摘要
目的:通过双重免疫荧光染色法鉴定不同肺组织内是否存在Clara细胞特异性抗原(Clara cell specific antigen,CCA)和肺泡表面活性蛋白C(surfactant protein C,SP-C)共表达的双阳性细胞(double positive cell,DPCs)CCA+SP-C+细胞,为后续开展肺干细胞的相关研究提供实验基础。方法:取成年鼠(小鼠、大鼠)和新生鼠(小鼠、大鼠)的正常肺组织,人的肺鳞癌、腺癌的癌组织和癌旁组织,冰冻切片,进行双重免疫荧光染色,通过激光扫描共聚焦显微镜观察肺组织内是否存在DPCs。每例标本观察时镜下随机选择4~6个视野,计数100~200个肺细胞,得出每百个细胞内DPCs百分率,采用SPSS 11.0统计软件包对数据进行分析。结果:在成年鼠和新生鼠的正常肺组织中均发现了DPCs的存在,在人的肺腺癌组织中也发现了DPCs,而在鳞癌组织中未见DPCs;在人的癌旁组织DPCs数量明显多于相对应的癌组织(P<0.05)。结论:本研究不仅证实了DPCs细胞存在的广泛性,而且在人的肺腺癌组织中发现了DPCs,同时发现癌旁组织内的DPCs明显多于癌组织,为后续进一步分离、纯化DPCs,研究正常肺干细胞和肺癌干细胞的生物学特性提供了实验基础。
Objective： To determine whether there exist the cells expressing both Clara cell antigen （CCA） and surfactant protein C （SP-C） in different lung tissues from mouse, rat, and human by dual immunofluorescent labeling technique. The CCA^＋ and SP-C^＋ cells were named as the double positive cells （DPCs） which may provide the basis for further investigation of lung stem cells. Methods： The normal lung tissues were isolated from adult and newborn mice and rats. The human lung squamous carcinoma and adenocarcinoma tissues and para-cancerous lung tissues were isolated and subjected to cryosection and dual immunofluorescent staining. The DPCs in different lung tissues were identified by using laser confocal microscopy. For each specimen, four to six visual fields were selected and totally 100-200 cells were counted to calculate the percentage of DPCs. The data were analyzed by the SPSS 11.0 software. Results： The DPCs were found in normal lung tissues from adult and newborn mice and rats. DPCs were first discovered in human lung adenocarcinoma tissues but not found in lung squamous carcinoma tissues. Furthermore, the number of DPCs in the paracancerous tissues was significantly increased than the corresponding lung cancer tissues （P 〈 0.05 ）. Conclusion： We not only validate the wide distribution of DPCs in the normal lung tissues, but also discover the existence of DPCs in the lung adenocarcinoma tissues and the number of DPCs in para-cancerous tissues is significantly higher than that in lung cancer tissues. Our study provide the experimental basis for further isolation and identification of DPCs and investigation of the biological features of normal lung stem cells and lung cancer stem cells.
出处
《肿瘤》
CAS
CSCD
北大核心
2008年第1期32-35,共4页
Tumor
基金
国家自然科学基金资助项目(编号:30672076)
关键词
癌
非小细胞肺
荧光抗体技术
抗原
表面
肺表面活性物质相关蛋白质C
肿瘤干细胞
Carcinoma,non-small-cell lung
Fluorescent antibody technique
Antigens, surface
Pulmonary surfactant associated protein C
Tumor stem cells
