Screening Of Differentially Expressed Genes Of Endometriosis By CDNA Microarray Technique And The Investigation Of COX-2,VEGF

AIM: The etiology of endometriosis has remained enigmatic. The angiogenese theory becomes key one, the implateation and growth of ectopic endometrium is closely related with angiogenesis , angiogenesis plays important roles in endometriosis.So,we identified of differentially expressed genes involved in endometriosis by cDNA microarray technique and effects of COX-2 and VEGF which were related with angiogenese in order to reveal the pathogenesis of endometriosis.METHOD: 1. There are 1200 genes in gene chip,which marks by isotope.The gene expression pattern analysis of samples of endometriosis and normal control endometria tissues were detected by cDNA microarray technique.2.(l) The expression of COX-2 and VEGF in 32 cases of endometriosis patients were detected by immunohistochemistry methods,and analysed their (COX-2 and VEGF)correlation with clinicalbiological behavior. (2)Endometrium specimens were obtained from 30 in endometriosis and 29 normal uterus for control. Expressions of COX-2 mRNA were examined byin situ hybridization technique with specific cDNA probes.3.The 10 endometrium cells culture in vitro and the expression of COX-2 in ectopic endometrium cell by immunofluorescence technology by confocal laser microscope.RESULTS: 1. 15 cytokines genes expression were up-regulated among 197 differently expressed genes in 3 cases endometriosis tissues and in 3 cases normal control endometria tissues, include IL-1 IL-2, IL-6, IL-8, VEGF TGF, EGF FGF and EPO which is found recently ; COX-2 enzyme genes expression was noticeably up-regulated;in addition there was PGE2.2.(1) Expressions of COX-2 and VEGF in endometriosis were 84.3% and 93.8% , which were significant higher compared with normal endometriosis (P<0.001 ).COX-2 and VEGF of eutopic and ectopic endometrium in proliferative phase and secretory phase were 83.3%,85.7% and 94.4%,92.9%. The expression of COX-2 was closely correlate with VEGF (r=0.747, P<0.001).(2)Expression of mRNA of COX-2 in endometriosis were [proliferative phase (3.11 +0.56) , secretory phase (3.05 +0.41) , which were significant higher compared with normal endometriosis (P<0.001).There was no difference between proliferative and secretory endometrium of endometriosis.3.Cultured normal and ectopic endometrial cells in vitro:(l) There was no difference in morphology by optical microscope and electron microscope;(2) The expression of COX-2 was higher in ectopic than in normal endometrial cells by confocal laser microscope. CONCLUSION: l.cDNA microarray technique is effective in screening the genes in relative to endometriosis, which screens 17 up-regulational expression genes,including cytokin^ COX-2PGE.2.There are some expressions of COX-2. VEGF protein and mRNA in endometrium tissues 3.The expressions of COX-2 and VEGF protein and mRNA are higher in endometiosis. 4.There are no different expressions of COX-2 and VEGF in endometriosis between proliferative phase and secretory phase,meanwhile the expression of COX-2 is related with VEGF. 5. The increase expressions of COX-2 VEGF are associated with angiogenesis.