CAJ | 학술논문

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.
목적 검측인이선암간세포적화료내약성급미소RNA-200c(miR-200c)적표체.방법 응용류식세포술(FACS)재인이선암세포주PANC-1중분선출CD24+ CD44+ ESA+세포,통과NOD/SCID소서이충이식성류실험증실기종류간세포특성.새서람(MTT)법검측세포대길서타빈적민감성,FACS기술검측세포적항조망능력.실시형광정량취합매련반응(FQ-PCR)법검측세포중miR-200c적표체.결과 인이선암세포주PANC-1중분선출CD24+ CD44+ ESA+세포(0.8％),이충이식성류실험증실기위종류간세포.길서타빈대이선암간세포적반수억제제량(IC50)치위(19.15 ±1.53) μmol/L,명현고우대PANC-1세포적IC50치(0.86 ±0.18) μmol/L (P ＜0.05).재1、10 μmol/L적길서타빈작용후,이선암간세포적조망솔[(0.69±0.09)％、(0.90±0.13)％]명현저우PANC-1세포적조망솔[(60.54±3.73)％、(91.76 ±5.07)％,P＜0.05].이선암간세포중miR-200c적상대표체치(0.15±0.01)현저저우PANC-1세포중miR-200c적상대표체치(1.00±0.09,P＜0.05).결론 이선암간세포구유경강적화료내약성,miR-200c재이선암간세포중표체현저하조,가능삼여료이선암간세포내약적발생.
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.