摘要
目的:通过检测基质金属蛋白酶抑制剂RECK和基质金属蛋白酶( MMP) 2、9及微血管密度(MVD)在子宫内膜异位症中的表达,研究其在子宫内膜异位症中表达的特点,探讨子宫内膜异位症中RECK与MMP- 2、MMP- 9及MVD表达的相关性。方法:应用链霉亲和素-生物素-辣根过氧化物酶法(SP法)对48例异位内膜组织, 31例正常内膜组织中RECK和MMP-2、MMP-9及MVD的表达情况进行检测。结果: MMP-2、MMP-9、RECK在异位内膜中的阳性表达率分别为:89.6%,79.2% ,43.8%,在正常内膜中阳性表达率为:35.5%,19.4%,87.1%,MMP-2、MMP-9在异位内膜中表达明显高于正常内膜,RECK在异位内膜中表达明显低于正常对照组,差异有显著性(P<0.05);RECK表达与MMP-2、MMP-9表达呈负相关,(r=-0.468、-0.320,P<0.05);MMP-2、MMP-9、RECK的表达与临床分期无明显相关性(P>0.05)。RECK蛋白在异位内膜中阳性表达率为43.8%,正常子宫内膜中表达阳性率为87.1%,两组比较有显著差异(P<0.05)。MVD计数在异位内膜表达为17.55±4.82,在正常内膜中为8.79±2.45,两组比较有显著差异(P<0.05)。MMP-2的表达与MVD呈正相关(r=0.783,P<0.05),MMP-9的表达与MVD呈正相关(R=0.637 P<0.05),RECK的表达与MVD呈负相关(r=-0.530,P<0.05)。结论:RECK蛋白表达异常与MMP-2、MMP-9相关,RECK蛋白的低表达有与EMs的新生血管生成有关,在EMs发生发展中起重要作用。
Background: Endometriosis (EMs) is that endometrial tissue (glands and stroma) which has the function of growing appear in anywhere but in the uterine cavity and myometrium. The disease has variety of clinical manifestations, although histology is benign, there are prolifera-tion, invasion, metastasis , recurrence and so on , such theses vicious acts. For child-bearing age women, it is one of the most common diseases. The pathogenesis of endometriosis has a variety of doctrines, but the most widely accepted theory is the blood current cultivation. Counter-current of menstruation is very widespread among child bearing period of women, the incidence is as high as 76% ~ 90%, but the incidence of the endometriosis is only 10% to 20%, it indicates that counter-current of menstruation may be only incentives, and the acting such as adhesion, erosion, proliferation and differentiation similar to the biological behavior of tumors of the ectopic endometrium to ovarian and other pelvic organs and peritoneal is the key to its pathogenesis. The cultivation , erosion and growth of endometrial debris reflux-ing into the abdominal cavity need extracellular matrix (ECM) destruction and reconstruction. Degradation of ECM depends mainly on the serine protease, cysteine protease, caspase and matrix metalloproteinase (matrix met-alloproteinases, MMPs), that 4 kinds of hydrolase-like protein, and MMPs are a class of the more important, almost can degrade all extracellular ma-trix components, and is closely related to the invasion and metastasis of tumor cells. The study in recent years found that EMs patients likely to be impacted by matrix metalloproteinases (MMPs) and their specific tissue inhibitor (TIMP).
Matrix metalloproteinases are a group of protease that can lead to the degradation of majority of extracellular matrix and various types of collagens,and are zinc- depen dent proteolytic enzymes, its activity determines the regulation of extracellular matrix degradation. When an error control mechanism of MMPs, extracellular matrix destructive degradation have been ex-cessive and too frequent, it would lead to the occurrence of many diseases and the deteriora-tion of the disease process (such as endometriosis, cancer, arthritis, etc.). Among them , MMP-2 and MMP-9 belong to gelatinases, mainly play a role with typeⅣcollagen as sub-strate, often exist in the form of non-activity, and in malignant tumors ,often be acti-vated,expressed highly .
RECK (reversion inducing cysteine rich protein with Kazal motifs) is a newly discovered tumor suppressor gene, it has a unique fuction of inhibiting expression and activity of matrix metalloproteinase (matrix matalloproteinases, MMPs), is a matrix metalloproteinase inhibitor. It is known by now that RECK gene at least suppress three kinds of MMP at the level of post-transcriptional: MMP-2, MMP-9 and MT1-MMP, thereby inhibit angiogenesis and me-tastasis of tumor further. Studies have shown that RECK gene expression in liver cancer, pan-creatic cancer, breast cancer, lung cancer was negatively correlated to tumor invasiv- eness,and patients with higher expression of RECK gene often have better prognosis significantly than patients with low expression. Therefore it is considered that the RECK gene is a new tumor suppressor gene, its mechanism may be related to inhibition of the wide range of MMPs.
Objective: To investigate the expression and clinical significance of RECK, MMP-2, MMP-9 and MVD in ectopic endometrium of the EMs.
Method: Immunohistochemical S-P was used to detect the expression of RECK, MMP-2, MMP-9 in 48 cases of ectopic endometrial tissue, 31 cases of normal endometrial tissue, at the same time to observe microvessel density (MVD) in different organizations .
The results: MMP-2, MMP-9, RECK were expressed mainly in the cytoplasm, rates of the positive expression in the ectopic endometrium respectively were: 89.6%, 79.2%, 43.8%, the positive expression rates in the normal endometrium were: 35.5 %, 19.4%, 87.1%, MMP-2, MMP-9 expression in ectopic endometrium was significantly higher than in normal endo-metrium, RECK expression in ectopic endometrium was significantly lower than the control group, there was a significant difference (P <0.05); RECK expression was negatively corre-lated to MMP-2, MMP-9 expression, (r =- 0.468, -0.320, P <0.05); MMP-2, MMP-9, RECK expression and clinical stage had no significant correlation (P> 0.05). MVD count in the ec-topic endometrium expressed as 17.55±4.82, in the normal endometrium for 8.79±2.45, compared the two groups were significantly different (P <0.05). MMP-2 expression and MVD was positively correlated (r = 0.783, P <0.05), MMP-9 expression and MVD was positively correlated too(R = 0.637 P <0.05), while RECK expression and MVD was a negative correla-tion (r =- 0.530 , P <0.05).
Conclusion:
1.In the EMs, RECK protein expression was significantly decreased, suggesting that in the ectopic endometrium the RECK gene expression was reduced, the lack of RECK promote the invasion of endometriotic cells,ability.
2.That MMP-2, MMP-9 protein expression in ectopic endometrium was significantly in-creased, and MMP-2 protein, MMP-9 protein expression and clinical stages of EMs had no significant correlation ,shows the severity of the disease has nothing to do with the ag-gression, the ectopic tissues in various stages of the disease have the same invasion.
3.Microvessel density (MVD) in ectopic endometrium was significantly higher in ectopic endometrium,it indicated that some factors led to the increasion of microvessel density (MVD). MMP-2, MMP-9 proteins in EMs with MVD count were positively correlated. It showed that the MMP-2, MMP-9 over-expression promoted the proliferation of blood vessels in EMs.
4.In ectopic endometrium, the RECK protein expression decreased , and were nega-tively correlated with MMP-2 protein, MMP-9 protein and microvessel density (MVD) expression ,suggesting that the increasing expression of RECK provides a new way of thinking as a treatment method of EMs and a new EMs treatment drug develop- ment,and offers a new target for the treatment of EMs.
引文
[1]SeliE,Berkkanoglu M,Arici A. Pathogenesis of endometriosis[J].Ob-stet Gynecol Clin North Am,2003,30(1):41-61.
[2]李全香,基质金属蛋白酶-2与子宫内膜异位症的相关研究[J].肿瘤防治杂志,2005,12(9):711-714.
[3]Ohbashi H.Matrix Metalloproteinases in Lung Diseases.Curr Protein PeptSci, 2002, 3 (4):409-421.
[4]谢祝英,韩雪松,基质金属蛋白酶及其抑制物在正常子宫内膜及子宫内膜病变中的作用[J].中华妇产科杂志,2004,39(10):711-713.
[5]高东梅,基质金属蛋白酶及抑制剂的研究进展[J]实用肿瘤杂志, 2004,(01):83-87.
[6]Hashimoto G,Inoki I, Fujii Y,et al.Matrix metalloproteinases cleave connective tissue growth factor and reactivate angiogenic activity of vascular endothelial growth factor 165[J].J Biol Chem,2002,277(39):36288-36295.
[7]Rigot V,Marbaix E,Lemoine P,et al.Invivo perimenstrual activation of progelatinase B(pro MMP-9)in the human endometrium and its dependence on stromelysin 1(MMP-3)ex vivo[J].Biochem J,2001,358(Pt1):275-280.
[8]TakahashiC,ShengZ,HoranTPetal.Regulation of matrixmetalloproteinase- 9 and inhib- ition of tumor invasion by the membrane–anchored glycoprotein RECK[J].Proc Natl Acad SciUSA,1998,95(22):13221 13226.
[9]Oh J,Takahashi R,Kondo S,et al.The membrane-anchored MMP inhibitor RECK is a key regulator of extracellar matrix integrity and angiogenesis.Cell,2001,107(6):789-800.
[10]Welm B,Mott J,Werb Z.Developmental biology:vasculogenesis is a wreck without RECK.Curr Biol,2002,12(6);209-211.
[11]Junseo Oh,Rei Takahashi,Shunya Kondo,et al.The membrane-anchored MMP inhibit or RECK is a key regulator of extracellular matrix integrity and angiogenesis[J]. Cell, 2001, 107(6):789-800.
[12] Nap AW,Dunselman GA,de Goeij AF,et al. Inhibiting MMP activity prevents the de-velopment of endometriosis in the chicken chorioallantoic membrane model. HumReprod, 2004,19:2180-2187.
[13]韩璐,董旭.子宫内膜异位症在位内膜和异位内膜的MMPs活性分析[J].中国实用妇科与产科杂志,2003,19(8):485-486.
[14]邱芳,高雪梅,罗国林,杨开选,王和,刘辉,文怡,基质金属蛋白酶及其抑制物与子宫内膜异位症关系的研究[J].四川大学学报(医学版) , 2006,37(1):118-122.
[15]李艳,郎景和,基质金属蛋白酶-9及其组织抑制剂-1在子宫内膜异位症组织中的表达[J].中华妇产科杂志, 2006, 41(01):30-33
[16]Hofmann UB,Westphal JR,Van Kraats AA,et al.Expression of integrin alpha(v)beta(3) correlates with activation of membrane-type matrix metalloproteinase-1(MT1-MMP) and ma-trix metalloproteinase-2(MMP-2)in human melanoma cells in vitro and in vivo.Int J Can-cer,2000,87:12-19.
[17]陈琼华曲军英基质金属蛋白酶-9及其抑制剂在异位和在位子宫内膜组织中的表达[J].中华妇产科杂志,2004,39(12):809-812.
[18]Welm B,Mott J,Werb Z.Developmental biology:vasculogenesis is a wreck without RECK.Crurr Biol,2002,12(6):209-211.
[19]Brew K,Dinakarpandian D,Nagase H.Tissue inhibitors of metalloproteinases: evolu- tion,structure and function.Biophys Acta,2000,1477(1-2):267-283.
[20]李喆;孙涛;魏雷震;王运杰;张朝东,RECK基因在脑胶质瘤细胞中的表达及其临床意义[J].中华神经外科杂志,2006,22(9):550-552.
[21]徐振宇,许传亮,孙颖浩. RECK基因在前列腺组织中的表达及其与基质金属蛋白酶-9的关系[J].中华实验外科杂志, 2005,22 (09):1146
[22]张勇,郑启昌,卢昕,张景辉,田元.肿瘤抑制基因RECK在胃癌中的表达及临床意义[J].肿瘤防治杂志, 2005,12 (02):110-113
[1]Seli E,Berkkanoglu M,A rici A..Pathogenesis of endometriosis[J].Obstet Gynecol Clin Eorth A m,2003,30(1):41-61.
[2]齐璇,基质金属蛋白酶及其抑制物与子宫内膜异位症[J].国外医学妇幼保健分册,2002, 13(1):14-16。
[3]Nap AW,Dunselman GA,de Goeij AF,et al. Inhibiting MMP activity prevents the de-velopment of endometriosis in the chicken chorioallantoic membrane model. Hum- Re-prod,2004,19:2180-2187.
[4]Koshikawa N,Giannelli G,Cirulli V,et al.Role of cell surface metalloproteinase MT1-MMP in epithelial cell migration over laminin-5[J].J Cell Biol,2000,148(3):615-624.
[5]Ivaska J,HeinoJ.Adhesion receptors and cell invasion:mechanisms of integrin-guided degradation of extracellular matrix [J].Cell Mol Life Sci,2000,57(1);16-24.
[6]Uzan C, Cortez A, Dufournet C, Fauvet R, Siffroi JP, Dara? E. Eutopic endometrium and peritoneal, ovarian and bowel endometriotic tissues express a different profile of matrix metalloproteinases-2,-3 and -11, and of tissue inhibitor metalloproteinases-1 and -2[J]. Virchows Arch. 2004 Dec;445(6):603-9. Epub 2004 Sep 28.
[7]韩璐,董旭.子宫内膜异位症在位内膜和异位内膜的MMPs活性分析[J].中国实用妇科与产科杂志,2003,19(8):485-486.
[8]Olson MW,Bemardo MN,Pietila M,et al.Characterization of the monomeric and dimeric forms of latent and active matrix metalloproteinase-9[J].J Biol Chem, 2000,275(4):2661-2668.
[9]OndoK,SugioK,Yamazaki K,et al.The significance of serum active matrix metallo- proteinase-9 in Patients with non-small cell lung cancer[J].Lung Cancer,2004,46(2):205-213.
[10]Edwards JG,MeLaren J,JonesJL,et al.Matrix metalloProteinases 2 and 9(gelatinases A and B)expression in malignant mesothelioma and benign Pleura[J].BrJ Cancer,2O03,88(10):1553-1559.
[11]Hofmann UB,Westphal JR,Van Kraats AA,et al.Expression of integrin alpha(v)beta(3) correlates with activation of membrane-type matrix metalloproteinase-1(MT1-MMP) and ma-trix metalloproteinase-2(MMP-2)in human melanoma cells in vitro and in vivo.Int J Can-cer,2000,87:12-19.
[12]李艳,郎景和,基质金属蛋白酶-9及其组织抑制剂-1在子宫内膜异位症组织中的表达[J].中华妇产科杂志, 2006, 41(01):30-33
[13]陈琼华曲军英基质金属蛋白酶-9及其抑制剂在异位和在位子宫内膜组织中的表达[J].中华妇产科杂志,2004,39(12):809-812.
[14] Zuker S, Cao,I Chen W T.Critical appraisal of the use of matrix metalloproteinases inhibitors in cancer treatment[J].Oncogenne,2000,19(56):6642-66550.
[15]Chung HW, Lee JY, Moon HS,et al. Matrix metalloproteinase-2, membranous type 1 matrix metalloproteinase, and tissue inhibitor of metalloproteinase-2 expression in ectopic and eutopic endometrium.Fertil steril,2002;78(4):787-795.
[16]Ria R,Loverro G,Vacca A et al.Angiogenesis extent and expression of matrix metal-loproteinase-2 and -9 agree with progression of ovarian endometriomas.Eur J Clin In-vest,2002,32(3):199-206
[17]Takenaka K,Ishikaw a S,Kawano Y,etal.Expression of a novel matrix metallo- pro-teinase regulator,RECK,and its clinical significance in resected non-small cell lung can-cer[J].Eur J Cancer,2004,40(10):1617-1623.
[18]李晟磊,刘宗文,赵秋民,于金霞,赵志华,高冬玲,庞霞,陈奎生,张云汉,食管鳞癌组织中RECK mRNA和蛋白的表达及意义[J].中国肿瘤临床,2007,34(22):1280-1286.
[19]张勇,郑启昌,卢昕,秦涛,转染RECK基因对肝癌细胞生物学行为的影响[J].中国普外基础与临床杂志,2006,13(2):150-153.
[20] Masui T,DoiR,Koshiba T, et al.RECK expression in Pancreatic cancer:its correlation with lower invasiveness and better Prognosis[J].Clin Cancer Res,2003,9(5):1779-1784.
[21]Junseo Oh,Rei Takahashi,Shunya Kondo,et al.The membrane-annhored MMP inhibi-tor RECK is a key regulator of extracellular matrix integrity and angiogenesis [J].Cell,2001,107(6):789-800.
[22]Xavier P,Beires J.Endometriosis:clinical aspects:Suband endometrial blood flow in patients with endometriosis[J].Hum Reprod, 2004,19:1168.