Background and objective: 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.
Methods: 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.
Results: 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.
Conclusions: 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.
背景与目的 肺癌干细胞是肺癌恶性表型的根源和潜在的治疗靶点，从人肺癌A549细胞株中分离肺癌干细胞，观察特异性干细胞标志物分子的表达，为进一步的干细胞研究提供试验基础。方法 接种肺癌A549细胞株，经流式细胞术，特异性筛选分离肺癌干细胞，观察克隆形成能力、细胞增殖能力和体外致瘤能力的差别，并分别用RT-PCR和Western blot的方法分析干细胞标志物分子CD133和ABCG2的表达。结果 经过流式细胞仪成功分选了人肺腺癌A549细胞系SP细胞亚群，结果表明此SP细胞亚群约占A549细胞总数的5.93%，经维拉帕米处理后Hoechest33342阴性/弱阳性细胞含量下降为0.32%，SP细胞克隆形成能力，细胞增殖能力和体外致瘤能力均明显高于非SP细胞。RT-PCR和Western blot结果发现，筛选分离的肺癌SP细胞群高表达干细胞标志物分子CD133和ABCG2。结论 通过流式细胞术可以筛选分离高表达CD133和ABCG2分子的肺癌干细胞，可用于进一步的研究中。