發(fā)布時(shí)間：2018-06-21 02:00

  本文選題：HCC + Rap1b��； 參考：《重慶醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的研究Rap1b在肝細(xì)胞肝癌發(fā)生發(fā)展過程中的作用及其相關(guān)機(jī)制探討。方法1.采用免疫組織化學(xué)染色和q RT-PCR技術(shù),檢測(cè)Rap1b在人肝癌組織和癌旁組織中表達(dá)情況。采用q RT-PCR和western blot方法,檢測(cè)肝癌細(xì)胞中Rap1b的表達(dá)情況。2.運(yùn)用基因重組技術(shù)構(gòu)建Rap1b過表達(dá)質(zhì)粒和合成Rap1b的干擾片段。轉(zhuǎn)染入肝癌細(xì)胞系中,采用MTS和克隆形成實(shí)驗(yàn),分析Rap1b對(duì)HCC增殖功能的影響。采用劃痕實(shí)驗(yàn)和transwell實(shí)驗(yàn),檢測(cè)Rap1b對(duì)HCC侵襲和遷移功能的影響。3.運(yùn)用分子生物學(xué)技術(shù)及查閱相關(guān)文獻(xiàn),選取影響腫瘤細(xì)胞侵襲和遷移功能的分子。運(yùn)用q RT-PCR方法,檢測(cè)Rap1b對(duì)相關(guān)分子表達(dá)量的影響。采用蛋白質(zhì)印跡技術(shù),測(cè)定預(yù)測(cè)靶基因在蛋白水平的變化。再選用雙熒光素酶報(bào)告基因?qū)嶒?yàn),測(cè)定Rap1b與靶基因啟動(dòng)子之間的關(guān)系。4.在裸鼠體內(nèi),成瘤實(shí)驗(yàn)測(cè)定用于分析腫瘤形成能力。腫瘤遷移試驗(yàn)用于分析腫瘤遷移能力。結(jié)果1.Rap1b在人肝癌組織和肝癌細(xì)胞中表達(dá)量升高。2.Rap1b在體外促進(jìn)肝癌細(xì)胞的增殖與侵襲遷移功能。3.Rap1b通過增強(qiáng)其啟動(dòng)子活性從而上調(diào)Twist1表達(dá)量。4.Rap1b在體內(nèi)能夠促進(jìn)腫瘤的形成與遷移侵襲能力。結(jié)論Rap1b通過靶向Twist1從而影響肝癌細(xì)胞的發(fā)生發(fā)展。
[Abstract]:Objective to study the role of Rap1b in the development of hepatocellular carcinoma (HCC). Method 1. The expression of Rap1b in human hepatocellular carcinoma and paracancerous tissues was detected by immunohistochemical staining and Q RT-PCR. The expression of Rap1b in hepatocellular carcinoma cells was detected by Q RT-PCR and western blot. Rap1b overexpression plasmid was constructed by gene recombination technique and the interference fragment of Rap1b was synthesized. The effect of Rap1b on the proliferation of HCC was analyzed by MTS and clone formation assay. The effect of Rap1b on invasion and migration of HCC was detected by scratch test and transwell test. Molecular biological techniques and literature were used to select the molecules that affect the invasion and migration of tumor cells. The effect of Rap1b on the expression of related molecules was detected by Q RT-PCR. Western blotting technique was used to detect and predict the changes of target genes at protein level. The relationship between Rap1b and target gene promoter was determined by double luciferase reporter gene experiment. In nude mice, tumorigenesis assay was used to analyze tumor formation ability. Tumor migration test is used to analyze the ability of tumor metastasis. 2. Rap1b promoted the proliferation, invasion and migration of hepatoma cells in vitro. 3. Rap1b upregulated the expression of Twist1 by enhancing its promoter activity. 4. Rap1b could promote tumor growth in vivo. Ability to form and migrate invasion. Conclusion Rap1b can affect the development of hepatoma cells by targeting Twist1.
【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
【分類號(hào)】：R735.7

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

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本文編號(hào)：2046675