Detection And Significance Of SDC-1,HLA-G And MMP-9in Endometriosis

Background and ObjectiveEndometriosis (endometriosis, EMS) is a benign disease of gynaecology,whichrefers to the endometrial glands and stroma planting outside the uterine cavity. Itspathogenesis remains unclear, EMS is nature of the benign and the behavior is similarto malignant tumor,which showed adjacent organs plantation and destruction, thesurrounding tissue invasion, distant parts of the transfer, unlimited growth, treatmentmore difficult thoroughly, etal.Then,which were affecting tumor cells fromdegradation, migration, adhesion, proliferation and infiltration process of variousmolecules are widely used in the study of EMS.Syndecan-1(syndecan-1, SDC-1)belongs to the members of the family ofheparansulfateproteoglycan, which is a classic growth factor,angiogenic factors,chemokines, angiogenesis factor, chemokines, its heparan sulfate chain has extensivestructural heterogeneity, which can combined with extracellular matrix(extracellularmatrix, ECM), cell surface adhesion molecules, growth factors, enzymes, etal,involved in cell proliferation, adhesion, migration and angiogenesis process.Humanleucocyteantigen-G(humanleucocyteantigen-G, HLA-G) is a non-classicalMHC-Ib antigen molecules, by inhibiting the activity of NK cells and T cells thatparticipate in the immune escape of tumor cells.The degradation of the ECM is EMS ectopic endometrial cells invasion and the main way to plant, and MMPs play animportant role in the degradation of the ECM. Matrix metalloproteinase-9(matrixmetalloproteinase-9, MMP-9) can degrade the ECM, penetrate the cell of thebasement membrane, through interaction with integrin, to strengthen cell-celladhesion effect, at the same time to promote the vascular endothelial cell budding,conducive to the formation of new blood vessels.It has not been reported about the joint detection of SDC-1,HLA-G and MMP-9in EMS patients domestic and overseas. In this experiment,the SDC-1,HLA-G andMMP-9in the ectopic endometrium、eutopic endometrium and in serum levels ofEMS patients are detected by immunohistochemical S-P method (immunohisto-chemical S-P method) and enzyme linked immune sorbent assay(enzyme linkedimmune sorbent assay, ELISA);Then, analyze their expressions in different clinicalstage in EMS and their correlationships with eachother, aiming to investigate the roleof SDC-1,HLA-G and MMP-9in the progression of EMS, and provides theoreticalbasis on the pathogenesis and clinical studies for EMS.Materials and methods1. Research object1.188patients with “ovarian endometriosis” were selected from July2012toSeptember2013in the Department of Gynecology from our hospital, which werecomfromed for “ovarian endometriosis” after laparoscopic surgery and postoperativepathology. American fertility association revised standard (R-AFS) in1985was usedto divide the clinical stage:Ⅰ-Ⅱ stage44cases, Ⅲ-Ⅳ stage44cases; The controlgroup was selected35cases in patients with septum uterus who were givenlaparoscopic-hysteroscopic surgery in the same period.1.288cases ovarian endometriosis specimens were took HE staining,60caseswere selected in the test:Ⅰ-Ⅱ stage30cases, Ⅲ-Ⅳ stage30cases; Collect aspecimen of ovarian chocolate cyst as part of the capsule wall as the group ectopicendometrium;Among60patients,48cases who had fertility requirements weretreated by the hysteroscopy operation at the same time,excepted the endometriallesions, took “eutopic endometrium”as the group eutopic endometrium.35cases inpatients with septum uterus who were given laparoscopic-hysteroscopic surgery, excepted the endometrial lesions and EMS,took “normal endometrium” as the controlgroup.1.3123cases fasting elbow venous blood was taken in the early morning priorto surgery.1.4Inclusion criteria:(1) Women of childbearing age, preoperative menstrualcycle is normal;(2)3months did not use hormonal drugs;(3) No other diseases,without a history of malignant tumors;(4) No acute pelvic inflammation and otherpelvic changes;(5) The specimens for patients in the menstrual cycle time for theproliferation of period.2. Experimental methods2.1The expressions levels of SDC-1,HLA-G and MMP-9with EMS patientsand the control group were detected by Immunohistochemical S-P method.2.2The serum expression levels of SDC-1,HLA-G and MMP-9in EMS patientsand the control group were detected by ELISA.3. Statistical methodsAll data analysised by statistical software SPSS17.0. One-way ANOVA usedfor Multiple sets of quantitative data, LSD test used for multiple comparison,Independent-samples t test used for two groups of independent sample comparison,Pearson product-moment correlation coefficient used for Continuous random variablecorrelation analysis. The ROC curve analysis used for the sensitivity and specificity.Inspection level of α=0.05.Results1. The expression levels of SDC-1,HLA-G and MMP-9in EMS group and thecontrol group in the tissuesSDC-1、HLA-G and MMP-9protein mainly expressed in the cytoplasm ofgland epithelial cells in ectopic endometrium and eutopic endometrium in patientswith EMS,and little expressed in the cytoplasm of stromal cells.In group ectopic endometrium、eutopic endometrium and the control group, theexpression levels of SDC-1were146.35±19.56,114.86±13.67and70.30±11.23，(F=277.44,P=0.000);the expression levels of HLA-G were152.69±16.08,129.06±8.93and61.95±9.40，(F=654.84,P=0.000);the expression levels of MMP-9were148.11 ±22.88,117.57±8.72and83.98±9.74，(F=188.97,P=0.000). Comparison betweenany two means,the expression levels of SDC-1,HLA-G and MMP-9protein in groupgroup ectopic endometrium and eutopic endometrium were much higher than that inthe control group(P <0.05). The expression levels of SDC-1,HLA-G and MMP-9protein in group group ectopic endometrium were much higher compared to that ingroup eutopic endometrium (P <0.05).In group ectopic endometrium with EMS,the expression levels of SDC-1,HLA-G and MMP-9protein at stageⅠ-Ⅱ were129.33±5.08,139.83±7.38and130.85±15.21; and at stage Ⅲ-Ⅳ their expression levels were163.37±12.39,165.55±11.39and165.36±14.76. In group eutopic endometrium, the expression levels of SDC-1,HLA-G and MMP-9protein at stage Ⅰ-Ⅱ of EMS were103.39±8.02,122.64±4.03and110.61±6.67, and at stage Ⅲ-Ⅳ their expression levels were126.33±6.57,135.49±7.77and124.54±3.10.In group ectopic endometrium and eutopic endometriu-m, the expression levels of SDC-1,HLA-G and MMP-9protein at stageⅠ-Ⅱwerelower than that at stage Ⅲ-Ⅳ(P <0.05).2. The serum expression levels of SDC-1,HLA-G and MMP-9in EMS groupand the control groupIn EMS group,the expression levels of SDC-1,HLA-G and MMP-9were3888.34±607.38μg/L,3888.34±607.38μg/L and15.42±2.61μg/L;In the control group,the expression levels of SDC-1,HLA-G and MMP-9were1481.91±482.49μg/L,5.77±1.82μg/L and9.01±3.15μg/L;The serum expression levels of SDC-1,HLA-Gand MMP-9in EMS patients were significantly higher than the control group(P<0.05);In group EMS,the expression levels of SDC-1,HLA-G and MMP-9protein atstage Ⅰ-Ⅱ were3463.20±352.24μg/L,9.98±2.30μg/L and14.84±2.871μg/L; and atstage Ⅲ-Ⅳ their expression levels were4313.48±502.44μg/L,11.50±1.54μg/L and16.01±2.20μg/L.The serum levels of SDC-1,HLA-G and MMP-9in stage III-IV ofEMS group were significantly higher than stage I-II (P<0.05);3. After the ROC curve analysis, the combined detection of SDC-1,HLA-G andMMP-9in the serum of EMS patients,the sensitivity and specificity were93.18%and 94.28%.The detection value were significantly higher than the detection of separateor in pairs to diagnosis of EMS.4. The correlations of SDC-1,HLA-G and MMP-9in EMS group and the controlgroupThe serum expression levels of SDC-1and HLA-G had positive correlation withthat of MMP-9in EMS patients(r=0.683, P<0.05;r=0.456, P<0.05); The serumexpression levels of SDC-1had no correlation with that of HLA-G in EMSpatients(r=0.212, P＞0.05).In ectopic endometrium with EMS group, the expression levels of SDC-1andHLA-G had positive correlation with that of MMP-9(r=0.338,P=0.008；r=0.302,P=0.019);the expression levels of SDC-1had no correlation with that of HLA-G(r=0.185, P=0.157).In eutopic endometrium with EMS group, the expression levels of SDC-1andHLA-G had positive correlation with that of MMP-9(r=0.476,P=0.002;r=0.601,P=0.000);the expression levels of SDC-1had no correlation with that of HLA-G(r=0.161, P=0.273).In the control group, the expression levels of SDC-1,HLA-G and MMP-9hadno positive correlation with eachother(r=0.067, P=0.651; r=0.052, P=0.752; r=0.107,P=0.512).Conclusion1.SDC-1,HLA-G and MMP-9may be involved in the progression of EMS;2.The combined detection of SDC-1, HLA-G and MMP-9had certain referencevalue for noninvasive diagnosis of EMS;3.SDC-1, HLA-G may take synergetic effects with MMP-9in the progression ofEMS.