Results
| PMID | 26675577 |
| Gene Name | PTGS2 |
| Condition | Endometriosis |
| Association |
Associated |
| Population size | 40 |
| Population details | 40 (30 women with endometriosis, 10 controls without endometriosis) |
| Sex | Female |
| Other associated phenotypes |
Endometriosis |
|
Zhonghua Fu Chan Ke Za Zhi. 2015 Oct;50(10):770-6. Ma, Yange| Shen, Airong| Li, Canyu| Xu, Shengnan| Guo, Huanhuan| Zou, Shunhong Department of Obstetrics and Gynecology, The Third Affiliated Hospital, Zhengzhou University, Zhengzhou 450052, China.| Department of Obstetrics and Gynecology, The Third Affiliated Hospital, Zhengzhou University, Zhengzhou 450052, China; Email: shenai OBJECTIVE: To investigate the effect of targeted interruption of cyclooxygenase-2 (COX-2) gene by small interference RNA (siRNA) on the expression of COX-2, vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) in eutopic and ectopic endometrial stromal cells (ESC) with endometriosis, and the effect on the content of 6-keto-prostaglandin-F1alpha (6-keto-PGF1alpha, metabolites of COX) and the apoptosis of eutopic and ectopic ESC with endometriosis. METHODS: Ectopic and eutopic ESC from 30 women with endometriosis were isolated and cultured respectively. Then, ESC were classified into three groups: interference group, negative control group and blank control group. ESC in interference group were injected into siRNA transfection complex while ESC in negative control group were injected into negative control transfection complex. ESC from 10 participants without endometriosis were the normal control group. The mRNA and protein expression of COX-2, VEGF, MMP-9 in pre-transfected and post-transfected eutopic and ectopic ESC were detected through real time reverse transcription PCR and western blot. The content of 6-keto-PGF1alpha was determined by ELISA, the apoptotic cells were detected by flow cytometry. RESULTS: After interruption of COX-2 gene, there were no significant difference in the mRNA and protein expression of COX-2, VEGF and MMP-9 between the negative control group and blank control group (P > 0.05); the mRNA and protein expression of the three genes in interference group were significantly lower than those in negative control group and blank control group (P < 0.05); the mRNA expression of the three genes in interference group of eutopic ESC were 0.87 +/- 0.06, 1.76 +/- 0.59, 1.04 +/- 0.32, in interference group of ectopic ESC were 0.75 +/- 0.12, 1.62 +/- 0.47, 0.88 +/- 0.25, the protein expression of the three genes in interference group of eutopic ESC were 0.457 +/- 0.019, 0.500 +/- 0.012, 0.361 +/- 0.008, in interference group of ectopic ESC were 0.323 +/- 0.018, 0.474 +/- 0.016, 0.339 +/- 0.009; the mRNA and protein expression of the three genes in ectopic ESC had a more reduction than those in eutopic ESC (P < 0.05). The results from ELISA revealed that the content of 6-keto-PGF1alpha in the normal control group [(17.7 +/- 1.9) pg/ml] were significantly lower than those in the blank control group (P < 0.05), the content of 6-keto-PGF1alpha in ectopic ESC were significantly higher than that in eutopic ESC (P < 0.05), the content of 6-keto-PGF1alpha in the blank control group of eutopic and ectopic ESC were (32.4 +/- 2.6) pg/ml, (38.2 +/- 3.7) pg/ml; there was no significant difference in the content of 6-keto-PGF1alpha between the negative control group and blank control group (P > 0.05); compared with those of negative control group and blank control group, the content of 6-keto-PGF1alpha in interference group decreased significantly (P < 0.05), the content of 6-keto-PGF1alpha in interference group of eutopic and ectopic ESC were (17.1 +/- 2.4) pg/ml, (20.9 +/- 2.7) pg/ml; the content of 6-keto-PGF1alpha in eutopic ESC had a slightly more reduction than that in ectopic ESC (P > 0.05). The results from flow cytometry displayed that, there was no significant difference in apoptotic cells between the negative control group and blank control group (P > 0.05); compared with those of negative control group and blank control group, more apoptotic cells were detected in interference group and the difference was significant (P < 0.01); the apoptotic cells in ectopic ESC were significantly more than that in eutopic ESC (P < 0.05); the apoptosis rate in interference group of eutopic and ectopic ESC were (33.76 +/- 0.06)%, (47.18 +/- 0.12)%. CONCLUSIONS: Our results suggested the targeted interruption of COX-2 gene by siRNA effectively inhibited the mRNA and protein expression of COX-2, VEGF and MMP-9 in both eutopic ESC and ectopic ESC with endometriosis, greatly increased the apoptotic rate of cells and obviously reduced the content of 6-keto-PGF1alpha by inhibiting the activity of COX-2. And the changes in ectopic endometrium were more evident than those in eutopic endometrium. Mesh Terms: 6-Ketoprostaglandin F1 alpha| Apoptosis/*genetics| Cyclooxygenase 2/*genetics/metabolism| Endometriosis/*genetics/metabolism/pathology| Endometrium/drug effects/metabolism/pathology| Epithelial Cells| Female| Humans| Matrix Metalloproteinase 9/*me |
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