The inhibitory effects of camptothecin, a topoisomerase I inhibitor, on collagen synthesis in fibroblasts from patients with systemic sclerosis

Arthritis Res. 2001;3(5):311-8. doi: 10.1186/ar321. Epub 2001 Aug 2.

Abstract

The main manifestation of systemic sclerosis (SSc) is the overproduction of extracellular matrix, predominantly type I collagen. This study was undertaken to evaluate the effects of noncytotoxic doses of the topoisomerase I inhibitor camptothecin (CPT) on collagen production in the activated dermal fibroblasts from patients with SSc and healthy donors. The fibroblasts were cultured in the presence or absence of CPT. Production of collagenous proteins by fibroblasts was determined in cell and matrix layers by ELISA and in conditioned media by [(3)H]proline incorporation, gel electrophoresis, and autoradiography. Expression of alpha2(I) collagen (COL1A2) mRNA was measured by northern blot, and the activity of COL1A2 promoter was determined by a chloramphenicol acetyltransferase assay. CPT (10(-7) M) decreased the deposition of type I collagen by 68%, of type III by 38%, and of type VI by 21% in SSc fibroblasts and to a lesser degree in healthy controls. Similarly, CPT (10(-8) M to 10(-6) M) significantly inhibited secretion of newly synthesized collagenous proteins into conditioned media by 50%. CPT (10(-8) M to 10(-6) M) caused a significant dose-dependent inhibition of COL1A2 mRNA levels and COL1A2 promoter activity, both by as much as 60%. The inhibitory effect of CPT on collagen production by fibroblasts from patients with SSc suggests that topoisomerase I inhibitors may be effective in limiting fibrosis in such patients.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adult
  • Camptothecin / pharmacokinetics*
  • Cells, Cultured
  • Collagen / biosynthesis*
  • Collagen / drug effects
  • Collagen / genetics
  • Culture Media, Conditioned / chemistry
  • Dose-Response Relationship, Drug
  • Elastin / biosynthesis
  • Elastin / drug effects
  • Elastin / genetics
  • Enzyme Inhibitors / pharmacology*
  • Enzyme-Linked Immunosorbent Assay
  • Extracellular Matrix / chemistry
  • Extracellular Matrix / drug effects
  • Extracellular Matrix / metabolism
  • Female
  • Fibroblasts / drug effects*
  • Fibroblasts / metabolism
  • Humans
  • Male
  • Procollagen / biosynthesis
  • Procollagen / drug effects
  • Procollagen / genetics
  • RNA, Messenger / metabolism
  • Scleroderma, Systemic / metabolism*
  • Skin / cytology
  • Skin / drug effects
  • Skin / metabolism
  • Topoisomerase I Inhibitors*

Substances

  • Culture Media, Conditioned
  • Enzyme Inhibitors
  • Procollagen
  • RNA, Messenger
  • Topoisomerase I Inhibitors
  • Collagen
  • Elastin
  • Camptothecin