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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(P<0.01);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(P<0.01);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(P<0.05)。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。
Thymosin beta 10 (Tβ10) gene is a predictor of tumorigenesis and grade. The study found that Tβ10 is highly expressed in the growth center of Cervidae, whereas pilose antlers can grow rapidly (2 cm / d) without any carcinogenesis. In order to investigate the effect of endogenous deer Tβ10 gene overexpression on the biological characteristics of antler stem cells. In this study, high purity AP cells were obtained by collecting the tissue of antlerogenic periosteum (AP) tissue and primary culture. The Tβ10 gene was cloned and the lentivirus overexpression vector was constructed. The lentiviral particles were packaged in vitro and infected with AP cells. The q-RT-PCR results showed that the expression of Tβ10 gene was up-regulated by 2.5-fold compared with the control, Cell line. Methyl thiazolyl tetrazolium assay (MTT) showed that overexpression of Tβ10 significantly inhibited the proliferation of AP cells (P <0.01); β-galactosidase staining showed that Tβ10 over-expression (P <0.01). The cell scratch assay showed that the overexpression of Tβ10 significantly inhibited the cell migration of human umbilical vein endothelial cells (HUVEC) P <0.05). This study provides a theoretical basis for revealing the rapid growth mechanism of antler without carcinogenesis and in-depth study of the function of deer Tβ10 gene in the rapid growth of antler.