摘要
AIM： The GFAP was traditionally considered to be a biomarker for neural gila （mainly astrocytes and nonmyelinating 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 nonHSC cell types）. The transgene expression specificity was determined by X-gal staining of the β-galactosidase activity. And the responsiveness of the transgene was tested with a typical pro-fibrotic cytokine TGF-β1. 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-β1 by upregulation in a doseand 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.
瞄准：GFAP 传统地被认为是为神经神经胶质的一个简历标记(主要星形细胞和 non-myelinating Schwann 房间) 。遗传上，一个 2.2-kb 人 GFAP 倡导者成功地被用来指向星形细胞在试管内和在活体内。更最近， GFAP 也为识别肝的星形细胞(HSC ) 作为几个制造者之一被建立。在这个工程，为指向 HSC 的一样的 2.2-kb 人 GFAP 倡导者的可能的申请被调查。方法：GFAP-lacZ transgene 是进各种各样的房间线(HSC， hepatocyte，和另外的 non-HSC 房间类型) 的 transfected。transgene 表示特性被贝它牛乳糖活动染色的 X 女郎决定。并且 transgene 的应答的海角与典型 pro-fibrotic cytokine TGF-beta1 被测试。内长的 GFAP 基因的表示被即时 RT-PCR 估计，提供为 transgene 表示的一本参考书。结果：结果第一次证明 2.2 kb hGFAP 倡导者不仅能够指导 HSC 特定的表示，而且对已知的 pro-fibrogenic cytokine TGF-beta1 作出回应由在在一个剂量依赖者和时间依赖者举止的规定上面，类似于内长的 GFAP。结论：在结论，这些调查结果为使用 GFAP 倡导者明确地为治疗学的目的操作 HSC 建议了新奇实用程序。
基金
Supported by the Biomedical Research Council
the Institute of Bioengineering and Nanotechnology,the Republic of Singapore
作者简介
Correspondence to： Dr Lang Zhuo, Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, #04-01, 138669, Singapore. lzhuo@ibn.a-star.edu.sg Telephone： ＋65-68247114 Fax： ＋65-64789080
