CAJ | 학술논문

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。
목적：통과단백조학방법초보분석설린상세포암세포화정상점막세포미포내단백질적차이표체정황，위진일보탐토설암복발、전이、확산궤제제공분자생물학의거。방법체외배양인설린상세포암세포（Tca8113세포）화인정상점막세포（HOK세포），획득배양상청액，통과차속리심법분리득도순화적미포，채용쌍향전영화천련질보연합분석법심조차이표체단백，통과수거고재선사조차이단백적공능。결과체외배양적Tca8113화HOK세포균가분비대량적낭포상결구물질，저사낭포상물질경과전자현미경관찰급미포표면표지물열휴극단백-70、주요조직상용성복합체Ⅰ류분자적감정，증실위미포；통과차이단백조학방법，발현량조미포중차이표체량재2배급이상적단백차이점유16개，Tca8113세포미포상조표체적단백질유12개，하조표체적유4개。결론차이단백재미포형성급암증진전과정중유비상중요적작용。
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.