- Paper Report
- Open Access
A key enzyme upregulating collagen production by scleroderma fibroblasts
- Hideto Kameda Tsutomu Takeuchi1
© Biomed Central Ltd 2002
- Received: 16 January 2002
- Published: 16 January 2002
- dermal fibroblasts
- systemic sclerosis
- transforming growth factor ?
Systemic sclerosis (SSc) is a fibroproliferative autoimmune disease characterized by the excessive production and deposition of collagen in skin and internal organs. Acting through specific transmembrane receptors, transforming growth factor-? (TGF-?) is known to play a crucial role in the development of tissue fibrosis. The authors had previously identified several signaling molecules other than Smad proteins including protein kinase C-d (PKC-d) as candidates downstream of TGF-? receptors. The aim of the present study was to examine the possible role of PKC-d in the upregulation collagen gene expression in SSc dermal fibroblasts.
Compared to normal fibroblasts, SSc dermal fibroblasts had increased type I and III collagen mRNA and protein as well as upregulation of PKC-d protein. Rottlerin, a specific inhibitor of PKC-d, inhibited collagen gene expression in both normal and SSc fibroblasts. Moreover, this study identified a 129-bp promoter region of the type I collagen gene encompassing nucleotides -804 to -675 which was responsive to the transcriptional inhibition by rottlerin and dominant-negative PKC-d expression.
ELISA, collagenase digestion assay, northern blotting, in vitro nuclear transcription assay, chloramphenicol acetyl transferase gene assay, high-resolution fluorescence immunomicroscopy, western blotting