發(fā)布時(shí)間：2018-05-13 15:17

  本文選題：雄激素非依賴前列腺癌 + 上皮間質(zhì)轉(zhuǎn)化��； 參考：《上海交通大學(xué)》2014年博士論文

【摘要】：目的:抗雄性激素療法在前列腺癌臨床治療的初期效果顯著,但不能夠殺滅所有的腫瘤細(xì)胞,最終導(dǎo)致雄激素非依賴前列腺癌(AIPC)的發(fā)生,目前仍然是前列腺癌治療中的焦點(diǎn)和難點(diǎn)。殘存的、雄激素非依賴的腫瘤細(xì)胞可能具有與其他雄激素依賴的腫瘤細(xì)胞不同的生物學(xué)特性,這些細(xì)胞的形態(tài)學(xué)及其他特性目前尚不清楚。相關(guān)研究表明,上皮間質(zhì)轉(zhuǎn)化(EMT)在腫瘤的發(fā)生、發(fā)展、轉(zhuǎn)移及耐藥等多方面起著很重要的作用。本課題旨在探索和分析前列腺癌從雄激素依賴轉(zhuǎn)變成雄激素非依賴過程中EMT轉(zhuǎn)錄因子Zeb1是否及如何參與前列腺癌的雄激素非依賴的形成。方法:從細(xì)胞、組織和動(dòng)物水平利用實(shí)時(shí)定量PCR、免疫組化、免疫熒光、免疫印跡等實(shí)驗(yàn)技術(shù)分別從mRNA水平和蛋白水平來檢測(cè)Zeb1的表達(dá)情況。通過慢病毒干擾和過表達(dá)的方法通過一系列體內(nèi)體外實(shí)驗(yàn)來探討下調(diào)和上調(diào)Zeb1之后細(xì)胞的形態(tài)及功能學(xué)改變。計(jì)數(shù)數(shù)據(jù)采用χ2檢驗(yàn)方法,單因素的比較采用t檢驗(yàn)。統(tǒng)計(jì)分析和作圖采用Prism GraphPad5。P0.05被認(rèn)為具有統(tǒng)計(jì)學(xué)意義。結(jié)果:與雄激素依賴的前列腺癌細(xì)胞系(LNCa P,PC3 AR(+)和22RV1)相比,雄激素非依賴前列腺癌細(xì)胞系中(C4-2B,PC3和DU145),Zeb1的表達(dá)明顯增加,伴隨著Vimentin表達(dá)的增加和E-cadherin表達(dá)的減少。在去勢(shì)后的PTEN條件性敲除小鼠前列腺腫瘤組織中,Zeb1的表達(dá)明顯升高伴隨著上皮標(biāo)志E-cadherin和間質(zhì)標(biāo)志Vimentin的共表達(dá)。另外,與良性前列腺增生組織相比,前列腺癌組織標(biāo)本中Zeb1的表達(dá)明顯升高,而且與前列腺癌的轉(zhuǎn)移密切相關(guān)。進(jìn)一步過表達(dá)Zeb1之后,細(xì)胞對(duì)抗雄激素藥物的耐受性增加,遷移能力增強(qiáng)。體內(nèi)移植瘤實(shí)驗(yàn)發(fā)現(xiàn),去勢(shì)之后腫瘤仍然能夠繼續(xù)生長。而干擾Zeb1的表達(dá)后,細(xì)胞對(duì)抗雄激素藥物的敏感性增加,遷移能力下降。有趣的是,Zeb1的表達(dá)伴隨著一些多能干細(xì)胞標(biāo)志物如Sox2的變化,以及細(xì)胞克隆形成能力和成球能力的改變。同時(shí),對(duì)小鼠進(jìn)行去勢(shì)處理之后,小鼠前列腺組織中Zeb1與Sox2的表達(dá)均出現(xiàn)升高。進(jìn)一步免疫沉淀發(fā)現(xiàn)Zeb1能夠特異性的與Sox2蛋白相結(jié)合。結(jié)論:EMT轉(zhuǎn)錄調(diào)控因子Zeb1可能通過誘導(dǎo)干細(xì)胞樣特性促進(jìn)前列腺癌雄激素非依賴的形成和轉(zhuǎn)移。調(diào)控Zeb1抑制上皮間質(zhì)轉(zhuǎn)化過程,或許可以為前列腺癌的臨床治療提供潛在的可能分子靶標(biāo)。
[Abstract]:Objective: antiandrogen therapy is effective in the early stage of clinical treatment of prostate cancer, but it can not kill all tumor cells, leading to the development of androgen independent prostate cancer (AICC). At present, prostate cancer treatment is still the focus and difficulty. The remaining androgen independent tumor cells may have different biological characteristics from those of other androgen dependent tumor cells, the morphological and other characteristics of which are unclear. Related studies have shown that EMTs play an important role in carcinogenesis, development, metastasis and drug resistance. The purpose of this study was to explore whether and how EMT transcription factor Zeb1 is involved in the formation of androgen independence in prostate cancer from androgen dependent to androgen independent. Methods: the expression of Zeb1 was detected by real-time quantitative PCR, immunohistochemistry, immunofluorescence and Western blotting at cell, tissue and animal levels, respectively. By means of lentivirus interference and overexpression, a series of experiments in vitro and in vivo were carried out to investigate the morphological and functional changes of Zeb1 after down-regulation and up-regulation. 蠂 2 test was used for counting data and t test was used for single factor comparison. The use of Prism GraphPad5.P0.05 in statistical analysis and mapping is considered statistically significant. Results: compared with androgen-dependent prostate cancer cell lines (LNCA PnPC3AR3 () and 22RV1), androgen independent prostate cancer cell lines increased the expression of C4-2BmPC3 and DU145ZEb1, accompanied by the increase of Vimentin expression and the decrease of E-cadherin expression. In castrated PTEN conditioned knockout mice, the expression of Zeb1 was significantly increased with the coexpression of epithelial marker E-cadherin and interstitial marker Vimentin. In addition, the expression of Zeb1 in prostate cancer tissues was significantly higher than that in benign prostatic hyperplasia tissues, and was closely related to the metastasis of prostate cancer. After further overexpression of Zeb1, cell resistance to androgen drugs increased and migration ability increased. In vivo transplanted tumor experiments found that the tumor can continue to grow after castration. After interfering with the expression of Zeb1, the sensitivity of the cells to androgen drugs was increased, and the migration ability was decreased. Interestingly, the expression of Zeb1 was accompanied by changes in some pluripotent stem cell markers such as Sox2, as well as the ability of cell clone formation and spherulation. At the same time, the expression of Zeb1 and Sox2 in the prostate tissues of mice increased after castration. Further immunoprecipitation revealed that Zeb1 could specifically bind to Sox2 protein. Conclusion Zeb1 may promote androgen independent development and metastasis of prostate cancer by inducing stem cell-like characteristics. Regulating the inhibition of epithelial mesenchymal transformation by Zeb1 may provide a potential molecular target for the clinical treatment of prostate cancer.
【學(xué)位授予單位】：上海交通大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R737.25

【共引文獻(xiàn)】

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1 鐘寶元;劉艷秀;;腫瘤中STAT3、P-STAT3的表達(dá)及與EMT關(guān)系的研究進(jìn)展[J];贛南醫(yī)學(xué)院學(xué)報(bào);2013年04期

2 張建蓉;佘強(qiáng);;上皮-間質(zhì)轉(zhuǎn)化在心臟發(fā)育和損傷中的作用[J];國際心血管病雜志;2013年05期

3 張磊;吳繼鋒;孫雪竹;孫景洲;;胃癌組織中Twist、STAT3和E-cad的表達(dá)及其臨床意義[J];蚌埠醫(yī)學(xué)院學(xué)報(bào);2013年10期

4 彭璇;;ZEB-1在胃癌組織中的表達(dá)及臨床意義[J];湖北民族學(xué)院學(xué)報(bào)(醫(yī)學(xué)版);2013年03期

5 流小舟;黎承軍;周光新;趙建寧;;核轉(zhuǎn)錄因子Snail與骨肉瘤關(guān)系的研究進(jìn)展[J];東南國防醫(yī)藥;2014年01期

6 Xiang-Ming Ding;;MicroRNAs: regulators of cancer metastasis and epithelialmesenchymal transition(EMT)[J];Chinese Journal of Cancer;2014年03期

7 李f3穎;姜樺;;ZEB家族在上皮性卵巢癌上皮-間質(zhì)轉(zhuǎn)化中的作用[J];國際婦產(chǎn)科學(xué)雜志;2014年01期

8 王一;黎燕;沈倍奮;陳國江;;炎癥:腫瘤的推進(jìn)器[J];國際藥學(xué)研究雜志;2014年01期

9 陳杰;郭維華;田衛(wèi)東;;牙囊和上皮根鞘細(xì)胞成無細(xì)胞牙骨質(zhì)的機(jī)制[J];國際口腔醫(yī)學(xué)雜志;2014年03期

10 章俊;邱敏姿;馬亞瓊;卜陽;楊蕾;湯s，

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