LCM纯化的鼻咽癌间质和正常鼻咽间质的定量蛋白质组学研究

Quantitative Proteomics Analysis of LCM Purified Stroma of Nasopharyngeal Carcinoma and Normal Nasopharyngeal Mucosa

间质在肿瘤发生发展中的作用越来越受到重视.为寻找与鼻咽癌(nasopharyngeal carcinoma,NPC)发生发展相关的特异性间质蛋白,采用激光捕获显微切割技术(laser capture microdissection,LCM)纯化鼻咽癌间质和正常鼻咽黏膜间质,荧光差异双向凝胶电泳(fluorescent two-dimensional difference gel electrophoresis 2-D,DIGE)结合质谱技术分离鉴定间质相关蛋白.Westernblot及免疫组织化学技术验证了其中3个差异蛋白(CapG、L-plastin和S100A9),证实了2D-DIGE结果的可靠性.建立了LCM纯化的鼻咽癌间质和正常鼻咽间质的荧光差异蛋白表达图谱,高通量筛选与肿瘤发生相关的间质蛋白,共得到34个有统计学意义的蛋白质点,质谱鉴定得到20个差异蛋白.研究结果提示:这些差异表达的蛋白质将有助于阐明鼻咽癌细胞和周围间质的关系.对间质蛋白功能的进一步研究,将有助于解析间质在肿瘤发生中的作用机制,并为从间质途径寻找肿瘤治疗靶标提供新思路.

The mechanism of how stroma plays an important role in tumor carcinogenesis is now a hotspot. To delineate the features of stromal protein between nasopharyngeal carcinoma （NPC） and normal nasopharyngeal mucosa （NNM）, laser capture microdissection （LCM） was performed to purify stromal cells fi＇om the NPC and NNM, respectively. The protein expressed profiles of the stroma of NPC and NNM were compared using fluorescent two-dimensional difference gel electrophoresis （2D-DIGE） and 34 differential protein spots between tumor stroma and normal stroma were chosen to be identified by mass spectrometry （MS）. A total of 20 differential proteins were identified, and three differential proteins （CapG, L-plastin and S100A9） were selectively further validated by Western blotting and immunohistochemical analysis to confirm the results of 2D-DIGE. 2D-DIGE patterns of the stroma of NPC and NNM were established for the first time, the results suggested that differentially expressed proteins in the stroma of NPC and NNM may be useful for understanding the relationship between NPC cells and their surrounding microenvironment. Further studying of these proteins will be helpful to elucidate the mechanisms of NPC carcinogenesis and provide new thoughts on therapy of NPC through stroma.