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Cloning and promoter activity of rat Smad1 5′-flanking region in rat hepatic stellate cells

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Abstract

Smad1 is an important signaling molecule for members of transforming growth factor-β (TGF-β) superfamily. Increased expression of Smad1 in activated hepatic stellate cells (HSCs) indicates a role of Smad1 in liver fibrosis. Therefore, understanding of Smad1 gene expression could be important to control the activation of HSCs. Current study reports the cloning and characterizing rat Smad1 5′-flanking region in liver cells. Rat Smad1 5′-flanking region was cloned by PCR method. Promoter deletional analysis and electrophoretic mobility shift assay (EMSA) were examined in hepatocyte and HSCs cell line (CFSC-8B cells), respectively. Results indicated that rat Smad1 used GC-box as its promoter and there was a transcriptional regulatory element located at the region of −163 to −56bp. EMSA demonstrated two bands on Smad1 promoter region. Smad1 promoter activity was higher in CFSC-8B cells cultured on uncoated plastic dish than that of CFSC-8B cells cultured on Matrigel-coated plastic dish. In conclusion, rat Smad1 promoter was cloned and characterized in hepatocyte and HSC cell line (CFSC-8B cells) at different culture conditions.

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Acknowledgement

This work was supported by the grant (MOP-62923) from Canadian Institute of Health Research to Dr. Yuewen Gong.

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Authors and Affiliations

  1. Faculty of Pharmacy, University of Manitoba, 50 Sifton Road, Winnipeg, MB, Canada, R3T 2N2

    Yu Sun, Jianghong Fan, Hong Shen & Yuewen Gong

  2. Department of Pharmacology, China–Japan Friendship Hospital, Beijing, China

    Ping Li

  3. Department of Physiology, Faculty of Medicine, University of Manitoba, Winnipeg, MB, Canada

    Peter Cattini

  4. Section of Hepatology of the Department of Internal Medicine, Faculty of Medicine, University of Manitoba, Winnipeg, MB, Canada

    Yuewen Gong

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  1. Yu Sun
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  2. Jianghong Fan
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  3. Hong Shen
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  4. Ping Li
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  5. Peter Cattini
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Correspondence to Yuewen Gong.

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Sun, Y., Fan, J., Shen, H. et al. Cloning and promoter activity of rat Smad1 5′-flanking region in rat hepatic stellate cells. Mol Cell Biochem 304, 227–234 (2007). https://doi.org/10.1007/s11010-007-9504-8

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  • DOI: https://doi.org/10.1007/s11010-007-9504-8

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