Effects of transforming growth factor-beta1 and tumour necrosis factor-alpha on cultured fibroblasts from skin fibroma as modulated by toremifene.
To determine how toremifene, an anti-oestrogen triphenylethylene derivate, reduces tumour mass, we investigated its modulation of TGF-beta1 and TNF-alpha in fibroma fibroblasts. Normal and fibroma fibroblasts, isolated from patients affected by Gardner's syndrome without or with fibroma manifestation, were cultured in vitro. Secretion of GAG, collagen and TGF-beta1 was increased in fibroma fibroblasts compared to healthy cells. The increase in TGF-beta1 secretion into the medium was associated with a parallel increase in TGF-beta1 gene expression and receptor number. Receptor cross-linking studies using radiolabelled TGF-beta1 revealed more receptors, particularly types I and II, in fibroma fibroblasts than in normal cells. Normal and fibroma fibroblasts did not synthesise TNF-alpha, but they had TNF-alpha membrane receptors, as shown by TNF-alpha assay. TNF-alpha secreted by human monocytes, which may be present in the peritumoral area, increased cell proliferation and GAG accumulation and was, in turn, enhanced by TGF-beta1 treatment. Both growth factors increased angiogenesis, as shown by the CAM assay. Toremifene reduced TGF-beta1 secretion by fibroma fibroblasts and TNF-alpha secretion by monocytes, thus downregulating cell proliferation, ECM macromolecule accumulation and angiogenic progression. We hypothesise that increased TGF-beta1 gene expression and TGF-beta1 secretion in fibroma fibroblasts as well as the subsequent rise in TNF-alpha production by monocytes may facilitate fibroma growth and that toremifene inhibits autocrine and paracrine growth factor production.
Lilli
Cinzia
C
Department of Experimental Medicine and Biochemical Science, University of Perugia, Perugia, Italy.
Marinucci
Lorella
L
Bellocchio
Silvia
S
Ribatti
Domenico
D
Balducci
Chiara
C
Baroni
Tiziano
T
Cagini
Lucio
L
Giustozzi
Giammario
G
Locci
Paola
P
D000818:Animals; D018931:Antineoplastic Agents, Hormonal / Q000627:therapeutic use*; D015152:Blotting, Northern; D002478:Cells, Cultured / Q000187:drug effects / Q000378:metabolism; D002645:Chickens; D003094:Collagen / Q000378:metabolism; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D005350:Fibroma / Q000188:drug therapy* / Q000209:etiology / Q000378:metabolism; D005736:Gardner Syndrome / Q000150:complications; D006025:Glycosaminoglycans / Q000378:metabolism; D006801:Humans; D012333:RNA, Messenger / Q000096:biosynthesis; D012334:RNA, Neoplasm / Q000096:biosynthesis; D011960:Receptors, Estrogen / Q000378:metabolism; D018125:Receptors, Transforming Growth Factor beta / Q000378:metabolism; D012878:Skin Neoplasms / Q000188:drug therapy* / Q000209:etiology / Q000378:metabolism; D017312:Toremifene / Q000627:therapeutic use*; D016212:Transforming Growth Factor beta / Q000378:metabolism* / Q000494:pharmacology; D053773:Transforming Growth Factor beta1; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism* / Q000494:pharmacology
D018931:Antineoplastic Agents, Hormonal; D006025:Glycosaminoglycans; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D011960:Receptors, Estrogen; D018125:Receptors, Transforming Growth Factor beta; C508960:TGFB1 protein, human; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D014409:Tumor Necrosis Factor-alpha; D017312:Toremifene; D003094:Collagen