TGFbeta1 regulation and collagen-release-independent connective tissue re-modelling by the ruthenium complex NAMI-A in solid tumours.
OBJECTIVE
The objective of this study is to evaluate the fibrotic process induced in vivo by NAMI-A in mice with solid tumours. In addition, the in vitro effects of NAMI-A on collagen fibres and the expression of TGFbeta1 in TS/A adenocarcinoma cells, NIH/3T3 fibroblasts and co-culture of fibroblasts and tumour cells have also been studied.


METHODS
Collagen fibres release was assayed in supernatant of culture cells treated with 0.1 and 0.01 mM NAMI-A. TGFbeta1 was detected by RT-PCR and immunoblot on cellular lysates.


RESULTS
NAMI-A, given to mice bearing MCa mammary carcinoma at advanced stages of growth, increased the thickness of connective tissue and induced recruitment of leukocytes, particularly in the peritumour capsule. In vitro NAMI-A stimulated collagen production by NIH/3T3 fibroblasts and decreased collagen release by TS/A tumour cells after prolonged exposure, either after single cell treatment or in co-cultures. In co-cultures, NAMI-A, in a dose-dependent manner, down-regulated the expression of TGFbeta1 mRNA and protein in tumour cells and up-regulated it in fibroblasts. The isoform of this cytokine is involved in fibrosis, invasion and metastatic processes.


CONCLUSIONS
These data emphasize the ability of NAMI-A to evoke beneficial effects from healthy cells against tumour growth and metastases. The contribution of fibroblasts to the fibrosis arising in tumour masses is due to TGFbeta1, and its down-regulation in tumour cells might explain the documented reduction of gelatinase release.
Callerio Foundation Onlus, via Fleming 22/31, 34127 Trieste, Italy.
Claudia
C
Casarsa
Maria Teresa
MT
Mischis
Gianni
G
Sava
D000818:Animals; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D003094:Collagen / Q000096:biosynthesis / Q000378:metabolism*; D003238:Connective Tissue / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D004121:Dimethyl Sulfoxide / Q000031:analogs & derivatives* / Q000737:chemistry* / Q000494:pharmacology*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D015674:Mammary Neoplasms, Animal / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D008808:Mice, Inbred CBA; D015394:Molecular Structure; D041681:NIH 3T3 Cells; D009942:Organometallic Compounds / Q000737:chemistry* / Q000494:pharmacology*; D012428:Ruthenium / Q000737:chemistry*; D017975:Ruthenium Compounds; D016212:Transforming Growth Factor beta / Q000096:biosynthesis / Q000235:genetics*; D053773:Transforming Growth Factor beta1
D009942:Organometallic Compounds; D017975:Ruthenium Compounds; C508961:Tgfb1 protein, mouse; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; C113661:imidazolium-bis(imidazole)dimethylsulfoxideimidazotetrachlororuthenate(III); D012428:Ruthenium; D003094:Collagen; D004121:Dimethyl Sulfoxide