人肺腺癌A549细胞系SP细胞亚群的分选及初步探讨

ISOLATION AND ANALYSIS OF SIDE POPULATION CELLS IN HUMAN LUNG ADENOCARCINOMAS CELL LINE A549

目的:探讨一种简单有效的分离肺癌干细胞的方法。方法:用免疫组化方法检测人肺腺癌A549细胞系中ABCG2蛋白的表达情况;用流式细胞学方法分选A549细胞系中sidepo pulationcells(SP细胞)及非SP细胞亚群,并对该两亚群细胞的分化功能进行检测;用RT-PCR方法检测两亚群细胞ABCG2的表达。结果:免疫组化方法检测A549细胞系中ABCG2蛋白的阳性表达率为2%。流式方法成功分选得到SP细胞及非SP细胞亚群,其中SP细胞含量约为A549活细胞总量的1.09%,经拮抗剂Verapamil处理后下降为0.20%,两亚群细胞经培养后重新上机检测,发现SP细胞亚群可产生SP细胞及非SP两种细胞,而非SP细胞只能产生非SP细胞。SP细胞表达ABCG2,非SP细胞则不表达。结论:人肺腺癌A549细胞系SP亚群细胞富集了肺癌干细胞。通过流式细胞仪分选肺腺癌SP细胞亚群是分离肺腺癌干细胞的有效方法。

Objective：To set up one simple but efficient method of isolation lung cancer stem cells. Methods：The protein expression of ABCG2 in human lung adenocarcinomas cell line A549 was detected by immunohistochemistry. SP and NSP cells in cell line A549 were isolated by FACS respectively, and their differentiation was analysed. ABCG2 expression in the tow cell subsets was detected by RT PCR. Results：The positive rate of ABCG2 in cell line A549 was 2% by immunohistochemistry. SP and NSP cells were isolated by FACS, the percentage of SP cells was 1.09% in all of the alive cell A549, and decreased to 0.20% after treated by Verapamil. SP cells could produce both SP and NSP cells, but NSP cells could only produce NSP cells. Conclusion..SP cells in human lung adenoeareinomas cell line A549 enrich tumor stem cells. An effective way to get lung adenocarcinomas stem cells is to isolate SP cells by FACS.