本文關(guān)鍵詞： 子宮內(nèi)膜異位癥 核轉(zhuǎn)錄因子-κBp65 骨橋蛋白 金屬基質(zhì)蛋白酶-9 細胞侵襲性　出處：《寧夏醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的1.明確子宮內(nèi)膜異位癥在位內(nèi)膜腺上皮細胞中NF-кBp65、OPN及MMP9表達情況及其相關(guān)性。2.分析子宮內(nèi)膜異位癥在位內(nèi)膜腺上皮細胞中核因子NF-кBp65在OPN調(diào)節(jié)MMP9表達中的作用。3.探討子宮內(nèi)膜異位癥在位內(nèi)膜腺上皮細胞中NF-κBp65干預(yù)前后細胞侵襲性的變化。方法1.收集分泌期EMs患者在位內(nèi)膜12例,將EMs腺上皮細胞進行原代分離、培養(yǎng)、純化,通過免疫細胞化學(xué)法測定OPN、NF-кBp65及MMP9在腺上皮細胞中的表達情況;分別通過基因沉默及質(zhì)粒導(dǎo)入的方法,對內(nèi)異癥患者分泌期在位內(nèi)膜腺上皮細胞中NF-кBp65進行干預(yù),Real Time-PCR及western-blot檢測NF-кBp65、OPN、MMP9 mRNA及蛋白表達的變化。2.通過侵襲小室實驗測定干預(yù)NF-кBp65基因前后細胞侵襲性的變化。結(jié)果1.免疫細胞化學(xué)結(jié)果顯示:NF-кBp65、OPN及MMP9主要定位于EMs腺上皮細胞的胞漿中。2.NF-кBp65 siRNA基因沉默后腺上皮細胞中NF-кBp65、MMP9 mRNA及蛋白的表達較干預(yù)前明顯下降,其差異具有統(tǒng)計學(xué)意義(P0.05),NF-кBp65質(zhì)粒導(dǎo)入后NF-кBp65及MMP9 mRNA及蛋白表達均明顯增高,其差異具有統(tǒng)計學(xué)意義(P0.05),NF-кBp65 siRNA基因沉默及質(zhì)粒導(dǎo)入后OPN表達均無明顯改變,差異無統(tǒng)計學(xué)意義(P 0.05)。3.內(nèi)異癥患者在位內(nèi)膜腺上皮細胞中,NF-кBp65 siRNA干預(yù)后其細胞侵襲性較干預(yù)前明顯降低,NF-кBp65質(zhì)粒干預(yù)后其細胞侵襲性明顯增加,其差異具有統(tǒng)計學(xué)意義(P均0.05)結(jié)論1.NF-кBp65 siRNA沉默EMs患者在位內(nèi)膜腺上皮細胞后,NF-кBp65及MMP9的表達明顯下降;NF-кBp65質(zhì)粒上調(diào)EMs患者在位內(nèi)膜腺上皮細胞后,NF-кBp65及MMP9的表達明顯增高,揭示了NF-кB是干預(yù)OPN調(diào)節(jié)MMP9在內(nèi)異癥患者內(nèi)膜中異常表達的關(guān)鍵。2.NF-кBp65 siRNA基因沉默EMs患者在位內(nèi)膜腺上皮細胞后,腺上皮細胞的侵襲性明顯降低;NF-кBp65質(zhì)粒上調(diào)EMs患者在位內(nèi)膜腺上皮細胞后,腺上皮細胞的侵襲性明顯增加,提示內(nèi)異癥中NF-кBp65通過調(diào)節(jié)OPN/MMP9表達改變細胞侵襲性。
[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.
【學(xué)位授予單位】：寧夏醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R711.71

【參考文獻】

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1 李望舒;白治苗;王瑞;盧玉鳳;陳華;姚衛(wèi)衛(wèi);哈春芳;;NF-κB p65對子宮內(nèi)膜異位癥腺上皮細胞OPN、MMP-9表達及細胞侵襲性的影響[J];現(xiàn)代婦產(chǎn)科進展;2016年07期

，

本文編號：1469390