摘要
目的分离鉴定胰腺癌中的侧群（SP）细胞亚群。方法应用Hoechst33342染色，流式细胞仪检测5个胰腺癌细胞系及3个原代培养的临床胰腺癌标本中sP细胞的含量。以PANC-1为例，通过平板克隆形成试验和NOD-SCID小鼠异种移植成瘤实验比较sP细胞与non-SP细胞的克隆形成能力及成瘤能力，通过对体外培养的SP细胞和SP细胞衍生肿瘤的Hoechst33342复染SP再分析判断其是否具有分化潜能。结果除了BXPC-3，其他胰腺癌细胞系及原代培养标本都存在Verapamil敏感的SP细胞。SP细胞与non—SP细胞比较具有较高的克隆形成能力[（43．67±3．10）％比（8．33±1．63）％，P〈0．01]，并且能够分化产生non—SP细胞并维持自身SP细胞的比例在一个较稳定的水平。SP细胞的成瘤能力是non-SP细胞的100倍以上。而且SP细胞在体内亦可发生不对称分裂生成SP细胞和non—SP细胞。
Objective To isolate and identify side population （SP） cells in pancreatic cancer. Methods We detected the number of SP cells in 5 pancreatic cancer cell lines and 3 clinical samples by Hoechst33342 dyeing and FACS analysis. The clone formation efficiency and tumorigenicity were compared between SP cells and non-SP cells by clone formation assay and NOD/SCID xenograft transplantation experiment. The differentiation ability of SP cells was studied by flow cytometry reanalysis of SP-derived tumors and cultured SP cells. Results All cell lines and clinical samples were found to exhibit verapamilsensitive SP cells except BXPC-3. Using SP cells sorted from PANC-1 as a model, SP cells were demonstrated to exhibit higher colony-formation ability than non-SP cells [ （ 43.67 ± 3. 10 ） % vs （ 8.33 ± 1.63 ） % ,P 〈 0.01 ]. And flow cytometry reanalysis indicated SP cells could generate both SP and non-SP cells with a fraction size comparable with the original population. In addition, SP cells were enriched in tumorigenicity by at least lO0-fold compared with non-SP cells, and had the capacity to generate nontumorigenie non-SP cells through asymmetrical cell division in vivo. Conclusion SP phenotype is a potential marker for pancreatic cancer stem cells.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2009年第1期58-60,共3页
Chinese Journal of Experimental Surgery
基金
基金项目：国家自然科学基金资助项目（30571817）
关键词
胰腺癌
肿瘤干细胞
鉴定
Pancreatic carcinoma
Tumor stem cells
Identification
