鼻咽癌转移的线粒体蛋白质组研究

发布时间：2018-06-07 14:51

本文选题：鼻咽癌 + 肿瘤转移　；参考：《中南大学》2013年博士论文

【摘要】：目的：分析和鉴定鼻咽癌转移中线粒体蛋白质组的改变,筛选与鼻咽癌转移相关的线粒体蛋白质,为进一步揭示它们参与鼻咽癌转移的分子机制和找寻与鼻咽癌转移相关的特异性分子标志物提供基础。方法：分别从鼻咽癌5-8F和6-10B细胞中分离线粒体,鉴定其纯度,提取线粒体蛋白质。经2D-DIGE技术找出两种细胞的线粒体差异蛋白质,进行MALDI-TOF和ESI-Q-TOF质谱分析,查询数据库鉴定。同时采用生物信息学方法对线粒体差异蛋白质进行GO功能富集分析和蛋白质相互作用网络构建。并对鼻咽癌5-8F细胞中表达上调的部分线粒体差异蛋白质进行siRNA瞬时干扰,Transwell迁移实验观察细胞运动能力的变化。结果：包括PRDX3和SOD2在内的16种线粒体差异蛋白质得到鉴定。在鼻咽癌5-8F细胞中抑制PRDX3的表达能够提高细胞的运动能力。GO功能富集分析显示生物学过程主要表现在细胞对活性氧应答反应、过氧化氢代谢过程、线粒体膜电势调控、细胞氧化还原动态平衡和氧化还原作用等5个方面；分子功能主要表现在氧化还原酶活性、半胱氨酸蛋白酶抑制剂活性、过氧化物酶活性、质膜通道蛋白活性和抗氧化剂活性等5个方面。经过数据挖掘,构建了一个线粒体差异蛋白质相互作用的网络。结合肿瘤-间质共演变假说的观点进行分析,初步揭示了PRDX3、PRDX6、SOD2、ECH1、SERPINB5、COX5A、 PDIA5、EIF5A、IDH3B和PSMC4等10种线粒体差异蛋白质参与鼻咽癌转移的可能分子机制,为氧化应激直接促进肿瘤转移的观点提供了实验证据。结论：共鉴定了16种线粒体差异表达蛋白质,初步揭示了其中10种参与肿瘤转移的可能分子机制。在鼻咽癌5-8F细胞中抑制PRDX3的表达能够提高细胞的运动能力。它们有可能成为鼻咽癌转移的分子标志,在线粒体氧化应激诱导的鼻咽癌转移过程中发挥重要作用。
[Abstract]:Objective: to analyze and identify the changes of mitochondrial proteome in metastasis of nasopharyngeal carcinoma (NPC), and to screen the mitochondrial proteins associated with metastasis of nasopharyngeal carcinoma (NPC). It provides a basis for further revealing the molecular mechanism of their involvement in the metastasis of nasopharyngeal carcinoma and finding specific molecular markers related to metastasis of nasopharyngeal carcinoma. Methods: mitochondria were isolated from 5-8F and 6-10B cells of nasopharyngeal carcinoma, their purity was identified and mitochondrial protein was extracted. Mitochondrial differential proteins were identified by 2D-DIGE technique and analyzed by MALDI-TOF and ESI-Q-TOF mass spectrometry. At the same time, bioinformatics was used to analyze mitochondrial differential proteins by go function enrichment analysis and protein interaction network construction. The expression of some mitochondrial differentially expressed proteins in 5-8F cells of nasopharyngeal carcinoma (NPC) was assayed by siRNA transient interference transwell migration assay. Results: sixteen mitochondrial differential proteins including PRDX3 and SOD2 were identified. Inhibition of PRDX3 expression in 5-8F cells of nasopharyngeal carcinoma (NPC) showed that inhibition of PRDX3 expression in 5-8F cells could enhance cell motility. Go functional enrichment analysis showed that the biological processes were cell response to reactive oxygen species (Ros), hydrogen peroxide metabolism, and regulation of mitochondrial membrane potential. Cell redox dynamic balance and redox activity, the molecular function mainly manifested in redox enzyme activity, cysteine protease inhibitor activity, peroxidase activity, The activity of plasma membrane channel protein and antioxidant were 5 aspects. Through data mining, a network of mitochondrial differential protein interactions was constructed. Based on the hypothesis of tumor-stromal coevolution, the possible molecular mechanisms of 10 mitochondrial differential proteins, such as PRDX3, PRDX6, SOD2, ECH1, SERPINB5, COX5A, PDIA5EIF5AtIIDH3B and PSMC4, in the metastasis of nasopharyngeal carcinoma were preliminarily revealed. It provides experimental evidence for the view that oxidative stress directly promotes tumor metastasis. Conclusion: a total of 16 mitochondrial differentially expressed proteins were identified, and 10 of them were identified as possible molecular mechanisms involved in tumor metastasis. Inhibition of PRDX3 expression in 5-8 F cells of nasopharyngeal carcinoma can improve cell motility. They may be molecular markers of NPC metastasis and play an important role in mitochondrial oxidative stress-induced metastasis of NPC.
【学位授予单位】：中南大学
【学位级别】：博士
【学位授予年份】：2013
【分类号】：R739.63

【相似文献】