皮肤鳞状细胞癌与基底细胞癌差异蛋白质组学研究

发布时间：2018-01-06 13:09

本文关键词：皮肤鳞状细胞癌与基底细胞癌差异蛋白质组学研究　出处：《郑州大学》2014年硕士论文　论文类型：学位论文

【摘要】：背景与目的鳞状细胞癌和基底细胞癌均为常见的皮肤恶性肿瘤，它们都起源于表皮或皮肤附属器，具有相近的好发部位和发病机制，并且两种疾病在发病部位的局部也有相似的临床表现。但在肿瘤发生转移的情况上，两种疾病有着较大差异。蛋白质组学（proteomics）是指将细胞或组织中的蛋白质组作为整体进行研究，分析蛋白质组中各蛋白的成分组成以及相互之间的的影响。蛋白质组学主要是对蛋白质组的蛋白质构成、差异蛋白质组学研究以及在生物活动中各蛋白质之间发挥的作用这三个方面进行主要的研究。这次实验应用蛋白质组学的相关技术对人皮肤鳞状细胞癌细胞和基底细胞癌细胞中所提取的蛋白质进行分析，找出其中存在差异的蛋白质，并对部分差异蛋白进行功能验证。这将对皮肤鳞状细胞癌、基底细胞癌的早期疾病诊断、早期发生及转移等肿瘤标志物的鉴定、肿瘤治疗特异性位点的筛选，以及新的临床治疗方案的制定均会有重要的意义。方法 1细胞的培养与样品蛋白的提取本实验选择人皮肤鳞状细胞癌A-431细胞系和人基底细胞癌TE-354.T做为肿瘤细胞模型进行常规细胞培养。分别取对数生长期的两种细胞，待细胞贴壁生长至培养瓶培养面面积的90%时，进行样本蛋白提取。并应用Bradford法对样品蛋白浓度进行测定。 2样品蛋白的双向凝胶电泳通过IPG干胶条重水化、等电聚焦电泳、平衡IPG胶条、垂直向SDS-PAGE电泳、颜色等步骤分别对两种样品蛋白质进行蛋白分离，每个样品分别制3块双向电泳胶板，并应用图像扫描仪进行扫描，获得两种蛋白样品的双向凝胶电泳图谱。用图像分析软件对双向凝胶电泳图谱进行分析，找出差异蛋白质点。结果 1两种细胞系提取蛋白的双向电泳蛋白图谱大眼管基本一致，各个蛋白质着色点基本都聚集在等电点（Ip）为pH4~7的范围内；分子量（Ms）大多在15kDa~150kDa的范围内。 2通过对在皮肤鳞状细胞癌细胞系和皮肤基底细胞癌细胞系中提取样品蛋白的双向凝胶电泳胶板的分析，其中有16.8%的蛋白质点存在差异，其中13个蛋白在鳞状细胞癌细胞中表达上调；和2个在鳞状癌细胞细胞表达下调的蛋白质。4个蛋白点在基底细胞癌组中未检测出。结论 1皮肤鳞状细胞癌细胞系和皮肤基底细胞癌细胞系中提取样品蛋白的双向凝胶电泳图谱中存在部分差异。皮肤鳞状细胞癌所含蛋白质组分较基底细胞癌的复杂。 2分子量和等电点分别为（22.32，3.95）、（27.48，4.3）、（58.96，7.45）、（64.8，5）、（121.8，，5.84）、（135.13，6.44）等蛋白质的表达差异可能是造成鳞状细胞癌和基底细胞癌在肿瘤转移方面有较大差异的主要影响因素。
[Abstract]:Background and Purpose Squamous cell carcinoma and basal cell carcinoma are common malignant tumor of the skin , all of them originate from the epidermis or the skin accessory , have similar good hair parts and pathogenesis , and the two diseases have similar clinical manifestations at the site of the disease . But in the case of tumor metastasis , the two diseases have a large difference . Proteomics refers to the study of the composition of each protein in the protein group and its influence on each other . Proteomics is mainly used to study the structure of protein in the proteome , the differential proteomic study and the role of the proteins in the biological activity . In this experiment , the proteins extracted from human skin squamous cell carcinoma cells and basal cell carcinoma cells were analyzed by using the technique of Proteomics . It was found that there were different proteins and functionally validated some of the difference proteins . This would be of great significance to the identification of tumor markers such as squamous cell carcinoma of skin , early diagnosis of basal cell carcinoma , early occurrence and metastasis , screening of tumor treatment specific sites , and the development of new clinical treatment programmes . method Human skin squamous cell carcinoma A - 431 cell line and human basal cell carcinoma TE - 354.T were used as tumor cell model to carry out normal cell culture . The two - way gel electrophoresis of two kinds of sample proteins was carried out by two - way gel electrophoresis of two sample proteins , such as rehydration , isoelectric focusing , SDS - PAGE electrophoresis , color and so on . The two - way gel electrophoresis map of two kinds of protein samples was obtained by scanning with image scanner . The two - way gel electrophoresis map was analyzed by image analysis software to find out the difference protein spots . Results The isoelectric point ( Ip ) of each protein staining point is within the range of pH 4 - 7 , and the molecular weight ( Ms ) is mostly in the range of 15 kDa to 150 kDa . 2 . Analysis of two - way gel electrophoresis gel electrophoresis for the extraction of sample proteins in cancer cell lines and basal cell carcinoma cells of the skin . There were 16.8 % of the protein spots , 13 of which were up - regulated in squamous cell carcinoma cells , and 2 proteins down - regulated in squamous cell carcinoma cells . Four protein spots were not detected in basal cell carcinoma . Conclusion There are some differences in the two - way gel electrophoresis of the sample protein extracted from the cancer cell line of skin squamous cell carcinoma and basal cell line of skin . The protein component of squamous cell carcinoma of skin is more complex than basal cell carcinoma . The molecular weight and isoelectric point were ( 22.32 , 3.95 ) , ( 27.48 , 4.3 ) , ( 58.96 , 7.45 ) , ( 64.8 , 5 ) , ( 121.8 , 5.84 ) , ( 135.13 , 6.44 ) and so on .

【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R739.5

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