子宫内膜异位症腺上皮细胞中NF-кBp65干预OPN调节MMP9表达及侵袭性研究

发布时间：2018-01-28 01:33

本文关键词： 子宫内膜异位症核转录因子-κBp65 骨桥蛋白金属基质蛋白酶-9 细胞侵袭性　出处：《宁夏医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的1.明确子宫内膜异位症在位内膜腺上皮细胞中NF-кBp65、OPN及MMP9表达情况及其相关性。2.分析子宫内膜异位症在位内膜腺上皮细胞中核因子NF-кBp65在OPN调节MMP9表达中的作用。3.探讨子宫内膜异位症在位内膜腺上皮细胞中NF-κBp65干预前后细胞侵袭性的变化。方法1.收集分泌期EMs患者在位内膜12例,将EMs腺上皮细胞进行原代分离、培养、纯化,通过免疫细胞化学法测定OPN、NF-кBp65及MMP9在腺上皮细胞中的表达情况;分别通过基因沉默及质粒导入的方法,对内异症患者分泌期在位内膜腺上皮细胞中NF-кBp65进行干预,Real Time-PCR及western-blot检测NF-кBp65、OPN、MMP9 mRNA及蛋白表达的变化。2.通过侵袭小室实验测定干预NF-кBp65基因前后细胞侵袭性的变化。结果1.免疫细胞化学结果显示:NF-кBp65、OPN及MMP9主要定位于EMs腺上皮细胞的胞浆中。2.NF-кBp65 siRNA基因沉默后腺上皮细胞中NF-кBp65、MMP9 mRNA及蛋白的表达较干预前明显下降,其差异具有统计学意义(P0.05),NF-кBp65质粒导入后NF-кBp65及MMP9 mRNA及蛋白表达均明显增高,其差异具有统计学意义(P0.05),NF-кBp65 siRNA基因沉默及质粒导入后OPN表达均无明显改变,差异无统计学意义(P 0.05)。3.内异症患者在位内膜腺上皮细胞中,NF-кBp65 siRNA干预后其细胞侵袭性较干预前明显降低,NF-кBp65质粒干预后其细胞侵袭性明显增加,其差异具有统计学意义(P均0.05)结论1.NF-кBp65 siRNA沉默EMs患者在位内膜腺上皮细胞后,NF-кBp65及MMP9的表达明显下降;NF-кBp65质粒上调EMs患者在位内膜腺上皮细胞后,NF-кBp65及MMP9的表达明显增高,揭示了NF-кB是干预OPN调节MMP9在内异症患者内膜中异常表达的关键。2.NF-кBp65 siRNA基因沉默EMs患者在位内膜腺上皮细胞后,腺上皮细胞的侵袭性明显降低;NF-кBp65质粒上调EMs患者在位内膜腺上皮细胞后,腺上皮细胞的侵袭性明显增加,提示内异症中NF-кBp65通过调节OPN/MMP9表达改变细胞侵袭性。
[Abstract]:Objective 1. To identify NF- Bp65 in eutopic endometrial glandular epithelial cells in endometriosis. Expression of OPN and MMP9 and their correlation. 2. Analysis of the role of nuclear factor NF- Bp65 in the regulation of MMP9 expression by OPN in eutopic endometrial glandular epithelial cells in patients with endometriosis. Objective: to investigate the changes of invasiveness of eutopic endometrium in eutopic endometrial glandular epithelium before and after intervention of NF- 魏 Bp65. Methods 1. 12 cases of eutopic endometrium in secretory phase of EMs were collected. EMs glandular epithelial cells were isolated, cultured and purified. The expression of Bp65 and MMP9 in glandular epithelial cells was determined by immunocytochemistry. By gene silencing and plasmid transfection, NF- Bp65 in eutopic endometrial glandular epithelial cells in the secretory phase of patients with endometriosis was interfered with. Real Time-PCR and western-blot were used to detect NF- Bp65 and OPN. Changes of MMP9 mRNA and protein expression. 2. The changes of cell invasiveness before and after intervention of NF- Bp65 gene were determined by invasive chamber assay. Results 1. Immunocytochemistry showed that: 1. NF- Bp65. OPN and MMP9 were mainly located in the cytoplasm of EMs glandular epithelial cells. 2. NF- Bp65 in glandular epithelial cells after Bp65 siRNA gene silencing. The expression of MMP9 mRNA and protein was significantly lower than that before intervention, and the difference was statistically significant (P 0.05). The expression of NF- Bp65 and MMP9 mRNA and protein were significantly increased after the introduction of NF- Bp65 plasmid, and the difference was statistically significant (P0.05). The silencing of NF- Bp65 siRNA gene and the expression of OPN in plasmid were not significantly changed. There was no significant difference (P < 0.05). The invasiveness of Bp65 siRNA in eutopic endometrial glandular epithelial cells was significantly lower than that before intervention. The invasion of NF- Bp65 plasmid was significantly increased after intervention. The difference was statistically significant (P 0.05). Conclusions 1.After silencing EMs patients with Bp65 siRNA, the eutopic endometrial glandular epithelial cells were silenced. 2. The expression of Bp65 and MMP9 decreased significantly. Bp65 plasmid upregulated the expression of Bp65 and MMP9 in eutopic endometrial glandular epithelial cells of EMs patients. It is revealed that NF- MMP9 B is the key to interfere with OPN to regulate abnormal expression of MMP9 in endometrium of patients with MMP9. 2. NF- Bp65. The siRNA gene silenced the glandular epithelial cells of the eutopic endometrium of EMs patients. The invasiveness of glandular epithelial cells was significantly decreased. The invasiveness of eutopic endometrial glandular epithelial cells in patients with EMs was significantly increased after NF- Bp65 plasmid upregulated the invasiveness of glandular epithelial cells. The results suggest that NF- Bp65 changes the invasiveness by regulating the expression of OPN/MMP9 in endometriosis.
【学位授予单位】：宁夏医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R711.71

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