MRPL35蛋白在结直肠癌发生发展中作用的初步研究

发布时间：2018-03-31 21:32

本文选题：MRPL35　切入点：结直肠癌　出处：《山西医科大学》2017年硕士论文

【摘要】：背景及目标:线粒体核糖体蛋白(MRPs)于胞浆合成并经靶向输送至线粒体后,组装为线粒体核糖体,参与线粒体蛋白的翻译过程。相关的研究表明,MRPs在多种恶性肿瘤中的表达出现异常。我们前期研究结果发现结直肠癌细胞中MRPL35的表达水平上调,本文将初步分析MRPL35在结直肠癌发生发展中的作用。方法:1.收集2005-2006年的151例结直肠癌患者的组织蜡块,制作组织芯片,通过免疫组化检测MRPL35的表达;分析26对结直肠癌患者组织中MRPL35 m RNA和蛋白的表达水平;运用Kaplan Meier方法绘制生存曲线;对变量进行单因素/多因素COX回归分析,筛选独立预后因素。2.荧光定量PCR和Western blot检测结直肠癌及癌旁组织以及4种结直肠癌细胞中MRPL35 mRNA和蛋白水平的表达。慢病毒介导的RNAi敲低细胞中MRPL35的表达水平,分别采用MTT实验、克隆形成实验以及流式细胞术的方法,观察MRPL35对细胞增殖、凋亡和细胞周期的作用。结果:1.组织芯片结果显示MRPL35主要在胞浆表达,且结直肠癌组织中的表达高于癌旁组织(P0.05)。MRPL35的表达水平与淋巴结转移、远处转移、TNM分期以及病理类型呈正相关(P0.05),与病人术后生存期呈负相关(P0.05)。2.qPCR及Western blot结果显示癌组织中MRPL35的表达水平高于癌旁组织(P0.05);4种结直肠癌细胞中MRPL35表达水平均高于正常结直肠粘膜细胞FHC。3.RNAi敲低HCT116和SW480细胞的MRPL35,感染5天后,HCT116细胞的增殖能力减弱85%左右,SW480细胞的增殖能力减弱71%左右。HCT116细胞的凋亡率增加72%,SW480细胞的凋亡率增加100%。HCT116和SW480的细胞周期停滞在G2/M期。结论:MRPL35促进结直肠癌细胞增殖,其表达水平升高与患者术后生存期有密切的关系,具体分子机制有待进一步研究。
[Abstract]:Background and objective: mitochondrial ribosomal protein (MRPs) was synthesized in cytoplasm and transported to mitochondria to form mitochondrial ribosomes. Participate in the translation of mitochondrial protein. Related studies show that the expression of MRPL35 is abnormal in many kinds of malignant tumors. Our previous study found that the expression of MRPL35 in colorectal cancer cells was up-regulated. In this paper, we will analyze the role of MRPL35 in the development of colorectal cancer. Methods: 1. Collect the tissue wax of 151 patients with colorectal cancer from 2005 to 2006, make tissue microarray, and detect the expression of MRPL35 by immunohistochemistry. The expression level of MRPL35 m RNA and protein in 26 pairs of colorectal cancer tissues was analyzed, the survival curve was drawn by Kaplan Meier method, and the single factor / multivariate COX regression analysis was performed on the variables. To screen independent prognostic factors .2. fluorescent quantitative PCR and Western blot were used to detect the expression of MRPL35 mRNA and protein in colorectal cancer and adjacent tissues and four kinds of colorectal cancer cells. The expression of MRPL35 in RNAi knockout cells mediated by lentivirus was detected by MTT assay. The effects of MRPL35 on cell proliferation, apoptosis and cell cycle were observed by clone formation assay and flow cytometry. Results: 1. Tissue microarray showed that MRPL35 was mainly expressed in cytoplasm. The expression level of P0.05P. MRPL35 and lymph node metastasis in colorectal cancer tissues were higher than those in adjacent tissues. There was a positive correlation between TNM staging and pathological type of distant metastasis, and a negative correlation between MRPL35 expression and survival time after operation. The results showed that the expression of MRPL35 in cancer tissues was higher than that in adjacent tissues. The expression of MRPL35 was higher than that in the adjacent tissues of colorectal carcinoma P0.05. 2. QPCR and Western blot showed that the expression of MRPL35 was higher than that of P0. 05 in the adjacent tissues of colorectal cancer. After 5 days of infection, the proliferative ability of HCT116 cells decreased by 85% or so. The proliferative ability of SW480 cells decreased by 71%. The apoptosis rate of HCT116 cells increased by 72%. The apoptosis rate of SW480 cells increased by 100%.HCT116. And SW480 cell cycle arrest in G _ 2 / M phase. Conclusion: MRPL35 can promote the proliferation of colorectal cancer cells. The increase of expression level is closely related to the survival time after operation, and the specific molecular mechanism needs further study.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.34

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