激光捕获显微切割技术纯化的肺鳞癌原发灶与淋巴结转移灶肿瘤细胞的比较蛋白质组学

Comparative proteome analysis of laser capture microdissection for purified primary tumor and lymph node metastatic tumor in human lung squamous carcinoma

目的:筛选肺鳞癌淋巴结转移相关的蛋白质。方法:采用激光捕获显微切割技术(lasercapture microdissection,LCM)分别从肺鳞癌原发灶组织和匹配的淋巴结转移灶癌组织中切割纯化鳞癌细胞;应用二维凝胶电泳(two-dimensional electrophoresis,2-DE)分离LCM纯化细胞的蛋白质;图像分析识别差异表达的蛋白质点;基质辅助激光解吸电离飞行时间质谱鉴定差异蛋白质点,Western印迹验证差异蛋白Rho-GDIα在LCM纯化的肺鳞癌原发灶肿瘤细胞(lung primary tumor cells,LPTC)和匹配的淋巴结转移灶肿瘤细胞(lymph node metastatic tumor cells,LNMTC)中的表达。结果:建立了LCM纯化的LPTC和匹配的LNMTC的2-DE图谱,质谱鉴定了22个差异蛋白质,与LPTC相比,8个蛋白质在LNMTC中表达明显增高。结论:22个差异蛋白可能与肺鳞癌淋巴结转移有关,为筛选肺鳞癌转移标志物奠定了基础。

Objective To search for lymph node metastasis-associated proteins in human lung squamous carcinoma（hLSC）.Methods Laser capture microdissection（LCM）was used to purify the target cells from lung primary tumor and matched lymph node metastatic tumor in hLSC.Two-dimensional gel electrophoresis（2-DE）was performed to separate the total proteins of microdissected tumor cells from lung primary tumor and matched lymph node metastatic tumor.PDQuest software was applied to analyze 2-DE images.Differential protein spots between the two types of tissues were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry（MALDI-TOF-MS）.The expression of Rho-GDIα,one of the differential proteins,in the microdissected lung primary tumor cells（LPTC）and matched lymph node metastatic tumor cells（LNMTC）was detected by Western blot.Results In the present study,2-DE patterns of microdissected LPTC and LNMTC were established,and 22 differential proteins in the above two tissues were identified,of which 14 were down-regulated in LNMTC and 8 were up-regulated in LNMTC.Conclusion The 22 differential proteins may play some roles in the process of lymph node metastasis in hLSC,and the data provide new clues for metastasis-associated biomarker screen and mechanism of hLSC.