摘要
目的 检测人胰腺癌干细胞的化疗耐药性及微小RNA-200c（miR-200c）的表达.方法 应用流式细胞术（FACS）在人胰腺癌细胞株PANC-1中分选出CD24^＋ CD44^＋ ESA^＋细胞,通过NOD/SCID小鼠异种移植成瘤实验证实其肿瘤干细胞特性.噻唑蓝（MTT）法检测细胞对吉西他滨的敏感性,FACS技术检测细胞的抗凋亡能力.实时荧光定量聚合酶链反应（FQ-PCR）法检测细胞中miR-200c的表达.结果 人胰腺癌细胞株PANC-1中分选出CD24^＋ CD44^＋ ESA^＋细胞（0.8％）,异种移植成瘤实验证实其为肿瘤干细胞.吉西他滨对胰腺癌干细胞的半数抑制剂量（IC50）值为（19.15 ±1.53） μmol/L,明显高于对PANC-1细胞的IC50值（0.86 ±0.18） μmol/L （P ＜0.05）.在1、10 μmol/L的吉西他滨作用后,胰腺癌干细胞的凋亡率[（0.69±0.09）％、（0.90±0.13）％]明显低于PANC-1细胞的凋亡率[（60.54±3.73）％、（91.76 ±5.07）％,P＜0.05].胰腺癌干细胞中miR-200c的相对表达值（0.15±0.01）显著低于PANC-1细胞中miR-200c的相对表达值（1.00±0.09,P＜0.05）.结论 胰腺癌干细胞具有更强的化疗耐药性,miR-200c在胰腺癌干细胞中表达显著下调,可能参与了胰腺癌干细胞耐药的发生.
Objective To investigate the sensitivity of human pancreatic cancer stem cells to chemotherapy and the expression of microRNA （miRNA,miR）-200c in cancer stem cells.Methods CD24^＋ CD44^＋ ESA^＋ cells were sorted from PANC-1 cell line by fluorescence-activated cell sorter （FACS）.The stem like properties of this subpopulation were assessed by non-obese diabetic （NOD）/ severe combined immunodeficiency （SCID） xenograft transplantation experiment.Sensitivity to gemcitabine and apoptosis ratio of pancreatic cancer stem cells and PANC-1 cells were detected by methyl thiazol tetrazolium （MTT） assay and FACS respectively.The real-time fluorescent quantitative polymerase chain reaction （FQ-PCR） was used to detect miR-200c expression in pancreatic cancer stem cells and PANC-1 cells.Results CD24^＋ CD44^＋ ESA^＋ cells （0.8%） were isolated in PANC-1 cells.NOD/SCID xenografl transplantation experiment confirmed that the sub-group had the characteristics of cancer stem cells.The half inhibition concentration （IC50） of gemcitabine was significantly higher in pancreatic cancer stem cells group [（19.15 ± 1.53） μmol/L] than in PANC-1 cells group [（0.86 ± 0.18） μmol/L] （P 〈 0.05）.Mter the interference of gemcitabine （1,10 μmol/L）,the apoptosis ratio was significantly lower in pancreatic cancer stem cells group [（0.69 ±0.09）% and （0.90 ±0.13）%] than that in PANC-1 cells group [（60.54 ± 3.73） % and （91.76 ± 5.07） %] （P 〈 0.05）.The relative miR-200c expression level in cancer stem cell line [（0.15 ±0.01）] was significantly lower than that in PANC-1 cell line [（1.00 ± 0.09）] （P 〈 0.05）.Conclusion The expression of miR-200c was significantly reduced in pancreatic cancer stem cells which were significantly chemo-resistant,and miR-200c may play an important role in the chemo-resistance of pancreatic cancer stem cells.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2015年第5期1054-1056,共3页
Chinese Journal of Experimental Surgery
