Rap1b上调Twist1促进肝癌细胞的侵袭迁移能力

发布时间：2018-06-21 02:00

  本文选题：HCC + Rap1b　； 参考：《重庆医科大学》2017年硕士论文

【摘要】：目的研究Rap1b在肝细胞肝癌发生发展过程中的作用及其相关机制探讨。方法1.采用免疫组织化学染色和q RT-PCR技术,检测Rap1b在人肝癌组织和癌旁组织中表达情况。采用q RT-PCR和western blot方法,检测肝癌细胞中Rap1b的表达情况。2.运用基因重组技术构建Rap1b过表达质粒和合成Rap1b的干扰片段。转染入肝癌细胞系中,采用MTS和克隆形成实验,分析Rap1b对HCC增殖功能的影响。采用划痕实验和transwell实验,检测Rap1b对HCC侵袭和迁移功能的影响。3.运用分子生物学技术及查阅相关文献,选取影响肿瘤细胞侵袭和迁移功能的分子。运用q RT-PCR方法,检测Rap1b对相关分子表达量的影响。采用蛋白质印迹技术,测定预测靶基因在蛋白水平的变化。再选用双荧光素酶报告基因实验,测定Rap1b与靶基因启动子之间的关系。4.在裸鼠体内,成瘤实验测定用于分析肿瘤形成能力。肿瘤迁移试验用于分析肿瘤迁移能力。结果1.Rap1b在人肝癌组织和肝癌细胞中表达量升高。2.Rap1b在体外促进肝癌细胞的增殖与侵袭迁移功能。3.Rap1b通过增强其启动子活性从而上调Twist1表达量。4.Rap1b在体内能够促进肿瘤的形成与迁移侵袭能力。结论Rap1b通过靶向Twist1从而影响肝癌细胞的发生发展。
[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.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.7

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相关期刊论文 前1条

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