本文選題：雌激素　切入點(diǎn)：雌激素受體α36　出處：《重慶醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:本實(shí)驗(yàn)旨在研究雌激素受體α36(Estrogen receptor alpha-36,ERα36)在介導(dǎo)雌激素促進(jìn)甲狀腺乳頭狀癌(Papillary thyroid carcinoma,PTC)增殖和侵襲轉(zhuǎn)移的分子機(jī)制及其相關(guān)信號通路。方法:用免疫組織化學(xué)SP法對PTC組織進(jìn)行ERα36蛋白檢測,分析其表達(dá)情況與臨床病理特征的相關(guān)性,同時檢測PTC組織中EGFR和HER2的表達(dá),分析三者表達(dá)之間的相關(guān)性。Western blot方法檢測PTC細(xì)胞株K-1和BCPAP中ERα36的表達(dá)情況;Western blot檢測E2對K-1和BCPAP細(xì)胞ERK1/2、AKT磷酸化水平和NF-κB核轉(zhuǎn)位的影響,以及ERα36-si RNA對其的抑制作用;Western blot檢測NF-κB下游靶基因Cyclin Dl、Survivin和MMP2的蛋白表達(dá)情況,以及ERα36-siRNA的抑制作用;梯度濃度的E2處理BCPAP細(xì)胞,MTT法檢測其細(xì)胞增殖情況;Transwell小室實(shí)驗(yàn)檢測BCPAP細(xì)胞侵襲遷移能力。結(jié)果:在218例PTC組織中,ERα36、EGFR和HER2的陽性表達(dá)率分別為112(51.4%)、132(60.6%)和135(61.9%),明顯高于正常組織和結(jié)節(jié)性增生(P0.001)。ERα36、EGFR和HER2的表達(dá)與TNM分期和頸部淋巴結(jié)轉(zhuǎn)移顯著相關(guān)(P0.001)。此外,ERα36與EGFR的表達(dá)呈正相關(guān)(rs=0.285,P0.001),ERα36與HER2的表達(dá)呈正相關(guān)(rs=0.352,P0.001)。兩種蛋白(ERα36/EGFR、ERα36/HER2和EGFR/HER2)聯(lián)合表達(dá)和僅一種蛋白表達(dá)相比,與頸部淋巴結(jié)轉(zhuǎn)移和TNM分期更具相關(guān)性(P0.001)。ERα36、EGFR和HER2三種蛋白聯(lián)合表達(dá)較僅一種或兩種蛋白表達(dá)與頸部淋巴結(jié)轉(zhuǎn)移和TNM分期更具有相關(guān)性(P0.001)。PTC細(xì)胞株K-1和BCPAP均表達(dá)ERα36;E2處理細(xì)胞的最佳條件是10-8mol/L持續(xù)24h。E2能促進(jìn)BCPAP和K-1細(xì)胞ERK1/2和AKT蛋白磷酸化水平增高,且在15min時BCPAP細(xì)胞最明顯,10min時K-1細(xì)胞最明顯,ERα36-si RNA能抑制E2的誘導(dǎo)效果;隨著E2處理時間的增加,核NF-κB p65表達(dá)水平逐漸升高,細(xì)胞質(zhì)NF-κB p65逐漸降低,即E2促進(jìn)BCPAP和K-1細(xì)胞發(fā)生NF-κB核轉(zhuǎn)位,ERα36-si RNA抑制E2誘導(dǎo)的核轉(zhuǎn)位;E2促進(jìn)Cyclin Dl、Survivin和MMP2的表達(dá),ERα36-siRNA抑制該作用;E2促進(jìn)BAPAP細(xì)胞增殖,ERα36-siRNA則抑制E2的誘導(dǎo)效果;ERα36-siRNA抑制E2誘導(dǎo)的BAPAP細(xì)胞侵襲遷移能力。結(jié)論:ERα36、EGFR和HER2在PTC組織中的表達(dá)與TNM分期及頸部淋巴結(jié)轉(zhuǎn)移呈顯著正相關(guān)。ERα36通過ERK/AKT-NF-κB通路介導(dǎo)雌激素促進(jìn)PTC細(xì)胞增殖和侵襲轉(zhuǎn)移。
[Abstract]:Aim: to investigate the molecular mechanism of estrogen receptor 偽 36(Estrogen receptor alpha-36 (ER 偽 36) in promoting the proliferation, invasion and metastasis of papillary thyroid carcinoma of thyroid (Papillary thyroid carcinoma-PTC36).Methods: immunohistochemical SP method was used to detect ER 偽 36 protein in PTC tissues. The correlation between the expression of ER 偽 36 protein and clinicopathological features was analyzed. The expression of EGFR and HER2 in PTC tissues was also detected.Western blot was used to detect the expression of ER 偽 36 in PTC cell line K-1 and BCPAP. Western blot was used to detect the effect of E2 on phosphorylation level and NF- 魏 B nuclear translocation in K-1 and BCPAP cells.The inhibitory effect of ER 偽 36-si RNA on the expression of survivin and MMP2, as well as the inhibitory effect of ER 偽 36-siRNA on the expression of NF- 魏 B downstream target gene Cyclin Dlnsurvivin and MMP2 were detected by Western blot.The proliferation of BCPAP cells treated with E _ 2 at gradient concentration was assayed by MTT assay. Transwell chamber assay was used to detect the invasion and migration ability of BCPAP cells.Results: the positive expression rates of ER 偽 36-EGFR and HER2 in 218 cases of PTC were 11251.4 and 1352o60.6, respectively. The positive rates of ER 偽 36-EGFR and HER2 were significantly higher than those of normal tissues and nodular hyperplasia (P0.001T, ER 偽 36-EGFR and HER2), and the expression of ER 偽 36-EGFR and HER2 were significantly correlated with TNM stage and cervical lymph node metastasis.In addition, the expression of ER 偽 36 was positively correlated with the expression of EGFR. The expression of ER 偽 36 was positively correlated with the expression of HER2.Two proteins, ER 偽 36 / EGFRN ER 偽 36/HER2 and EGFR / HER2, were expressed in comparison with only one protein.There was a significant correlation between the expression of P0.001T, ER 偽 36G, EGFR and HER2 protein and the cervical lymph node metastasis and TNM stage. The expression of ER 偽 36C E2 in P0.001PTC cell line K-1 and BCPAP was more correlated with cervical lymph node metastasis and TNM stage.The best condition for cells was that 10-8mol/L continuous 24h.E2 could increase the phosphorylation of ERK1/2 and AKT protein in BCPAP and K-1 cells.The expression of nuclear NF- 魏 B p65 increased and the cytoplasm NF- 魏 B p65 decreased with the increase of E2 treatment time, and the effect of ER 偽 36-si RNA was most obvious in K-1 cells at 10 minutes after 15min treatment, and the expression level of NF- 魏 B p65 increased gradually with the increase of E2 treatment time, and the expression of NF- 魏 B p65 in cytoplasm decreased gradually with the increase of E2 treatment time.Conclusion the expression of EGFR and HER2 in PTC tissues is positively correlated with TNM stage and cervical lymph node metastasis. ER 偽 36 mediates estrogen mediated by ERK / AKT-NF- 魏 B pathway to promote the proliferation and invasion and metastasis of PTC cells.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R736.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 趙樂;唐萃;楊磊;莫小梅;朱平;劉智敏;;Caveolin-1、GPR30和Vimentin在甲狀腺乳頭狀癌組織中的表達(dá)及意義[J];中國免疫學(xué)雜志;2015年11期

2 廉馨;史丹;李洪艷;;新型雌激素受體ER-α36與乳腺癌[J];中國生物化學(xué)與分子生物學(xué)報(bào);2015年01期

3 劉旺根;張欽憲;王云龍;孫新城;王國強(qiáng);李恒思;李玉林;;ER-α36過表達(dá)促進(jìn)人乳腺癌MCF-7細(xì)胞侵襲轉(zhuǎn)移能力[J];基礎(chǔ)醫(yī)學(xué)與臨床;2013年06期

4 鄒豐;王緒明;劉麗江;;ER-α36和miR-143介導(dǎo)胃癌細(xì)胞的侵襲[J];中國病理生理雜志;2012年12期

5 彭瑤;連增林;;新型雌激素受體ER-α36的生物學(xué)效應(yīng)及其相關(guān)疾病的研究現(xiàn)狀[J];中國醫(yī)藥生物技術(shù);2012年01期

6 趙坤;黃俊星;;EGFR、Her-2在食管鱗癌中表達(dá)的研究進(jìn)展[J];現(xiàn)代腫瘤醫(yī)學(xué);2011年08期

7 譚艷華;王學(xué)軍;;甲狀腺乳頭狀癌中細(xì)胞周期素E的表達(dá)及臨床意義[J];中國地方病防治雜志;2010年05期

8 陳竟文;宋陸軍;;甲狀腺癌的流行病學(xué)新特點(diǎn)[J];中國臨床醫(yī)學(xué);2009年05期

9 封爽;王洋;王曦;王兆一;崔玉影;劉晶;趙春暉;金梅;鄒偉;;Caveolin-1基因沉默促進(jìn)乳腺上皮細(xì)胞ERα36介導(dǎo)的PI3K/AKT信號通路的激活[J];中國科學(xué)(C輯:生命科學(xué));2009年09期

10 王s，

本文編號：1720256