| Backgroud and purposeOvarian cancer is one of the three common gynecologic malignant tumor.In recent years,with the development of science and technology, detection technology of ovarian cancer has been rapidly improved.However,the ovary is in the depth of the pelvic cavity,in the early stage the occurence of ovarian cancer is difficult to be detected. Once there are obvious symptoms in the bodies,the patients are often in the late stage,they usually lose the best opportunities of surgical operaion.So,the mortality of ovarian cancer is the top of the gynecological tumors. We should find the biological indicators of ovarian cancer, explore the mechanism of occurrence and invasion of ovarian cancer. It is essential to improve the rate of early diagnosis of ovarian cancer and find out a good way to curb the recurrence and metastasis. Neutrophil gelatinase associated lipocalin (NGAL) is a novel protein,it is found in matrix metalloproteinase-9(MMP-9) in1993by Kjeldsen,etc. It is a new member of the Lipocalin family. NGAL is upregulated in inflammatory diseases and a variety of tumors. It is a new hot spot in meidical field in recent years. MMP-9can degrade the extra cellular matrix and basement membrane and affect the organization of tissue. Thereby it can promote tumor invasion and metastasis.A number of studies have shown that NGAL and MMP-9are closely related to each other.The expressions of NGAL and MMP-9in serum and ovarian tissue are detected by Reverse transcriptase-polymerase chain reaction (RT-PCR) and ELISA.We explore their correlation with clinicopathological characteristics and investigate the correlation between NGAL and MMP-9expressions in ovarian cancer.Materials and Methods1Experimental samples:We selected89patients who saw a doctor in the First Affiliated Hospital of Zhengzhou University during January2010to February2011.They were divided into three groups:50patients with epithelial ovarian cancers(31cases of serous carcinoma,19cases of mucinous carcinoma,according the International Federation of Gynecology and Obstetrics (FIGO) clinical staging standards:17cases of stage Ⅰ～Ⅱ,33cases of stage III-IV,patients aged38-72years old with a median age of53.5years old);26patients with benign ovarian tumors (15cases of the serous cystadenoma,6cases of the mucinous cystadenoma,patients aged28-67years old with a median age of48years old).Selected18cases of normal ovarian tissue and blood samples as control,they undertook the operation due to uterine fibroids during the same period in the hospital (patients aged43-61years old with a median age of47.5years old). The groups of these cases have not any significant differences in age(P>0.05).The selected cases had no the history of other tumors,they did not receive preoperative radiotherapy and chemotherapy, and they were confirmed by pathology after operation. We had told the patients everything about this test and they agreed on it. The patients were phlebotomized fasting venous blood4ml before operation. The blood specimens had been placed in the clean tubes without anticoagulant for30minutes in room temperature.Then they were centrifugalized for5minutes in4℃,3000r/min. We placed the supernatant fluid into EP tube and stored them in-80℃refrigerator. Got the parts of the ovarian tissue from the special patients during the operation and quickly moved them into-80℃refrigerator in0.5h, used to extract mRNA by semi-quantitative RT-PCR. Avoid repeated freezing and thawing for all specimens.2Experimental Methods:The expressions of NGAL and MMP-9in serum and ovarian tissue were detected in patients with epithelial ovarian cancer,the cases with ovarian benign tumor and the healthy women by Reverse transcriptase-polymerase chain reaction(RT-PCR) and ELISA. 3Statistical analysis:The SPSS17.0statistical software were used for data processing and analysis.The data was analyzed by t-test, K-S test and K-W test.Data correlation analysis applied Pearson analysis.a=0.05was deemed the inspection standard.Results1RT-PCR results:The expression levels of NGAL mRNA and MMP-9m RNA in the epithelial ovarian cancer tissue were (0.747±0.146,0.625±0.144) res pectively.Both of them in the epithelial ovarian cancer tissues were significantl y higher than in benign ovarian tumor tissues (0.442±0.155,0.312±0.064) and i n normal ovary tissues (0.121±0.037,0.111±0.042),the differences between them were statistically significant (all P<0.05).The expression levels of NGAL mRN A and MMP-9mRNA in the ovarian cancer tissues of stage Ⅰ～Ⅱ were (0.684±0.179,0.528±0.136) respectively, compared to the ovarian cancer tissues of st age Ⅲ-Ⅳ (0.779±0.116,0.676±0.124),the differences were statistically significa nt (P<0.05).The expression levels of NGAL mRNA and MMP-9mRNA in lym phnode metastasis group were (0.806±0.073,0.699±0.109),the expression levels o f the group without lymph node metastasis were (0.678±0.179,0.539±0.132),th e differences were statistically significant (all P<0.05).The expressions of NG-A L mRNA in the highly,moderately and poorly differentiated carcinoma were (0.824±0.065,0.770±0.064,0.620±0.221) respctively,the differences were statistically significant (all P<0.05),and the expressions of MMP-9mRNA in the highly,mo derately and poorly differentiated carcinoma were (0.505±0.142,0.635±0.111,0.715±0.120) respctively,the differences were statistically significant (all P<0.05).2ELISA results:The NGAL and MMP-9serum levels of patients with epithelial ovarian cancer were (44.799±8.723,39.925±7.455) ng/ml respectively.Both in the serum of patients with epithelial ovarian cancer were significantly higher than the patients with benign ovarian tumors (31.944±4.190,35.321±8.759) ng/ml and normal control group (17.787±2.655,24.176±3.405) ng/ml,the differences were statistically significant (all P<0.05).The serum levels of NGAL and MMP-9in the stage Ⅰ-Ⅱ of ovarian cancer patients respectively were (40.434±7.642,36.834±6.988) ng/ml, compared to the serum levels in the stage Ⅲ～Ⅳ of ovarian cancer patients (47.048 ±8.484,41.519±7.279) ng/ml,the differences were statistically significant (all P<0.05).The serum levels of NGAL and MMP-9in lymph node metastasis group were (47.280±8.523,41.537±7.273) ng/ml,the serum levels of the group without lymph node metastasis were (39.985±7.110,36.799±6.9761) ng/ml,the differences were statistically significant (all P<0.05).The serum levels of NGAL in the highly,moderately and poorly differentiated carcinoma respectively were (48.535±8.520,44.907±9.611,40.319±5.125) ng/ml,the differences were statistically significant (all P<0.05),and the levels of MMP-9in the highly, moderately and poorly differentiated carcinoma respectively were (37.625±7.197,38.639±7.748,43.808±6.033) ng/ml, the differences were statistically significant (all P<0.05).3correlation analysis:We found that the level of NGAL mRNA in the epithelial ovarian cancer tissue and the serum level of NGAL of patients with epithelial ovarian cancer were positively correlated (r=0.762,P<0.05) using Pearson correlation analysis, the level of MMP-9mRNA in the epithelial ovarian cancer tissue and the serum level of MMP-9of patients with epithelial ovarian cancer were positively correlated (r=0.601,P<0.05),the expression level of NGAL mRNA and MMP-9mRNA in the epithelial ovarian cancer tissue were positively correlated (r=0.740,P<0.05), the NGAL and MMP-9serum levels of patients with epithelial ovarian cancer were positively correlated (r=0.676,P<0.05).Conclusion1The up-regulated expressions of NGAL and MMP-9are closely associated with the tumor malignancy biological behaviour of ovarian epithelial cancer.2NGAL and MMP-9may play an important synergistic action in the genesis development and metastasis of ovarian cancer.3Detecting serum levels of NGAL and MMP-9may be of great significance for the differential diagnosis of benign and malignant tumors. |
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