人肝脏再生增强因子对体外肝细胞生长的影响

李曼妮; 苏先狮

人肝脏再生增强因子对体外肝细胞生长的影响

1. 广东省公安边防总队医院二内科
2. 中南大学湘雅二医院肝病研究中心

详细信息

中图分类号: R575.29
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出版历程
- 出版日期: 2009-05-20

摘要

摘要: 目的基因工程方法纯化人肝脏再生增强因子（hALR）,研究hALR对体外培养肝细胞生长的影响。方法构建hALR原核表达载体PGEX-3X-hALR,诱导表达、纯化收集GST-hALR,用X因子切去GST,凝胶过滤得到高纯度hALR蛋白;分别用正常肝细胞（L-02细胞）和肝癌细胞（HepG2细胞）分析hALR对其生长的影响。结果hALR原核表达载体pGEX-3X-hALR构建成功,经诱导表达,可纯化得到高纯度的hALR蛋白。hALR蛋白能促进正常肝细胞的生长,并与浓度成正相关;而肝癌细胞的生长却起抑制作用。结论成功表达和纯化重组hALR蛋白,hALR促进正常肝细胞的生长而抑制肝癌细胞生长。
- 肝再生增强因子:克隆表达:肝细胞生长
Abstract: Objective To purify the human augmenter of liver regeneration (hALR) by gene engineering and to research the influence on the growth of liver cell cultured in vitro.Methods The hALR cDNA was obtained by using RT-PCR, then was cloned into the PGEX-3X and transformed into E coli.The fusion protein GST-hALR was produced by IPTG.The hALR protein was purified after cutted in vitro, and then was anlyzed.Results The recombinant expression plasmid PGEX-3X-hALR was proved to be corrected by sequencing.The molecular weight of expressed product was 41 KD as expected as GST-hALR molecular weight in SDS-PAGE.The recombinant hALR has effects on prompting the liver cell regeneration and inhibiting the hepatoma carcinoma cell regeneration.Conclusion The recombinant hALR protein was obtained and purified successfully.And it has effects on prompting the liver cell regeneration and inhibiting the hepatoma carcinima cell regeneration.
- augmenter of liver regeneration

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参考文献(9)

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