Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (49): 9179-9185.doi: 10.3969/j.issn.2095-4344.2012.49.011
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Liang Shuo1, 2, Jiao Hong-liang 3, Chi Lian-kai1, Shi Xin-yi1, Liang A-ming3, Tian Yi3, Han Jiao-ling1, Ma Shan-shan1, Yang Bo3, Guan Fang-xia1, 4
Received:
2012-03-13
Revised:
2012-05-21
Online:
2012-12-02
Published:
2013-01-16
Contact:
Guan Fang-xia, M.D., Professor, Doctoral supervisor,
Department of Bioengineering, Zhengzhou University, Zhengzhou 450001
About author:
Liang Shuo★, Master, Department of Bioengineering, Zhengzhou University, Zhengzhou 450001, Henan Province, China; Biotherapy Center, the First Affiliated Hospital of Henan University of Science and Technology, Luoyang 471003, Henan Province, China shuoguo005@126.com
CLC Number:
Liang Shuo, Jiao Hong-liang,Chi Lian-kai, Shi Xin-yi, Liang A-ming, Tian Yi, Han Jiao-ling, . Human umbilical cord mesenchymal stem cells inhibit glioma growth[J]. Chinese Journal of Tissue Engineering Research, 2012, 16(49): 9179-9185.
2.5 hUC-MSCs对C6细胞Wnt/β-连环蛋白信号通路蛋白表达的影响 Wnt/β-连环蛋白信号转导途径在肿瘤的发生发展中起着至关重要的作用[19-20]。Western blotting结果显示,在hUC-MSCs条件培养液作用下,C6细胞中β-连环蛋白和c-Myc蛋白表达水平下调,见图 4。尽管25%CM组β-连环蛋白和c-Myc表达水平有减小的趋势,但与对照组相比没有明显的变化。然而,随着条件培养液浓度的增加,50%和100% CM组中β-连环蛋白和c-Myc蛋白表达明显下降。因此,结果说明,hUC-MSCs条件培养液中的一些可溶性因子可能参与了抑制C6细胞中Wnt信号途径的过程。"
结果显示,Dickkopf-1蛋白的分泌水平与hUC-MSCs条件培养液的浓度呈正相关。随着条件培养液浓度的增加,Dickkopf-1的分泌量逐渐增多。 2.7 抗体中和效应 见图6。为了进一步证实Dickkopf-1是间充质干细胞抑制细胞增殖的一个关键因子,实验使用了抗Dickkopf-1的抗体来中和hUCMSCs-CM,然后用于处理C6细胞。结果发现,用兔抗人Dickkopf-1多克隆抗体预处理的hUCMSCs-CM失去了下调C6细胞中β-连环蛋白和 c-Myc表达水平的能力,见图6A。同时,CCK-8结果显示,Dickkopf-1中和部分地减弱了hUCMSCs-CM抑制C6细胞增殖的能力,见图6B。流式细胞仪结果显示用hUCMSCs-CM处理的C6细胞在G0/G1期的百分比由54.37%增加到66.48%,见图6C。然而,当Dickkopf-1的活性被抗Dickkopf-1的抗体部分中和后,C6细胞在G0/G1期的百分比由66.48%减少到60.06%,见图6C。这些数据说明,Dickkopf-1的中和作用部分地减弱了hUCMSCs-CM对C6细胞生长的抑制作用,提示了hUCMSCs-CM中的Dickkopf-1具有抑制C6细胞生长的作用。"
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