ERa抑制Myocardin诱导的子宫平滑肌细胞分化

发布时间：2018-02-26 23:00

本文关键词： SD大鼠子宫肌瘤模型子宫平滑肌细胞的分化和增殖子宫肌瘤 myocardin ERα　出处：《武汉科技大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的： 1.研究Myocardin诱导的子宫平滑肌细胞分化标志基因的表达 2.研究ERα促进子宫平滑肌细胞的增值作用 3.研究ERα所抑制的myocardin诱导的子宫平滑肌细胞分化标志基因的转录激活和表达 4.研究ERα和myocardin之间的蛋白蛋白相互作用方法： 1.建立SD大鼠子宫肌瘤模型。 2. SD大鼠子宫肌瘤模型建立后分别取实验组和对照组子宫平滑肌组织，，利用免疫组织化学、RT-PCR、 western bloting实验方法检测Myocardin、平滑肌分化标志基因SM22α、ERα、平滑肌增值标志基因PCNA在SD大鼠正常子宫平滑肌和子宫肌瘤组织的表达。 3.原代细胞培养（SD大鼠原代子宫肌瘤细胞、正常子宫平滑肌细胞），检测SM22-LUC、SM22mut-LUC、SM22Emut-LUC的荧光素酶活性和子宫平滑肌细胞分化与增殖marker的表达。 4.在COS-7细胞培养中，Co-IP检测myocardin和ERα之间的蛋白蛋白相互作用。结果： 1.肉眼观察和HE染色验证了SD大鼠子宫肌瘤模型。 2. RT-PCR、免疫组化、western bloting检测结果，SM22α和Myocardin的表达正常子宫平滑肌组织中比子宫肌瘤组织中的表达量高。可是PCNA、ERα的表达在子宫肌瘤组织中比正常子宫平滑肌组织中含量高。 3.原代细胞中LUC结果显示myocardin驱动SM22Luciferease的活性，而ERα抑制myocardin驱动的SM22Luciferease的转录活性。 4. Co-IP证实ERα和myocardin之间蛋白蛋白形成复合物。结论： 1. Myocardin通过与CArG-box结合激活下游分化基因SM22α的转录和蛋白表达。 2. ERα抑制myocardin对SM22α的转录和表达，进而抑制子宫平滑肌细胞的分化，促进增殖。 3. myocardin与ERα之间形成复合物；进而可能影响Myocardin-SRF与CArG-box结合，抑制其驱动的子宫平滑肌细胞的分化作用；最终可能促进子宫肌瘤的发生发展。
[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 伪 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 伪 in uterine smooth muscle cells. 4. Study the protein protein interaction between ER 伪 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 伪 er 伪 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 伪 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 伪 and Myocardin in normal uterine smooth muscle tissue was higher than that in uterine leiomyoma tissue, but the expression of PCNAgner 伪 in uterine leiomyoma tissue was higher than that in normal uterine smooth muscle tissue. 2. The expression of SM22 伪 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 伪 inhibited the transcriptional activity of SM22Luciferease driven by myocardin. 4. Co-IP confirmed the protein formation complex between ER 伪 and myocardin. Conclusion:. 1. Myocardin activates transcription and protein expression of downstream differentiation gene SM22 伪 by binding to CArG-box. 2.ER 伪 inhibits the transcription and expression of SM22 伪 by myocardin, thus inhibits the differentiation of uterine smooth muscle cells and promotes the proliferation. 3.The formation of complex between myocardin and ER 伪 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.
【学位授予单位】：武汉科技大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.33

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