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  本文關(guān)鍵詞： SD大鼠子宮肌瘤模型 子宮平滑肌細(xì)胞的分化和增殖 子宮肌瘤 myocardin ERα　出處：《武漢科技大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：目的： 1.研究Myocardin誘導(dǎo)的子宮平滑肌細(xì)胞分化標(biāo)志基因的表達(dá) 2.研究ERα促進(jìn)子宮平滑肌細(xì)胞的增值作用 3.研究ERα所抑制的myocardin誘導(dǎo)的子宮平滑肌細(xì)胞分化標(biāo)志基因的轉(zhuǎn)錄激活和表達(dá) 4.研究ERα和myocardin之間的蛋白蛋白相互作用 方法： 1.建立SD大鼠子宮肌瘤模型。 2. SD大鼠子宮肌瘤模型建立后分別取實(shí)驗(yàn)組和對照組子宮平滑肌組織，，利用免疫組織化學(xué)、RT-PCR、 western bloting實(shí)驗(yàn)方法檢測Myocardin、平滑肌分化標(biāo)志基因SM22α、ERα、平滑肌增值標(biāo)志基因PCNA在SD大鼠正常子宮平滑肌和子宮肌瘤組織的表達(dá)。 3.原代細(xì)胞培養(yǎng)（SD大鼠原代子宮肌瘤細(xì)胞、正常子宮平滑肌細(xì)胞），檢測SM22-LUC、SM22mut-LUC、SM22Emut-LUC的熒光素酶活性和子宮平滑肌細(xì)胞分化與增殖marker的表達(dá)。 4.在COS-7細(xì)胞培養(yǎng)中，Co-IP檢測myocardin和ERα之間的蛋白蛋白相互作用。 結(jié)果： 1.肉眼觀察和HE染色驗(yàn)證了SD大鼠子宮肌瘤模型。 2. RT-PCR、免疫組化、western bloting檢測結(jié)果，SM22α和Myocardin的表達(dá)正常子宮平滑肌組織中比子宮肌瘤組織中的表達(dá)量高�？墒荘CNA、ERα的表達(dá)在子宮肌瘤組織中比正常子宮平滑肌組織中含量高。 3.原代細(xì)胞中LUC結(jié)果顯示myocardin驅(qū)動SM22Luciferease的活性，而ERα抑制myocardin驅(qū)動的SM22Luciferease的轉(zhuǎn)錄活性。 4. Co-IP證實(shí)ERα和myocardin之間蛋白蛋白形成復(fù)合物。 結(jié)論： 1. Myocardin通過與CArG-box結(jié)合激活下游分化基因SM22α的轉(zhuǎn)錄和蛋白表達(dá)。 2. ERα抑制myocardin對SM22α的轉(zhuǎn)錄和表達(dá)，進(jìn)而抑制子宮平滑肌細(xì)胞的分化，促進(jìn)增殖。 3. myocardin與ERα之間形成復(fù)合物；進(jìn)而可能影響Myocardin-SRF與CArG-box結(jié)合，抑制其驅(qū)動的子宮平滑肌細(xì)胞的分化作用；最終可能促進(jìn)子宮肌瘤的發(fā)生發(fā)展。
[Abstract]:Objective:. 1. To study the expression of differentiation marker gene induced by Myocardin in uterine smooth muscle cells. 2. To study the role of ER 偽 in promoting the proliferation of uterine smooth muscle cells. 3. To study the transcriptional activation and expression of differentiation marker gene induced by myocardin induced by ER 偽 in uterine smooth muscle cells. 4. Study the protein protein interaction between ER 偽 and myocardin. Methods:. 1. The model of uterine leiomyoma in SD rats was established. 2. The uterine smooth muscle tissues of the experimental group and the control group were taken after the establishment of the model of uterine leiomyoma in SD rats. The expression of Myocardinin, smooth muscle differentiation marker gene SM22 偽 er 偽 and smooth muscle value-added marker PCNA in normal uterine smooth muscle and uterine leiomyoma of SD rats was detected by immunohistochemistry RT-PCR and western bloting assay. 3. The luciferase activity of SM22-LUCU SM22mut-LUCU SM22Emut-LUC and the expression of marker in differentiation and proliferation of uterine smooth muscle cells were detected by primary cell culture and normal uterine smooth muscle cells. 4. The protein protein interaction between myocardin and ER 偽 was detected by Co-IP in COS-7 cell culture. Results:. 1. The model of uterine leiomyoma in SD rats was verified by naked eye observation and HE staining. The expression of SM22 偽 and Myocardin in normal uterine smooth muscle tissue was higher than that in uterine leiomyoma tissue, but the expression of PCNAgner 偽 in uterine leiomyoma tissue was higher than that in normal uterine smooth muscle tissue. 2. The expression of SM22 偽 and Myocardin in normal uterine smooth muscle tissue was higher than that in uterine leiomyoma tissue. 3. The results of LUC showed that SM22Luciferease was activated by myocardin, while ER 偽 inhibited the transcriptional activity of SM22Luciferease driven by myocardin. 4. Co-IP confirmed the protein formation complex between ER 偽 and myocardin. Conclusion:. 1. Myocardin activates transcription and protein expression of downstream differentiation gene SM22 偽 by binding to CArG-box. 2.ER 偽 inhibits the transcription and expression of SM22 偽 by myocardin, thus inhibits the differentiation of uterine smooth muscle cells and promotes the proliferation. 3.The formation of complex between myocardin and ER 偽 may affect the binding of Myocardin-SRF and CArG-box, inhibit the differentiation of uterine smooth muscle cells driven by Myocardin-SRF, and eventually promote the development of uterine leiomyoma.
【學(xué)位授予單位】：武漢科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R737.33

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