Xia Hui, Yu Changhai, Zhang Wen, Zhang Baoshi, Zhao Yingnan, Fang Fang
Department of Carciothoracic Surgery, the First Affiliated Hospital of the General Hospital of PLA, Beijing 100048, China.
Zhongguo Fei Ai Za Zhi. 2013 Aug 20;16(8):400-4. doi: 10.3779/j.issn.1009-3419.2013.08.02.
Lung cancer stem cells are the root causes of lung cancer malignant phenotype and potential therapeutic target, the aim of this study is to isolate and characterize the cancer stem cells in the lung adenoearcinomas cell line A549, so as to provide an experimental basis for further stem cell research.
The cancer stem cells were isolated from the lung adenoearcinomas cell line A549 using FACS. And the difference of colony formation, cell proliferation and tumorigenicity in vitro were also tested. The expression of CD133 and ABCG2 were evaluated by RT-PCR and Western blot.
The percentage of SP cells was 5.93% of A549 and 0.32% of A549 after incubation with verapamil. The results showed that there were significantly higher expression of CD133 and ABCG2 on SP cells than that of non-SP cells. And the ability of colony formation, cell proliferation and tumorigenicity in SP cell group were remarkably higher than that in non-SP cell group.
Our results suggested that the cancer stem cells with higher expression of CD133 and ABCG2 can be isolated from the lung adenoearcinomas cell line A549 using FACS and be used in the further research experiments.
肺癌干细胞是肺癌恶性表型的根源及潜在治疗靶点，本研究旨在从肺腺癌A549细胞系中分离并鉴定癌干细胞，为进一步的干细胞研究提供实验依据。
采用荧光激活细胞分选术（FACS）从肺腺癌A549细胞系中分离癌干细胞。并检测其体外集落形成、细胞增殖及致瘤性的差异。通过逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹法（Western blot）评估CD133和ABCG2的表达。
维拉帕米孵育后，A549细胞中侧群（SP）细胞百分比为5.93%，A549细胞中为0.32%。结果显示，SP细胞上CD133和ABCG2的表达明显高于非SP细胞。且SP细胞组的集落形成、细胞增殖及致瘤性能力明显高于非SP细胞组。
我们的结果表明，利用FACS可从肺腺癌A549细胞系中分离出CD133和ABCG2高表达的癌干细胞，并用于进一步的研究实验。
