本文選題：NF-κB + IKKα　； 參考：《第二軍醫(yī)大學(xué)》2006年博士論文

【摘要】：近年研究表明,乳腺腫瘤細(xì)胞中,IκB激酶-α(IκB kinase-α,IKKα)對(duì)激活NF-κB乃至其后的病變發(fā)揮著極其重要的作用。腫瘤壞死因子家族成員receptor activator of NF-κB ligand(RANKL)與其受體(RANK)結(jié)合,激活I(lǐng)KKα蛋白,釋放NF-κB蛋白轉(zhuǎn)位進(jìn)入細(xì)胞核。在核內(nèi)促使CyclinD 1的表達(dá),若該表達(dá)持續(xù)進(jìn)行將會(huì)導(dǎo)致乳腺上皮細(xì)胞的增生、癌變。該通路是經(jīng)典的TNFR-IKKβ-NF-κB及LT-IKKα-NF-κB通路之外的另一重要信號(hào)轉(zhuǎn)導(dǎo)途徑,稱之為RANKL-IKKα-NF-κB通路。它的異常表達(dá)是導(dǎo)致乳腺腫瘤形成的早期事件之一。本課題針對(duì)NF-κB信號(hào)轉(zhuǎn)導(dǎo)通路中的重要樞紐因子IKKα,通過(guò)應(yīng)用慢病毒載體(lentiviral vector)介導(dǎo)的RNAi效應(yīng),剔降(knockdown)乳腺腫瘤細(xì)胞MCF-7的IKKαm RNA,使其表達(dá)量下降,從而降低下游NF-κB蛋白的過(guò)度激活,以至恢復(fù)到正常水平,避免乳腺腫瘤的發(fā)生發(fā)展。 此外,采用慢病毒載體感染受精卵方法,取代傳統(tǒng)顯微注射,建立了表達(dá)IKKα基因剔降的轉(zhuǎn)基因小鼠模型,目前已繁殖至F2代。通過(guò)實(shí)時(shí)定量PCR技術(shù)驗(yàn)證,轉(zhuǎn)基因小鼠血液中IKKα m RNA水平,明顯低于野生型小鼠。該模型的建立為在體內(nèi)開展IKKα基因的生物學(xué)功能及其與腫瘤發(fā)生關(guān)系的研究創(chuàng)造良好條件。上述研究工作在國(guó)內(nèi)外尚未見文獻(xiàn)報(bào)道。 課題研究的內(nèi)容包括以下四個(gè)主要部分: 1.pU6-IKKα-knockdown表達(dá)載體構(gòu)建。 2.瞬時(shí)轉(zhuǎn)染pU6-IKKα-knockdown表達(dá)載體剔降MCF-7細(xì)胞中IKKα的m RNA的研究。 3.IKKα-knockdown慢病毒載體的構(gòu)建、穩(wěn)定轉(zhuǎn)染及用于乳腺腫瘤基因治療的初步觀察。
[Abstract]:Recent studies have shown that I 魏 B kinase- 偽, I 魏 B kinase- 偽 and IKK 偽 play an extremely important role in the activation of NF- 魏 B and subsequent lesions in breast tumor cells. Tumor necrosis factor family member receptor activator of NF- 魏 B ligand RANKL binds to its receptor RANKL, activates IKK 偽 protein and releases NF- 魏 B protein translocation into the nucleus. The expression of CyclinD 1 is promoted in the nucleus, which, if sustained, will lead to proliferation and canceration of breast epithelial cells. This pathway is another important signal transduction pathway in addition to the classical TNFR-IKK 尾 -NF- 魏 B and LT-IKK 偽 -NF- 魏 B pathways, known as the RANKL-IKK 偽 -NF- 魏 B pathway. Its abnormal expression is one of the early events leading to breast tumor formation. In this study, IKK 偽, an important pivotal factor in NF- 魏 B signal transduction pathway, was down-regulated by the RNAi effect mediated by lentiviral vector, and the expression of IKK 偽 m in breast cancer cells was reduced, thus reducing the overexpression of NF- 魏 B protein downstream. Even return to the normal level, to avoid the occurrence and development of breast tumors. In addition, a transgenic mouse model expressing IKK 偽 gene was established by using lentivirus vector to infect fertilized eggs instead of traditional microinjection, which has been propagated to F2 generation. The level of IKK 偽 m RNA in blood of transgenic mice was significantly lower than that of wild type mice. The establishment of the model provides favorable conditions for the study of the biological function of IKK 偽 gene and its relationship with oncogenesis in vivo. The above research work has not been reported in the literature at home and abroad. The content of the research includes the following four main parts: 1.pU6-IKK 偽 -knockdown expression vector was constructed. 2. Transient transfection of pU6-IKK 偽 -knockdown expression vector to degrade m RNA of IKK 偽 in MCF-7 cells. Construction of 3.IKK 偽 -knockdown lentivirus vector, stable transfection and preliminary observation of gene therapy for breast tumor.
【學(xué)位授予單位】：第二軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2006
【分類號(hào)】：R346;R-332

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 趙延森;黃牛和水牛INHA基因RNAi慢病毒載體的構(gòu)建與鑒定及功能研究[D];華中農(nóng)業(yè)大學(xué);2011年

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本文編號(hào)：1780120