【摘要】目的 探讨应用激光捕获显微切割(laser capture microdissection, LCM)技术获取高同质性肺癌组织细胞及正常对照肺泡上皮细胞的方法及可行性。方法 将新鲜肺癌组织标本及其配对正常组织常规制备8μm冰冻切片，采用改良的HE方法染色;应用LCM技术大宗切割肺鳞癌、腺癌及正常对照肺泡上皮细胞;选择性获取同质肺癌细胞和配对正常细胞。结果 通过LCM技术可以准确、快速地分离癌细胞、间质及其他坏死组织，得到同质性不低于95%的肿瘤细胞和正常细胞。按照Duration 15.5s的激光脉冲频率和平均8000 shots条件，既可保证每个帽子的细胞收集数量，也能够有效地缩短实验时间，提高实验可控性，从而使操作过程标准化。结论 应用LCM技术可以成功获取同质肺癌细胞和正常细胞。其所需标本量少、组织同质性高、操作过程稳定、可控性好，是肺癌分子生物学及蛋白质组学研究有价值的样品制备方法。
【关键词】 激光捕获显微切割;肺癌组织;蛋白质组学
Application of laser capture microdissection to capture pure cells in lung cancer and their paired normal lung tissues
Tian Yingxuan, Nan Yandong, Yang Shuanying
(Department of Respiratory Medicine, the Second Affiliated Hospital, Medical School of Xian Jiaotong University, Xian 710004, China)
ABSTRACT: Objective Microheterogeneity is an important problem in molecularly biological and proteomic research on malignancies. The goal of this study was to investigate the method to purify targeted cells in lung cancer and its paired normal tissues for proteomic study on lung cancer applying laser capture microdissection (LCM). Methods LCM technique was emploged to obtain the cells of lung cancer tissues and their paired normal tissues from frozen sections. Results LCM can be applied to quickly and precisely obtain pure targeted cells subgroup or single cell without interstitium under the microscope. Conclusion LCM can tackle the problem of tissue heterogeneity in molecular analysis successfully. These results indicate that LCM has a potential as a tool in lung cancer proteomic research.
KEY WORDS: laser capture microdissection; lung cancer tissue; proteomics
蛋白质组学是当今肿瘤学研究领域中的一个热点，而“差异”蛋白质组学又是蛋白质组学研究的一个突破口。在有关“差异”的研究中，若原始样品不是分别来自同一类型，或样品存在着杂质蛋白的污染，那么所得实验数据的准确性和说服力必然会受到影响[1]。众所周知，人体组织器官是多种细胞的复合，肿瘤组织的不均一性是肿瘤细胞的重要特征。因此，如何从复杂、不均一的样品中获得理想的高同质性的组分成为实验中亟需解决的主要问题之一。激光捕获显微切割(laser capture microdissection, LCM)技术的出现为上述问题提供很好的解决方案。该方法的突出特点是能在目视下从样品中特异挑选同类细胞乃至单一细胞;从而剔除间质细胞和坏死
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