涎腺腺样囊性癌高低转移细胞系mRNA及蛋白质表达谱差异研究  

Comparison of mRNA and Protein Expression Profiles Between Adenoid Cystic Carcinoma High and Low Metastatic Cell Lines *

作　　者：杨捷琳[1] 朱乃硕[1] 王颖[1] 关晓峰[2] 郑兆鑫[1] 

机构地区：[1]复旦大学生命科学学院病毒及分子免疫实验室,上海200433 [2]中国医科大学口腔医学院,沈阳110001

出　　处：《生物化学与生物物理进展》2004年第4期313-321,共9页Progress In Biochemistry and Biophysics

基　　金：国家自然科学基金资助项目 ( 3 0 0 70 816)~~

摘　　要：转移性和侵袭性是恶性肿瘤的重要特征 ,与基因和蛋白质水平上一系列改变密切相关 .应用mRNA抑制性消减杂交技术和双向电泳结合肽质量指纹分析技术 ,对涎腺腺样囊性癌高、低转移细胞系的mRNA和蛋白质表达谱的差异性进行比较研究 .实验结果显示 ,在抑制性消减杂交中 ,分别以两个细胞系为测试子 ,共获得差异片段 34个 ,其中高转移株细胞中高表达的基因序列有 6个 ,低转移细胞系中有 2 8个 ,其中包括两个新的表达序列标签 (EST) .对这些基因序列 ,进一步以RNA斑点杂交对这些基因的表达状况进行验证并排除假阳性结果 ,结果发现 ,32个基因在mRNA水平上有不同程度的表达量改变 ,改变趋势与消减杂交结果一致 .以蛋白质等电聚焦结合SDS 聚丙烯酰胺凝胶电泳的双向电泳技术获得的总蛋白质分离 ,经PDQuest2DE软件分析结果表明 ,高转移细胞系表达谱的平均蛋白质点数为 (978± 38) ,低转移细胞系的平均蛋白质点数为 (996± 2 7) .其中高转移细胞与低转移细胞相比 ,其蛋白质点有 35 5个未被匹配 ,低转移细胞相比高转移细胞有 2 2 2个未被匹配 .对其中 10个差异较明显的蛋白质点 ,进一步进行基质辅助激光解吸电离飞行时间质谱 (MALDI TOF MS)测定肽质量指纹图谱 ,用Peptident软件对SWISSPROT数据库比较分析 ,结果显示 。The molecular genesis of a tumor is a multi-step and complex process in which many factors are involved such as oncogenes, tumor suppressor genes and MHC-I etc. Metastasis is the essential character causing mortality from such tumors and little is currently known about mechanisms underlying cancer metastasis. As a step toward understanding the complex differences between high metastatic and low metastatic cells, metastasis-associated gene expression patterns and proteomes were separated and identified by suppression subtraction hybridization and comparative proteomics analysis between high metastatic cell AccM and low metastatic cell Acc2. Although extensive similarity was noted between the expression profiles of the two cell lines, twenty-eight genes highly expressed were obtained in tester, low metastatic cells, compared with their low expression in driver, high metastatic cells while six genes in reverse hybridization. These function of genes are related to cell proliferation, differentiation or cell metabolism while several novel EST sequences which function are unknown. Two novel genes were ACC metastasis-associated RNH ACC metastasis-associated suspected protein. Meanwhile, ten proteins were randomly selected which are expressed different qualitatively or quantitatively and identified by mass fingerprinting (MALDI-TOF-MS). These proteins are stratifin, symplekin, B7, TPMsk1 tropomyosin isoforms, serologically defined breast cancer antigen NY-BR-20 etc. Intriguingly, most identified candidate proteins have been shown to be somehow associated with distinct aspects of tumor metastasis such as cell communication protein, motility, adhesion, invasion and tumor immunity associated protein, etc. These results suggest that attaining the ability to metastasize is related to the changed expression of these genes or protein. These data provide insight into the extent of expression differences underlying metastasis-related genes that may prove useful as diagnostic or prognostic markers.

关 键 词：涎腺腺样囊性癌 抑制性消减杂交 双向电泳 肿瘤转移 基质辅助激光解吸电离飞行时间质谱 

分 类 号：Q279[生物学—细胞生物学] R73-37[医药卫生—肿瘤]