| OBJECTIVEThis research will explore whether epithelial-mesenchymal transition involved in the growth of endometrium outside the uterus, analysis the expression of mi R-200 a, mi R-200 b in ovarian endometriosis and the relationship with EMT. Consequently, it may offer a theoretical foundation in molecular level for revealing the mechanisms of invasion and metastasis in endometriosis. METHODS 1 Specimens collection: case group: eighty ectopic endometrium(ECs) and eutopic endometrium(EUs) of patients who underwent surgical treatment for ovarian endometriosis in the Tian Jin Central Hospital of Gynecology and Obstetrics from May 2014 to July 2014 were recruited and their average age is 36.74±7.56. All of them were pathologically confirmed endometriosis postoperatively. According to the 1997 revised classification of the American Society of Reproductive Medicine(r-ASRM) staging, I-II phase of the 21 cases, 30 III period, 29 of IV period.Control group: forty-three normal endometrium(ENs) of patients who underwent surgery due to simple ovarian cysts in the same hospital at the same time, their average age is 38.57±5.02. All of them had pathologically confirming normal endometrial tissue, without malignant transformation and endometriosis.All the samples had a normal menstrual cycle without a history of malignant tumors or other serious disease, and at least six months did not use hormonal drugs. 2 Experemental procedure 2.1 Specimen treatment: Immersed all specimens in sterile tube immediately and stored in liquid N2 until further process. 2.2 RNA and protein extraction: specimens of 50-100 mg in control group and case group should be grinded in liquid N2 for RNA and protein extraction respectively. 2.3 RT-PCR was used to detect the expression of mi RNA-200 a, mi RNA-200 b in each group. 2.4 Western Blot was used to detect the expression of EMT related proteins: ZEB1, ZEB2, E-cadherin, N-cadherin, Snail, Slug in each endometrium tissues. 2.5 Analysis the experimental data with SPSS software package. RESULTS 1.The expression of mi R-200a/b in case group and control groupMi R-200a/b in each tissue has different degree expression. Compared with ENs tissue, the expression level of mi R-200a/b in ECs and EUs were decreased obviously. Moreover, the expression of mi R-200a/b has a significant difference between ECs and EUs endometrium. 2.The expression of EMT related proteins in case group and control groupZEB1/2, E-cad, N-cad, Snail, Slug in each tissue had different degree expression. Compared with ENs tissue, the expression level of E-cad in ECs and EUs were decreased obviously, and it was significantly lower in ECs than EUs. On the contrary, the expressions of ZEB1/2, N-cad, Snail and Slug increased significantly, and there were significantly higher in ECs than EUs. 3.The comparison of correlation between mi R-200a/b expression rate and E-cad, ZEB1/2 expression rateAccording to Pearson Correlation Coefficients test, the expressions of mi R-200a/b had a significantly positive correlation with E-cad; meanwhile, mi R-200a/b had a significantly negative correlation with ZEB1/2. 4.The correlation between mi R-200a/b and endometriosis r ASRM stageAccording to the statistical results, the relative expression level of mi R-200a/b in ECs and EUs had no significant correlation with r ASRM stage or r ASRM score. CONCLUSION 1.The expression of mi R-200a/b and ZEB1/2 were related with endometriosis, which suggests that they may be involved in the genesis and progression of EMs and play important roles in the process of invasion and metastasis. 2. The strong correlation found between mi R-200a/b and ZEB1/2 indicates that the ZEB1/2 may be the target of mi R-200a/b. Both of them may participate in the progress of EMT in endometriosis. 3. The expressions of mi R-200a/b are obviously decreased in eutopic endometrium, aberrant gene expression in eutopic endometrium is a key point in the development of endometriosis, it will provide important clues for endometriosis early diagnosis, effective treatment and prognosis assessment. |
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