Transdifferentiation of human umbilical cord mesenchymal stem cells into cancer-associated mesenchymal stem cells induced by hepatocellular carcinoma HepG-2 supernatant

doi:10.3969/j.issn.2095-4344.2017.01.005

Abstract

Abstract:

BACKGROUND: Tumor microenvironment can recruit and attract different type and differently differentiated mesenchymal stem cells (MSCs) to the tumor growth site, thereby affecting tumor progression.
OBJECTIVE: To investigate the transdifferentiation of human umbilical cord mesenchymal stem cells (hUC-MSCs) into cancer-associated MSCs induced by the supernatant from hepatocellular carcinoma cells HepG-2 culture medium and the effect of induced hUC-MSCs on the proliferation and migration of hepatocellular carcinoma cells HepG-2.
METHODS: (1) Experiment 1: The supernatant from hepatocellular carcinoma cells HepG-2 culture medium was prepared, mixed with equal volume of low glucose DMEM and used to culture hUC-MSCs for 48 hours. The expressions of cancer-associated MSCs proteins and miR-221 in hUC-MSCs were detected. (2) Experiment 2: The supernatant from induced hUC-MSCs culture medium was mixed with equal volume of high glucose DMEM to treat hepatocellular carcinoma cells HepG-2 for 48 hours and then the proliferation and migration of hepatocellular carcinoma cells HepG-2 were observed. (3) Experiment 3: Hepatocellular carcinoma cells HepG-2 were cultured alone or co-cultured with hUC-MSCs for 48 hours and then the capabilities of proliferation and migration of hepatocellular carcinoma cells HepG-2 were observed.
RESULTS AND CONCLUSION: (1) Experiment 1: The protein levels of vimentin and fibroblast activation protein were increased and the expression of miR-221 was upregulated in the hUC-MSCs after treated by the supernatant from hepatocellular carcinoma cells HepG-2 culture medium. (2) Experiment 2: The supernatant from induced hUC-MSCs culture medium promoted hepatocellular carcinoma cells HepG-2 proliferation and migration. (3) Experiment 3: The capabilities of proliferation and migration of hepatocellular carcinoma cells HepG-2 after co-cultured with hUC-MSCs were significantly enhanced. To conclude, these results above declared that the supernatant from hepatocellular carcinoma cells HepG-2 culture medium could induce hUC-MSCs, equipping them with cancer-associated MSC-like phenotype and promoting the proliferation and migration of hepatocellular carcinoma cells HepG-2.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Mesenchymal Stem Cells, Liver Neoplasms, Tumor Microenvironment, Cell Proliferation, Tissue Engineering

CLC Number:

R394.2

Yang Juan, Zheng Sheng, Chen Wen-qin, Liu Jing, Zhang Fan, Wang Yu-bo. Transdifferentiation of human umbilical cord mesenchymal stem cells into cancer-associated mesenchymal stem cells induced by hepatocellular carcinoma HepG-2 supernatant[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(1): 25-31.

Figures/Tables 2

2.1.1 hUC-MSCs经HepG-2培养液上清处理后肿瘤相关蛋白质检测结果对处理前后hUC-MSCs的形态进行镜下观察发现，实验组与对照组细胞形态无明显差差异。Western blot结果提示实验组肿瘤相关间充质干细胞波形蛋白和凋亡调控蛋白水平高于对照组(P < 0.05)，见图1。采用免疫荧光对上述两种肿瘤相关间充质干细胞蛋白质波形蛋白和凋亡调控蛋白水平进行检测，结果提示实验组两种蛋白质的荧光强度明显增强，与Western blot结果一致，见图2。 2.1.2 HepG-2上清处理后hUC-MSCs中的miR-221检测结果实验组miR-221表达明显高于对照组(P < 0.05)，见图3。 2.2 实验2结果 2.2.1 处理后hUC-MSCs对HepG-2增殖能力的影响实验组肝癌HepG-2细胞形成的细胞集落数量较多，单个直径较大，与对照组、空白对照组比较差异有显著性意义(P < 0.05)，对照组与空白对照组比较差异无显著性意义(P > 0.05)，见图4。实验组HepG-2细胞数目明显多于对照组、空白对照组(P < 0.05)，对照组与空白对照组比较差异无显著性意义(P > 0.05)，见图5。 2.2.2 处理后hUC-MSCs对HepG-2迁移能力的影响实验组有更多的HepG-2细胞迁移至Transwell小室膜下层，细胞计数明显高于对照组、空白对照组(P < 0.05)；对照组与空白对照组细胞计数比较差异无显著性意义(P > 0.05)，见图6。此外，细胞形态观察提示3组细胞没有明显差异，见图7。 2.3 实验3结果 2.3.1 共培养后HepG-2增殖能力的变化相较于对照组，实验组的细胞集落数量更多，单个直径更大(P < 0.05)，见图8。从共培养第5天起，实验组细胞增殖速度显著加快，与对照组相比差异有显著性意义(P > 0.05)，见图9。 2.3.2 共培养后HepG-2迁移能力的变化实验组HepG-2迁移细胞明显多于对照组(P < 0.05)，见图10。"

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