本文關(guān)鍵詞： ERα46 甲狀腺乳頭狀癌 增殖 凋亡　出處：《重慶醫(yī)科大學(xué)》2017年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：目的:探索雌激素受體ERα46在甲狀腺乳頭狀癌中的作用及分子機(jī)制。方法:Western blot檢測(cè)ERα46/ERα66在甲狀腺乳頭狀癌BCPAP細(xì)胞及正常甲狀腺細(xì)胞Nthy-ori3-1中的表達(dá)。構(gòu)建ERα46真核表達(dá)載體轉(zhuǎn)染至BCPAP細(xì)胞,Western blot檢測(cè)ERα46/ERα66表達(dá)。提取轉(zhuǎn)染細(xì)胞總RNA,反轉(zhuǎn)錄并用實(shí)時(shí)定量PCR(RT-PCR)檢測(cè)細(xì)胞中c-fos和cyclin D1 mRNA的表達(dá)情況。流式細(xì)胞技術(shù)檢測(cè)ERα46過(guò)表達(dá)的細(xì)胞周期分布及細(xì)胞凋亡。細(xì)胞計(jì)數(shù)法檢測(cè)ERα46過(guò)表達(dá)后BCPAP細(xì)胞的增殖情況。RT-PCR檢測(cè)miRNA21、PDCD4和PTEN的表達(dá),Western blot檢測(cè)PDCD4和PTEN蛋白的表達(dá)。結(jié)果:ERα46/ERα66比率在甲狀腺乳頭狀癌BCPAP細(xì)胞中較甲狀腺正常細(xì)胞下調(diào)(P㩳0.05)。ERα46過(guò)表達(dá)后ERα46/ERα66比率在BCPAP細(xì)胞中上調(diào),抑制雌激素誘導(dǎo)的BCPAP細(xì)胞中c-fos和cyclin D1的mRNA表達(dá)(P㩳0.05),抑制了雌激素誘導(dǎo)的BCPAP細(xì)胞周期移行(P㩳0.05),抑制雌激素誘導(dǎo)的BCPAP細(xì)胞增殖(P㩳0.05)。ERα46過(guò)表達(dá)促進(jìn)雌激素誘導(dǎo)的甲狀腺乳頭狀癌細(xì)胞凋亡。ERα46過(guò)表達(dá)細(xì)胞中miRNA21的表達(dá)明顯降低(P㩳0.05)且雌激素誘導(dǎo)了miRNA21表達(dá)水平的進(jìn)一步降低(P㩳0.05)。ERα46過(guò)表達(dá)細(xì)胞中PDCD4和PTEN的mRNA和蛋白表達(dá)水平明顯升高(P㩳0.05)且雌激素進(jìn)一步誘導(dǎo)PDCD4和PTEN的表達(dá)水平升高(P㩳0.05)。結(jié)論:ERα46過(guò)表達(dá)上調(diào)ERα46/ERα66比率抑制甲狀腺乳頭狀癌細(xì)胞增殖,促進(jìn)細(xì)胞凋亡。
[Abstract]:Objective: to investigate the role and molecular mechanism of estrogen receptor ER 偽 46 in papillary thyroid carcinoma. Methods: the expression of ER 偽 46 / ER 偽 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 偽 46 was constructed. The expression of ER 偽 46 / ER 偽 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 偽 46 overexpression. Apoptosis. Cell count method was used to detect the proliferation of BCPAP cells after overexpression of ER 偽 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 偽 46 / ER 偽 66 in BCPAP cells of papillary thyroid carcinoma was lower than that of normal thyroid cells. The ratio of ER 偽 46 / ER 偽 66 was up-regulated in BCPAP cells after overexpression of ER 偽 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 偽 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 偽 46 was significantly increased. And estrogen further induced the expression of PDCD4 and PTEN increased. Conclusion the overexpression of ER 偽 46 up-regulates the ratio of ER 偽 46 / ER 偽 66 to inhibit the proliferation of thyroid papillary carcinoma cells and promote apoptosis.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R736.1

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相關(guān)期刊論文 前1條

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本文編號(hào)：1542516