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| AC133和EpCAM在人肺腺癌中的表达及双阳性细胞的分选 |
| 赵振国△,罗福康△,安江宏,段玉忠,孙建国,朱波,陈正堂* |
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| (第三军医大学新桥医院肿瘤科,全军肿瘤研究所,重庆 400037) |
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| 摘要: |
| 目的:通过双重免疫荧光染色法检测人肺腺癌组织标本中AC133以及上皮细胞黏附分子(EpCAM)的表达,并以流式细胞术分选AC133+EpCAM+细胞,为后续人肺腺癌干细胞研究提供实验基础。方法:取人肺腺癌组织标本,冰冻切片,进行双重免疫荧光染色,通过激光共聚焦显微镜检测腺癌组织内AC133和EpCAM的表达。以胶原酶和分散酶将新鲜肺腺癌组织标本制为单细胞悬液,滤除其中的胶原并去除红细胞后,以AC133和EpCAM共同标记单细胞,上流式细胞仪进行分选。结果:在人肺腺癌标本中具有AC133和EpCAM的表达；通过流式细胞术可分选到AC133和EpCAM共表达细胞。结论:验证了人肺腺癌组织中AC133和EpCAM的表达情况；通过流式细胞术分选获得AC133和EpCAM双标记阳性细胞,可为人肺腺癌干细胞的后续研究提供实验基础。 |
| 关键词: AC133 上皮细胞黏附分子 人肺腺癌干细胞；流式细胞术 |
| DOI：10.3724/SP.J.1008.2009.0487 |
| 投稿时间：2009-03-19修订日期：2009-04-27 |
| 基金项目:国家高技术发展研究计划(“863”项目)(2008aa02Z104). |
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| Expression of AC133 and EpCAM in human pulmonary adenocarcinoma and isolation of their double positive cells |
| ZHAO Zhen-guo△,LUO Fu-kang△,AN Jiang-hong,DUAN Yu-zhong,SUN Jian-guo,ZHU Bo,CHEN Zheng-tang* |
| (Department of Oncology,Cancer Center of PLA,Xinqiao Hospital,Third Military Medical University,Chongqing 400037,China) |
| Abstract: |
| Objective:To determine the expression of AC133 and EpCAM in human pulmonary adenocarcinoma by dual immunofluorescent labeling technique and to isolate AC133+EpCAM+ cells by flow cytometry,so as to provide a basis for further investigation of human pulmonary adenocarcinoma stem cells.Methods:The human lung adenocarcinoma tissues were obtained and subjected to cryosection and dual immunofluorescent staining.AC133+EpCAM+cells in human pulmonary adenocarcinoma were identified by using laser confocal microscopy.The fresh adenocarcinoma tissues were prepared into single cell suspension with the collagen and red blood cell removed.AC133 and EpCAM were used to label cells and the AC133+EpCAM+cells were isolated by flow cytometry.Results:AC133+EpCAM+ cells were found in human pulmonary adenocarcinoma and they could be isolated by flow cytometry.Conclusion:The existence of AC133+EpCAM+ cells has been confirmed in the lung adenocarcinoma tissues; the double positive cells can be isolated by flow cytometry,which provides a basis for further investigation of lung cancer stem cells. |
| Key words: AC133 EpCAM human pulmonary adenocarcinoma stem cell flow cytometry |
