蛋白质组学技术下人皮肤鳞状细胞癌细胞和基底细胞癌细胞中差异蛋白质分析

摘要

目的 研究分析蛋白质组学技术下人皮肤鳞状细胞癌细胞和基底细胞癌细胞中的差异蛋白质.方法 选取购自美国菌种保藏中心的皮肤鳞状的细胞癌的细胞系A-431和皮肤基底的细胞癌的细胞系TE354.T, 首先对细胞进行培养, 等到细胞生长到培养瓶面的面积91％时候, 提取样品的蛋白, 使用Bradford方法检测样品蛋白蛋白质的浓度;通过IPG预制的干胶条重水化, 进行等电聚焦电泳, 并制作11％聚丙烯酰胺凝胶, 而后平衡IPG的胶条, 随后进行第二向的垂直板的SDS-PAGE电泳, 使用考马斯亮兰的染色法对交替进行染色, 使用扫描仪对图像进行扫描, 得到样品蛋白的双向凝胶点用的图谱, 分析图谱找到蛋白质点的差异.结果 鳞状细胞系 (A-431) 的蛋白液在595 nm波长内吸光度为1.104, 带入方程计算得出蛋白质的浓度是1150.7μg/ml;基底细胞系 (TE.354.T) 在595 nm波长内吸光度为0.898, 带入方程计算得出蛋白质的浓度是936.1μg/ml.对2种细胞系蛋白双向凝胶电泳的图谱对比分析, 在2组凝胶内共有19个蛋白质的斑点表达, 差异显著, 具有统计学意义 (P＜0.05), 其中在鳞状癌细胞内表达下降有2个蛋白, 在鳞状癌细胞内表达上升有13个蛋白, 基底的癌细胞内没有检测出有4个蛋白.结论 基底细胞系与鳞状细胞系内提取蛋白样品进行的双向电泳图谱内有差异存在, 这些差异的蛋白质是影响2种肿瘤在转移方面存在差异的主要因素.%Objective To analyze differential proteins in human skin squamous cell carcinoma cells and basal cell carcinoma cells under proteomics techniques.Methods The cell line A-431 of the cutaneous squamous cell carcinoma and the cell line TE354.T of the basal cell carcinoma were purchased from the American Type Culture Collection, and the cells were first cultured until the cells had grown to the surface of the flask.At the time of 91％, the protein of the sample is extracted, the protein protein concentration of the sample is detected using the Bradford method, the IPG prefabricated dry strips are rehydrated, isoelectric focusing is performed, and an 11％polyacrylamide gel is prepared, and then the IPG strip is equilibrated Then, the second-direction vertical plate was subjected to SDS-PAGE electrophoresis and stained alternately using the staining method of Coomassie brilliant blue.The image was scanned using a scanner to obtain a map of the two-dimensional gel points of the sample protein.The analysis spectrum was found.Differences in protein spots.ResultsThe absorbance of the protein solution of the squamous cell line (A-431) was 1.104 at a wavelength of 595 nm, the protein concentration was 1150.7μg/ml calculated by the equation, and the basal cell line (TE.354.T) was at a wavelength of 595 nm.The absorbance was 0.898, and the equation was calculated to find that the protein concentration was 936.1μg/ml.Two-dimensional gel electrophoresis analysis of two kinds of cell lines showed that there were 19 protein spots in two groups of gels, and the difference was statistically significant (P＜0.05), and the expression in squamous carcinoma cells was decreased.There were two proteins, 13 proteins in squamous cells and four proteins in the basal cells.Conclusion There are differences in the two-dimensional electrophoresis patterns of proteins extracted from basal cell lines and squamous cell lines.These differential proteins are the main factors affecting the differences in the metastasis of the two tumors.