The Study On Relationship Between Matrix Metalloproteinases And Tissue Inhibitors And Invasion And Metastasis In Ovarian Carcinoma

Ovarian cancer is one of the most common malignant tumor in women. The majority ofovarian cancers are diagnosed in their advanced stage and the relative 5-year survival rate is low.Invasion and metastasis is one of most important character of ovarian cancer. Matrixmetalloproteinases(MMP) are known to play an important role in cancer cell invasion bymediating the degradation of extracellular matrix (ECM).The activity of MMPs are regulated bytissue inhibitor of metalloproteinases(TIMPs) . The imbalance between MMPs and TIMPs hassignificant relevance with the invasion and metastasis of ovarian cancer. In this study, we soughtto explore the relationship between MMPs and TIMPs expression and clinic-pathologic patternin ovarian cancer and also to explore the role of MMP-9 in invasion and metastatic of ovariancancer.In this study, the reverse transcription polymerase chain reaction (RT-PCR)was used todetect the mRNA expression of MMP-9,2,7 and TIMP-1,2,3 in 48 cases of ovarian malignanttumor ,21 cases of benign ovarian tumor and 22 cases of normal ovarian tissues . Therelationship between MMPs and TIMPs expression and clinic-pathologic pattern and prognosisin ovarian cancer was analysed by Kaplan-Meier estimates and Cox regression model. Theresults showed the positive expression rate of MMP-9 and the half-quantity value ofMMP-9,MMP-2 were significantly higher in malignant tumor than that in normal control(P<0.05);The positive rate and half-quantity of TIMP-2,MMP-7,TIMP-3 mRNA weresignificantly higher in ovarian malignant and benign tumor tissues than those in normal control ;The ratio of MMP-9/TIMP-1 was higher in ovarian malignancies tissues(0.91±0.67) than thatin normal ovarian tissues (0.14±0.33) (P<0.05); The expression of MMP-9 mRNA washigher in patients with stage Ⅲ-Ⅳovarian cancer (1.13±0.66)than those in stageⅠ-Ⅱ(0.60±0.54) (P<0.05);The median survival time of patient with MMP-9 positive expressionwas 43.00±17.12 months and cumulate survival rate was 47.37% which significantly lower thanthat of MMP-9 negative expression (100%). In multivariate analyse, MMP-9, MMP-2, TIMP-2and residual tumor mass remained statistically independent prognostic factors. These results maysuggest the over-expression of MMP-9,MMP-2,MMP-7,TIMP-2,TIMP-3 may contribute to thedevelopment of ovarian tumors.The over-expression of MMP-9 and the imbalance betweenMMP-9 and TIMP-1 have significant relevance with the invasion and metastasis of ovariancancer.. MMP-9 would be as a prognostic indicator in patients with advanced ovarian cancer.In this study, the molecular cloning technique was used to construct the full length humanTIMP-1 cDNA eukaryotic expression vector and the DNA sequence was detected. The mutationof TIMP-1 was examined by used single strand conformation polymorphism(PCR-SSCP) in 40case of malignance and 15 case of benign ovarian tumor and 18 case of normal ovarian tissueswith the positive expression of TIMP-1. Sequence analysis showed that the eukaryoticexpression recombinant plasmid pcDNA4-TIMP1 contained the coding region of human fulllength TIMP-1 gene, and a missense mutation was found in sixth exon by compared with theGeneBank information; The result of SSCP showed that mutation were found in 7 case ofovarian malignant tumor tissues.The mutation rate was 16.7%. No mutation was found in benigntumor and normal ovarian tissues. These result suggest that there are gene mutation of TIMP-1in malignant ovarian tissues; Constructed recombinant plasmid may provide a basis for furtherstudy in the relationship between TIMP-1 with the invasion and metastasis of ovarian cancer.In this study , MMP-9 antisense and sense oligonucleotide were designed and modify byphosphorothioic acid, which were transfected mediated by Lipofectamine into ovarian cancercell line HO-8910PM which was expressed MMP-9 induced by fibronectin . RT-PCR,Westernblotting and Gelatin zymography were used to detected MMP-9 expression of mRNA andprotein and enzymatic active; Cell growth curve,Colony forming and Flow cytometer were usedto detected cell prolifera...