本文選題：去勢(shì)抵抗性前列腺癌 + FKBP51�。� 參考：《天津醫(yī)科大學(xué)》2016年博士論文

【摘要】：目的:檢測(cè)在人前列腺組織及裸鼠人前列腺癌異種種植瘤中FKBP51的表達(dá)情況,人工構(gòu)建雄激素非依賴性前列腺癌細(xì)胞LNCaP-AI,并動(dòng)態(tài)檢測(cè)去勢(shì)抵抗過程中細(xì)胞內(nèi)FKBP51基因表達(dá)的變化及其在CRPC發(fā)生所起的作用,初步探究FKBP51在CRPC發(fā)生過程中上下游信號(hào)通路的改變以及長(zhǎng)鏈非編碼RNA在其中發(fā)揮的作用,探索前列腺癌由ADPC轉(zhuǎn)變?yōu)镃RPC的可能機(jī)制。方法:利用免疫組織化學(xué)技術(shù)檢測(cè)人前列腺組織及裸鼠人前列腺癌異種種植瘤中FKBP51的表達(dá);持續(xù)使用無雄激素培養(yǎng)基培養(yǎng)LNCaP細(xì)胞,人工構(gòu)建雄激素非依賴性前列腺癌細(xì)胞LNCaP-AI;通過western blot動(dòng)態(tài)檢測(cè)前列腺癌去勢(shì)抵抗過程中FKBP51基因表達(dá)的變化及其在CRPC發(fā)生所起的作用,以及其相關(guān)的NF-κB信號(hào)通路與AKT信號(hào)通路蛋白的變化;利用在線分析軟件catRAPID、Lnc RNA芯片、RNA免疫共沉淀實(shí)驗(yàn)分析前列腺癌去勢(shì)抵抗過程中長(zhǎng)鏈非編碼RNA的表達(dá)變化情況及其參與FKBP51作用信號(hào)通路改變的可能機(jī)制。結(jié)果:1.FKBP51在人前列腺癌組織標(biāo)本及裸鼠異種種植瘤標(biāo)本中均有較高表達(dá),且其表達(dá)在前列腺癌進(jìn)展為去勢(shì)抵抗時(shí)明顯升高。2.LNCa P前列腺癌細(xì)胞在無雄激素條件下培養(yǎng)1個(gè)月,細(xì)胞生長(zhǎng)速度明顯放緩且大量死亡,培養(yǎng)3個(gè)月后細(xì)胞生長(zhǎng)逐漸加快,成為雄激素非依賴的LNCaP-AI細(xì)胞亞系,并對(duì)抗雄藥物比卡魯胺及MDV3100產(chǎn)生耐藥。LNCaP-AI細(xì)胞中FKBP51表達(dá)量明顯升高,且敲除FKBP51后細(xì)胞生長(zhǎng)受到抑制。3.在去雄培養(yǎng)LNCaP-AI細(xì)胞系過程中,AR-v7表達(dá)量逐漸增高,且可不依賴雄激素調(diào)控KFBP51的表達(dá)。在LNCaP-AI細(xì)胞中FKBP51主要與IKK結(jié)合參與NF-κB信號(hào)通路的調(diào)節(jié),使caspase-3蛋白表達(dá)降低,BCL-2蛋白表達(dá)增高,從而抑制前列腺癌細(xì)胞的凋亡,而其對(duì)AKT信號(hào)通路無明顯作用。4.在列腺癌去勢(shì)抵抗過程中,多種長(zhǎng)鏈非編碼RNA的表達(dá)發(fā)生明顯變化,其中PCAT-1表達(dá)量升高,進(jìn)一步研究發(fā)現(xiàn)PCT-1可以通過與FKBP51結(jié)合,封閉FKBP51與PHLPP結(jié)合的位點(diǎn),阻礙PHLPP對(duì)AKT的降磷酸化作用,從而影響FKBP51對(duì)下游的信號(hào)通路的調(diào)節(jié)。結(jié)論:本實(shí)驗(yàn)通過構(gòu)建雄激素非依賴性前列腺癌細(xì)胞模型LNCaP-AI,證實(shí)FKBP51在去勢(shì)抵抗性前列腺癌的發(fā)生發(fā)展中發(fā)揮重要作用,其通過與長(zhǎng)鏈非編碼RNA PCAT-1共同作用,調(diào)節(jié)NF-κB及AKT信號(hào)通路,促進(jìn)前列腺癌細(xì)胞的生長(zhǎng)并抑制其凋亡,從而促進(jìn)去勢(shì)抵抗性前列腺癌的發(fā)生。本研究提示FKBP51有可能成為治療去勢(shì)抵抗性前列腺癌的一個(gè)新的藥物靶點(diǎn)。
[Abstract]:Objective: to detect the expression of FKBP51 in human prostate tissues and xenoimplants of human prostate cancer in nude mice. Androgen independent prostate cancer cell line LNCaP-AIwas constructed, and the changes of FKBP51 gene expression and its role in the pathogenesis of CRPC were dynamically detected during castration resistance. To explore the changes of upstream and downstream signaling pathways of FKBP51 in the pathogenesis of CRPC and the role of long chain non-coding RNA in the process, and to explore the possible mechanism of the transition of prostate cancer from ADPC to CRPC. Methods: immunohistochemical technique was used to detect the expression of FKBP51 in human prostate tissue and xenoimplant tumor of nude mice, androgen free medium was used to culture LNCaP cells. The androgen-independent prostate cancer cell line LNCaP-AIwas constructed, and the expression of FKBP51 gene and its role in the development of CRPC were dynamically detected by western blot during castration resistance of prostate cancer. The changes of NF- 魏 B signaling pathway and AKT signal pathway protein; The expression of long chain non-coding RNA during castration resistance of prostate cancer and its possible mechanism involved in the changes of signaling pathway of FKBP51 were analyzed by using the on-line analysis software catRIDD LNC RNA microarray and co-immunoprecipitation assay. Results 1. FKBP51 was highly expressed in both human prostate cancer tissues and xenoimplants of nude mice, and the expression of FKBP51 increased significantly when prostate cancer progressed to castration resistance. 2. LNCa P prostate cancer cells were cultured without androgen for 1 month. After 3 months of culture, the cells grew faster and became androgen independent LNCaP-AI cell sublines, and the expression of FKBP51 in the anti-androgen cells was significantly higher than that in Carous amine and MDV3100 resistant. LNCaP-AI cells. The cell growth was inhibited after knockout of FKBP51. The expression of AR-v7 in LNCaP-AI cell line was increased gradually, and the expression of KFBP51 was not regulated by androgen. In LNCaP-AI cells, FKBP51 and IKK are involved in the regulation of NF- 魏 B signaling pathway, which can decrease the expression of caspase-3 protein and increase the expression of BCL-2 protein, thus inhibit the apoptosis of prostate cancer cells, but it has no obvious effect on AKT signaling pathway. During ovariectomized resistance, the expression of long chain non-coding RNA was significantly changed, and the expression of PCAT-1 was increased. It was further found that PCT-1 could block the site of FKBP51 binding to PHLPP by binding to FKBP51. Blocking the dephosphorylation of AKT by PHLPP, thus affecting the regulation of downstream signal pathway by FKBP51. Conclusion: this study demonstrated that FKBP51 plays an important role in the development of castrated resistant prostate cancer through the establishment of androgen independent prostate cancer cell line LNCaP-AII. it works together with long chain noncoding RNA PCAT-1. Regulation of NF- 魏 B and AKT signaling pathway to promote the growth of prostate cancer cells and inhibit their apoptosis, thus promoting the development of castrated resistant prostate cancer. This study suggests that FKBP51 may be a new drug target for ovariectomized prostate cancer.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R737.25

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