ERα46在甲状腺乳头状癌中的作用及机制研究

发布时间：2018-02-27 11:39

本文关键词： ERα46 甲状腺乳头状癌增殖凋亡　出处：《重庆医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:探索雌激素受体ERα46在甲状腺乳头状癌中的作用及分子机制。方法:Western blot检测ERα46/ERα66在甲状腺乳头状癌BCPAP细胞及正常甲状腺细胞Nthy-ori3-1中的表达。构建ERα46真核表达载体转染至BCPAP细胞,Western blot检测ERα46/ERα66表达。提取转染细胞总RNA,反转录并用实时定量PCR(RT-PCR)检测细胞中c-fos和cyclin D1 mRNA的表达情况。流式细胞技术检测ERα46过表达的细胞周期分布及细胞凋亡。细胞计数法检测ERα46过表达后BCPAP细胞的增殖情况。RT-PCR检测miRNA21、PDCD4和PTEN的表达,Western blot检测PDCD4和PTEN蛋白的表达。结果:ERα46/ERα66比率在甲状腺乳头状癌BCPAP细胞中较甲状腺正常细胞下调(P㩳0.05)。ERα46过表达后ERα46/ERα66比率在BCPAP细胞中上调,抑制雌激素诱导的BCPAP细胞中c-fos和cyclin D1的mRNA表达(P㩳0.05),抑制了雌激素诱导的BCPAP细胞周期移行(P㩳0.05),抑制雌激素诱导的BCPAP细胞增殖(P㩳0.05)。ERα46过表达促进雌激素诱导的甲状腺乳头状癌细胞凋亡。ERα46过表达细胞中miRNA21的表达明显降低(P㩳0.05)且雌激素诱导了miRNA21表达水平的进一步降低(P㩳0.05)。ERα46过表达细胞中PDCD4和PTEN的mRNA和蛋白表达水平明显升高(P㩳0.05)且雌激素进一步诱导PDCD4和PTEN的表达水平升高(P㩳0.05)。结论:ERα46过表达上调ERα46/ERα66比率抑制甲状腺乳头状癌细胞增殖,促进细胞凋亡。
[Abstract]:Objective: to investigate the role and molecular mechanism of estrogen receptor ER 伪 46 in papillary thyroid carcinoma. Methods: the expression of ER 伪 46 / ER 伪 66 in Nthy-ori3-1 of BCPAP cells and normal thyroid cells of papillary thyroid carcinoma was detected by Western blot. A eukaryotic expression vector of ER 伪 46 was constructed. The expression of ER 伪 46 / ER 伪 66 was detected by Western blot after transfection into BCPAP cells. The expression of c-fos and cyclin D1 mRNA in transfected cells was detected by reverse transcription and real-time quantitative PCRRT PCR. Flow cytometry was used to detect the cell cycle distribution and cell cycle distribution of ER 伪 46 overexpression. Apoptosis. Cell count method was used to detect the proliferation of BCPAP cells after overexpression of ER 伪 46. RT-PCR was used to detect the expression of miRNA21, PDCD4 and PTEN. Western blot was used to detect the expression of PDCD4 and PTEN protein. Results the ratio of ER 伪 46 / ER 伪 66 in BCPAP cells of papillary thyroid carcinoma was lower than that of normal thyroid cells. The ratio of ER 伪 46 / ER 伪 66 was up-regulated in BCPAP cells after overexpression of ER 伪 46, which inhibited the mRNA expression of c-fos and cyclin D1 in BCPAP cells induced by estrogen. 0. 05%, inhibiting estrogen induced cell cycle migration of BCPAP cells. Inhibition of BCPAP cell proliferation induced by estrogen? The overexpression of ER 伪 46 can promote the expression of miRNA21 in estrogen induced apoptosis of thyroid papillary carcinoma cells. And estrogen induced a further decrease in the expression of miRNA21. The expression level of mRNA and protein of PDCD4 and PTEN in the overexpression cells of 0.05 and ER 伪 46 was significantly increased. And estrogen further induced the expression of PDCD4 and PTEN increased. Conclusion the overexpression of ER 伪 46 up-regulates the ratio of ER 伪 46 / ER 伪 66 to inhibit the proliferation of thyroid papillary carcinoma cells and promote apoptosis.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R736.1

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