAR3 and SKOV3 cell lines was inhibited by celecoxib in a dose- and time- dependent manners, the apoptotic index of 0VCAR3 was in a dose-and time- dependent manners also. Celecoxib may be used as chemotherapeutic agents for ovarian cancer.