胸腺素β4真核表达载体的构建及其对血管内皮细胞增殖的影响

Construction of Thymosin Beta 4 Eukaryotic Expression Plasmid and Its Effects on Vascular Endothelial Cells

目的:构建人胸腺素β4基因的真核表达载体,初步探讨胸腺素β4对人脐静脉内皮细胞增殖的影响,以进一步研究其在肿瘤侵袭转移中的作用机制。方法:用DNA重组技术构建真核表达载体PsecTag2B/Tβ4,并采用脂质体转染技术瞬时转染SW480细胞,采用Western blot法检测胸腺素β4重组蛋白的表达,分别用四甲基偶氮唑蓝(MTT)比色法和细胞计数法,分析SW480细胞瞬时转染培养上清液对HUVECs增殖的影响。结果:成功构建了胸腺素β4真核表达载体PsecTag2B/Tβ4,并在SW480细胞中瞬时转染,Western blot法检测到胸腺素β4蛋白表达,MTT法和细胞计数法检测胸腺素β4转染后的SW480细胞培养上清液能够促进HUVECs增殖。结论:胸腺素β4能显著促进人血管内皮细胞增殖,在肿瘤血管生成中起着一定的作用。

Objective： To construct a eukaryotic expression vector encoding the gene of thymosin beta 4 （Tβ4）, and investigate its effect on the proliferation of human umbilical vein endothelial cells （HUVECs） for further reseach its role on invasion and metastasis of cancer. Methods： PsecTag2B/Tβ4 expression vector was constructed by DNA recombinant technique and was introduced into SW480 cells by lipofectactin-mediated DNA transfection. Western blot assay were used to determine Tβ4 protein expression. Effect on cell proliferation of HUVECs was tested by MTT assay and cell counting. Results： There is expression of Tβ4 protein in the SW480 cells transfected with PsecTag2B/Tβ4. The conditioned medium after transfection showed the biological activity which could stimulate the proliferation of HUVECs. Conclusion： Thymosin beta 4 could stimulate the proliferation of vascular endothelial cells in vitro and has effects on angiogenesis of carcinoma.