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目的:探讨micro RNA-200c(miRNA-200c)在胰腺癌干细胞中的表达及作用。方法:应用流式细胞术在人胰腺癌PANC-1细胞中以CD24+CD44+ESA+为标记物分选胰腺癌干细胞,并通过NOD/SCID小鼠移植瘤实验验证其肿瘤干细胞特性;分别用RFQ-PCR法Transwell试验检测PANC-1细胞、胰腺癌干细胞以及转染miRNA-200c前体片段或阴性对照序列的胰腺癌干细胞的miRNA-200c表达与侵袭能力。结果:PANC-1细胞中分选出CD24+CD44+ESA+细胞(占0.8%)具有肿瘤干细胞特性,其在小鼠皮下移植后的移植瘤体积明显大于同期移植的PANC-1细胞[(1 725.14±261.29)mm3 vs.(479.65±99.67)mm3,P<0.05];胰腺癌干细胞中miRNA-200c的表达量明显低于PANC-1细胞(0.15±0.01 vs.1.00±0.09,P<0.05),平均穿膜细胞数明显多于PANC-1细胞[(321±7.62)个vs.(70±16.47)个,P<0.05],但转染miRNA-200c前体片段后,胰腺癌干细胞中miRNA-200c表达量明显升高,平均穿膜细胞数明显减少(P<0.05)。结论:胰腺癌干细胞中miRNA-200c的表达降低,miRNA-200c具有抑制胰腺癌干细胞生长及侵袭力的作用。
Objective: To investigate the expression and function of microRNA-200c (miRNA-200c) in pancreatic cancer stem cells. Methods: Pancreatic cancer stem cells were sorted by CD24 + CD44 + ESA + in human pancreatic cancer PANC-1 cells by flow cytometry. The characteristics of tumor stem cells were verified by the experiment of NOD / SCID mice transplantation. -PCR Transwell assay was used to detect the expression and invasion of miRNA-200c in PANC-1 cells, pancreatic cancer stem cells and pancreatic cancer stem cells transfected with miRNA-200c precursor fragment or negative control sequence. Results: The CD24 + CD44 + ESA + cells (0.8%) were selected as the tumor stem cells in PANC-1 cells. The volume of the transplanted tumor after transplanted subcutaneously in PANC-1 cells was significantly larger than that of PANC-1 cells transplanted in the same period The expression of miRNA-200c in pancreatic cancer stem cells was significantly lower than that in PANC-1 cells (0.15 ± 0.01 vs.1.00 ± 0.09, P <0.05) The average number of transplanted cells was significantly higher than that in PANC-1 cells [(321 ± 7.62) vs. (70 ± 16.47), P <0.05]. However, after transfection of the miRNA-200c precursor fragment, the miRNA- 200c expression was significantly increased, the average number of transmembrane cells was significantly reduced (P <0.05). Conclusion: The expression of miRNA-200c in pancreatic cancer stem cells is decreased, miRNA-200c can inhibit the growth and invasion of pancreatic cancer stem cells.