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人胚肾293细胞产生的生物活性甲胎蛋白的纯化与特性分析

Purification and characterization of a bioactive alpha-fetoprotein produced by HEK-293 cells.

作者信息

Lin Bo, Peng Guoqing, Feng Haipeng, Li Wei, Dong Xu, Chen Yi, Lu Yan, Wang Qiaoyun, Xie Xieju, Zhu Mingyue, Li Mengsen

机构信息

Hainan Provincial Key Laboratory of Carcinogenesis and Intervention, Hainan Medical College, Haikou 571199, Hainan Province, PR China; Key Laboratory of Molecular Biology, Hainan Medical College, Haikou 571199, PR China.

Hainan Provincial Key Laboratory of Carcinogenesis and Intervention, Hainan Medical College, Haikou 571199, Hainan Province, PR China; Department of Pathophysiology, Hainan Medical College, Haikou 571199, PR China.

出版信息

Protein Expr Purif. 2017 Aug;136:1-6. doi: 10.1016/j.pep.2017.05.008. Epub 2017 May 26.

Abstract

Alpha-fetoprotein (AFP) is a biomarker that is used to diagnose hepatocellular carcinoma (HCC) and can promote malignancy in HCC. AFP is an important target in the treatment of liver cancer. To obtain enough AFP to screen for AFP inhibitors, we expressed and purified AFP in HEK-293 cells. In the present study, we produced AFP in the cells and harvested highly pure rAFP (or recombinant expression AFP in HEK-293 cells). We also analysed the bioactivity of rAFP and found that rAFP promoted growth of the human HCC cells, antagonize paclitaxel inhibition of HCC cell proliferation, suppress expression of active caspase-3, and promote expression of Ras and survivin. This study provides a method to produce significant amounts of AFP for use in biochemical assays and functional studies and to screen AFP inhibitors for use in HCC therapy.

摘要

甲胎蛋白(AFP)是一种用于诊断肝细胞癌(HCC)的生物标志物,且可促进HCC的恶性发展。AFP是肝癌治疗中的一个重要靶点。为获得足够的AFP以筛选AFP抑制剂,我们在HEK-293细胞中表达并纯化了AFP。在本研究中,我们在细胞中制备了AFP,并收获了高纯度的rAFP(或HEK-293细胞中的重组表达AFP)。我们还分析了rAFP的生物活性,发现rAFP促进人HCC细胞生长,拮抗紫杉醇对HCC细胞增殖的抑制作用,抑制活性半胱天冬酶-3的表达,并促进Ras和生存素的表达。本研究提供了一种制备大量AFP用于生化检测和功能研究的方法,并用于筛选用于HCC治疗的AFP抑制剂。

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