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AIM:The GFAP was traditionally considered to be abiomarker for neural gila(mainly astrocytes and non-myelinating Schwann cells).Genetically,a 2.2-kb humanGFAP promoter has been successfully used to targetastrocytes in vitro and in vivo.More recently,GFAPwas also established as one of the several makers foridentifying hepatic stellate cells(HSC).In this project,possible application of the same 2.2-kb human GFAPpromoter for targeting HSC was investigated.METHODS:The GFAP-lacZ transgene was transfectedinto various cell lines(HSC,hepatocyte,and other non-HSC cell types).The transgene expression specificitywas determined by X-gal staining of the β-galactosidaseactivity.And the responsiveness of the transgene wastested with a typical pro-fibrotic cytokine TGF-β1.Theexpression of endogenous GFAP gene was assessed byreal-time RT-PCR,providing a reference for the transgeneexpression.RESULTS:The results demonstrated for the first timethat the 2.2 kb hGFAP promoter was not only capable ofdirecting HSC-specific expression,but also responding toa known pro-fibrogenic cytokine TGF-β1 by upregulationin a dose-and time-dependent manner,similar to theendogenous GFAP.CONCLUSION:In conclusion,these findings suggestednovel utilities for using the GFAP promoter to specificallymanipulate HSC for therapeutic purpose.
AIM: The GFAP was traditionally considered to be abiomarker for neural gila (mainly astrocytes and non-myelinating Schwann cells). Genetically, a 2.2-kb humanGFAP promoter has been successfully used to target astrocytes in vitro and in vivo. More recently, GFAP was also established as this one, the possible application of the same 2.2-kb human GFAP promoter for targeting HSC was investigated. METHODS: The GFAP-lacZ transgene was transfected in various cell lines (HSC, hepatocyte , and other non-HSC cell types. The transgene expression specificity is determined by X-gal staining of the β-galactosidase activity. And the responsiveness of the transgene wastested with a typical pro-fibrotic cytokine TGF-β1. The expression of endogenous GFAP gene was assessed byreal-time RT-PCR, providing a reference for the transgeneexpression.RESULTS: The results demonstrated for the first timethat the 2.2 kb hGFAP promoter was not only capable of direct ing HSC-specific expression, but also responding to known pro-fibrogenic cytokine TGF-β1 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.