胶质纤维酸性蛋白（GFAP）启动子特异性地指导大鼠肝星状细胞系中的β-半乳糖苷酶（lacZ）表达。
GFAP promoter directs lacZ expression specifically in a rat hepatic stellate cell line.
作者信息
Maubach Gunter, Lim Michelle Chin Chia, Zhang Chun-Yan, Zhuo Lang
机构信息
Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, #04-01, 138669, Singapore.
出版信息
World J Gastroenterol. 2006 Feb 7;12(5):723-30. doi: 10.3748/wjg.v12.i5.723.
AIM
The GFAP was traditionally considered to be a biomarker for neural glia (mainly astrocytes and non-myelinating Schwann cells). Genetically, a 2.2-kb human GFAP promoter has been successfully used to target astrocytes in vitro and in vivo. More recently, GFAP was also established as one of the several makers for identifying hepatic stellate cells (HSC). In this project, possible application of the same 2.2-kb human GFAP promoter for targeting HSC was investigated.
METHODS
The GFAP-lacZ transgene was transfected into various cell lines (HSC, hepatocyte, and other non-HSC cell types). The transgene expression specificity was determined by X-gal staining of the beta-galactosidase activity. And the responsiveness of the transgene was tested with a typical pro-fibrotic cytokine TGF-beta1. The expression of endogenous GFAP gene was assessed by real-time RT-PCR, providing a reference for the transgene expression.
RESULTS
The results demonstrated for the first time that the 2.2 kb hGFAP promoter was not only capable of directing HSC-specific expression, but also responding to a known pro-fibrogenic cytokine TGF-beta1 by upregulation in a dose- and time-dependent manner, similar to the endogenous GFAP.
CONCLUSION
In conclusion, these findings suggested novel utilities for using the GFAP promoter to specifically manipulate HSC for therapeutic purpose.
目的
传统上认为胶质纤维酸性蛋白（GFAP）是神经胶质细胞（主要是星形胶质细胞和无髓鞘雪旺细胞）的生物标志物。在基因层面，一个2.2 kb的人类GFAP启动子已成功用于在体外和体内靶向星形胶质细胞。最近，GFAP也被确立为鉴定肝星状细胞（HSC）的几种标志物之一。在本项目中，研究了相同的2.2 kb人类GFAP启动子靶向HSC的可能应用。
方法
将GFAP-lacZ转基因转染到各种细胞系（HSC、肝细胞和其他非HSC细胞类型）中。通过β-半乳糖苷酶活性的X-gal染色确定转基因表达特异性。并用典型的促纤维化细胞因子TGF-β1测试转基因的反应性。通过实时RT-PCR评估内源性GFAP基因的表达，为转基因表达提供参考。
结果
结果首次表明，2.2 kb的人类GFAP启动子不仅能够指导HSC特异性表达，而且能够以剂量和时间依赖性方式对已知的促纤维化细胞因子TGF-β1作出反应，通过上调表达，类似于内源性GFAP。
结论
总之，这些发现提示了利用GFAP启动子特异性操纵HSC用于治疗目的的新用途。
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