Histone deacetylase inhibitors decrease proliferation potential and multilineage differentiation capability of human mesenchymal stem cells
Histone deacetylase inhibitors decrease proliferation potential and multilineage differentiation capability of human mesenchymal stem cells
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DOI：
10.1111/j.1365-2184.2009.00633.x
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发表时间：
2009-12-01
影响因子：
8.5
通讯作者：
Kang, K-S.
中科院分区：
文献类型：
--
作者：
Lee, S.;Park, J-R.;Kang, K-S.
目的：组蛋白脱乙酰酶(HDAC)是肿瘤治疗的重要靶点。HDAC抑制剂的两个主要抗癌机制是诱导终末分化和抑制细胞增殖。为了探讨HDAC在维持细胞自我更新和细胞增殖中的作用，我们用丙戊酸(VPA)和丁酸钠(NaBu)处理来自脂肪组织和脐带血的间充质干细胞(MSCs)。材料和方法：从人的乳腺脂肪组织和脐带血分离MSCs。采用四甲基偶氮唑盐比色法和流式细胞仪细胞周期分析方法评价MSCs的自我更新能力。体外分化为成骨、成脂、神经和软骨细胞系，以研究MSC的多能性。结果：VPA和NaBu使MSCs形态变平，生长受到抑制。VPA和NaBu通过增加组蛋白H3和H4的乙酰化来激活p21CIP1/WAF1的转录，最终将细胞周期阻断在G2/M期。与细胞衰老密切相关的cdk抑制剂p16INK4A的表达水平不受HDAC抑制剂的影响。我们进行了对骨、脂肪、软骨和神经组织的控制分化，以阐明HDAC在MSC分化为功能组织的多能性中所起的作用。通过特异性染色和逆转录聚合酶链式反应可以看出，VPA和NaBu降低了成脂、成软骨和成神经性分化的效率。结论：HDAC活性对于维持MSCs的自我更新和多能性是必不可少的。
Objectives:Histone deacetylase (HDAC) is an important therapeutic target in cancer. Two of the main anticancer mechanisms of HDAC inhibitors are induction of terminal differentiation and inhibition of cell proliferation. To investigate the role of HDAC in maintenance of self-renewal and cell proliferation, we treated mesenchymal stem cells (MSCs) that originated from adipose tissue or umbilical cord blood with valproic acid (VPA) and sodium butyrate (NaBu).Materials and methods:Human MSCs were isolated from mammary fat tissue and cord blood. We performed MTT assay and flow cytometry-based cell cycle analysis to assess self-renewal of MSCs. In vitro differentiation assays into osteogenic, adipogenic, neurogenic and chondrogenic lineages were conducted to investigate MSC multipotency. Immunocytochemistry, Western blot and reverse transcription-polymerase chain reaction were used to interrogate molecular pathways.Results:VPA and NaBu flattened the morphology of MSCs and inhibited their growth. VPA and NaBu activated the transcription of p21CIP1/WAF1 by increasing the acetylation of histone H3 and H4 and eventually blocked the cell cycle at G2/M phase. The expression level of p16INK4A, a cdk inhibitor that is closely related to cellular senescence, was not changed by HDAC inhibitor treatment. We performed controlled differentiation into bone, fat, cartilage and nervous tissue to elucidate the role of HDAC in the pluripotency of MSC to differentiate into functional tissues. VPA and NaBu decreased the efficiency of adipogenic, chondrogenic, and neurogenic differentiation as visualized by specific staining and reverse transcription-polymerase chain reaction. In contrast, osteogenic differentiation was elevated by HDAC inhibitor treatment.Conclusion:HDAC activity is essential for maintaining the self-renewal and pluripotency of MSCs.