Tβ10基因过表达对离体鹿茸干细胞增殖、衰老和迁移的影响

发布时间：2018-04-13 14:26

本文选题：鹿 + 胸腺素β(Tβ)　；参考：《农业生物技术学报》2017年01期

【摘要】：胸腺素β10(thymosin beta 10,Tβ10)基因是肿瘤发生以及恶化分级的一个预判指标。研究发现,鹿(Cervidae)茸顶端的生长中心组织中Tβ10高度表达,而鹿茸却能快速生长(2 cm/d)不发生任何癌变。为了研究内源性鹿Tβ10基因过表达对鹿茸干细胞部分生物学特性的影响。本研究通过采集鹿生茸区骨膜(antlerogenic periosteum,AP)组织并对其进行了原代培养获得了高纯度AP细胞。通过克隆鹿Tβ10基因并构建慢病毒过表达载体,离体包装并收集慢病毒颗粒后感染AP细胞,q RT-PCR检测表明Tβ10基因表达量相对于对照上调了2.5倍,进而建立Tβ10过表达AP细胞系。四甲基偶氮唑盐比色法(methyl thiazolyl tetrazolium assay,MTT)实验表明,Tβ10基因过表达能极显著抑制AP细胞增殖(P0.01);β-半乳糖苷酶染色显示,Tβ10基因过表达极显著地促进了AP细胞的衰老(P0.01);细胞划痕实验表明,Tβ10基因过表达显著抑制人(Homo sapiens)脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)的细胞迁移(P0.05)。本研究为揭示鹿茸快速生长机制而不发生癌变以及深入研究鹿Tβ10基因在鹿茸快速生长中的功能提供了理论依据。
[Abstract]:Thymosin 尾 10(thymosin beta 10 T 尾 10) gene is a predictor of tumor progression and progression.It was found that T 尾 10 was highly expressed in the growth center of Cervidae antler tip, while the antler could grow at a rate of 2 cm / d) without any carcinogenesis.To study the effect of overexpression of endogenous deer T 尾 10 gene on some biological characteristics of pilose antler stem cells.In this study, high purity AP cells were obtained by collecting and primary culture the tissue of antlerogenic periosteum.By cloning deer T 尾 10 gene and constructing lentivirus overexpression vector, the Q RT-PCR analysis of AP cells infected with lentivirus particles in vitro showed that T 尾 10 gene expression increased 2.5 times compared with control, and then T 尾 10 overexpression AP cell line was established.The results of methyl thiazolyl tetrazolium assay-MTT assay showed that the overexpression of T 尾 10 gene could significantly inhibit the proliferation of AP cells (P 0.01); 尾 -galactosidase staining showed that the overexpression of T 尾 10 gene significantly promoted the aging of AP cells (P 0.01);The results showed that the overexpression of T 尾 10 gene significantly inhibited the migration of human umbilical vein endothelial cells (HUVECs) from human umbilical vein endothelial cells.This study provides a theoretical basis for revealing the mechanism of rapid growth of velvet antler without carcinogenesis and for further studying the function of deer T 尾 10 gene in rapid growth of velvet antler.
【作者单位】：中国农业科学院特产研究所/特种经济动物分子生物学国家重点实验/吉林省鹿茸工程研究中心;
【基金】：国家自然科学基金项目(No.31500792) 吉林省科技发展计划项目(No.20140101139JC) 中国农业科学院科技创新工程项目(No.CAAS-ASTIP-2014-ISAPS-04)
【分类号】：S825

【相似文献】