胰腺癌肿瘤干细胞样侧群细胞的分离鉴定

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HoechsG3342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.结论 SP细胞可能是胰腺癌干细胞的候选细胞之一.

Isolation and identiffcation of cancer stem-like side population cells in pancreatic cancer

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 cels and non-SP ceils by clone formation assay and NOD/SCID xenograft transplantation experiment.The differentiation ability of SP ceils was studied by flow cytometry reanalysis of SP-derived tumors and cultured SP ceils.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 ceUs were demonstratcd 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 ceHs 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 100-fold compared with non-SP cells.and had the capacity to generate nontumorigenic non-SP cells through asymmetrical cell division in vivo.Conclusion SP phenotype iS a potential marker for pancreatic cancer stem cells.