Endothelin-1 (ET-1) promotes MMP-2 and MMP-9 induction involving the transcription factor NF-kappaB in human osteosarcoma.
In the present study, we have investigated the effect of (i) ET-1 (endothelin-1) and its precursor, big ET-1, on MMP (matrix metalloproteinase)-2 and MMP-9 synthesis and activity in osteosarcoma tissue, and (ii) ET-1 receptor antagonists on cell invasion. Using Western blotting, zymography, RT-PCR (reverse transcription-PCR), immunohistochemistry, immunofluorescence and Northern blotting, we have shown that ET-1 and ET-1 receptors (ET(A) and ET(B)) were expressed in these cells. Additionally, we have demonstrated that ET-1 markedly induced the synthesis and activity of MMP-2, which was significantly increased when compared with MMP-9. Furthermore, inhibition of NF-kappaB (nuclear factor kappaB) activation blocked MMP-2 production and activity, indicating the involvement of NF-kappaB, a ubiquitous transcription factor playing a central role in the differentiation, proliferation and malignant transformation. Since ET-1 acts as an autocrine mediator through gelatinase induction and because inhibition of ET(A) receptor is beneficial for reducing both basal and ET-1-induced osteosarcoma cell invasion, targeting this receptor could be an attractive therapeutic alternative for the successful treatment of osteosarcoma.
Felx
Mélanie
M
Research Centre of the CHU Sainte-Justine Hospital, Montreal, QC, Canada.
Guyot
Marie-Claude
MC
Isler
Marc
M
Turcotte
Robert E
RE
Doyon
Josée
J
Khatib
Abdel-Majid
AM
Leclerc
Séverine
S
Moreau
Alain
A
Moldovan
Florina
F
D000293:Adolescent; D000328:Adult; D000975:Antioxidants / Q000494:pharmacology; D015152:Blotting, Northern / Q000379:methods; D015153:Blotting, Western / Q000379:methods; D001859:Bone Neoplasms / Q000378:metabolism*; D045744:Cell Line, Tumor; D002648:Child; D019332:Endothelin-1 / Q000494:pharmacology*; D004789:Enzyme Activation; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D020778:Matrix Metalloproteinase 2 / Q000032:analysis / Q000378:metabolism*; D020780:Matrix Metalloproteinase 9 / Q000032:analysis / Q000378:metabolism*; D008875:Middle Aged; D016328:NF-kappa B / Q000032:analysis / Q000378:metabolism*; D012516:Osteosarcoma / Q000378:metabolism*; D011759:Pyrrolidines / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018709:Statistics, Nonparametric; D013859:Thiocarbamates / Q000494:pharmacology; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000032:analysis / Q000378:metabolism; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000032:analysis / Q000378:metabolism; D014407:Tumor Cells, Cultured
D000975:Antioxidants; D019332:Endothelin-1; D016328:NF-kappa B; D011759:Pyrrolidines; D013859:Thiocarbamates; D019715:Tissue Inhibitor of Metalloproteinase-1; D019716:Tissue Inhibitor of Metalloproteinase-2; C020972:pyrrolidine dithiocarbamic acid; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9