舌鳞状细胞癌细胞与正常黏膜细胞微泡的差异蛋白组学研究

目的 通过蛋白组学方法初步分析舌鳞状细胞癌细胞和正常黏膜细胞微泡内蛋白质的差异表达情况，为进一步探讨舌癌复发、转移、扩散机制提供分子生物学依据。方法 体外培养人舌鳞状细胞癌细胞（Tca8113细胞）和人正常黏膜细胞（HOK细胞），获得培养上清液，通过差速离心法分离得到纯化的微泡，采用双向电泳和串联质谱联合分析法寻找差异表达蛋白，通过数据库在线查找差异蛋白的功能。结果 体外培养的Tca8113和HOK细胞均可分泌大量的囊泡状结构物质，这些囊泡状物质经过电子显微镜观察及微泡表面标志物热休克蛋白-70、主要组织相容性复合体Ⅰ类分子的鉴定，证实为微泡；通过差异蛋白组学方法，发现两组微泡中差异表达量在2倍及以上的蛋白差异点有16个，Tca8113细胞微泡上调表达的蛋白质有12个，下调表达的有4个。结论 差异蛋白在微泡形成及癌症进展过程中有非常重要的作用。

Differential proteomics research on exosomes derived from tongue squamous cell carcinoma cells and normal mucosa cells

Objective  This study aimed to explore further the mechanisms of tongue squamous cell carcinoma (TSCC) cell recurrence, metastasis, and diffusion, as well as to establish the experimental basis for the molecular biology research on TSSC. We intend to complete our objective through differential proteomics and preliminary analysis protein expression of exosomes derived from TSCC and normal mucosa cells. Methods  We acquired cultured supernatant fluid in vitro in the laboratory by culturing TSCC (tongue cancer Tca8113 cell line) and human normal mucosa cells (HOK cell line). The exo-somes were separated and purified through differential centrifugation. Furthermore, the different protein expressions were identified through dielectrophoresis and mass spectrometry. The functions of the different protein expressions were identified through an online database search. Results  TSCC and human normal mucosa cells secrete a large amount of capsule bubble structure substances in vitro, as confirmed by electron microscopy and surface markers heat shock protein-70 and major histocompatibility complex class Ⅰ. A total of 16 oral cancer cell-derived exosomes that expressed quantity more than two times, twelve that increased their expression levels, and four that cut their expressions were identified through the differential proteomics research on the two groups. Conclusion  Differential proteins that were verified through the online database serve an important function in exosome formation and in the progress of cancer.