Regulatory expression of genes related to metastasis by TGF-beta and activin A in B16 murine melanoma cells.
TGF-beta induces epithelial-mesenchymal transition, which occurs during tumor cell invasiveness in pathological state, in limited cells. As a first step to understand the role of TGF-beta and the structurally related activin during melanoma metastasis, expression of metastasis-related genes was examined in murine melanoma cells. Treatment with TGF-beta1 or activin A down-regulated E-cadherin in B16 cells in a dose-dependent manner. In epithelial cells, TGF-beta-induced high mobility group A2 (HMGA2) gene product is suggested to down-regulate E-cadherin through up-regulation of zinc-finger transcription factors Slug and Snail, and basic helix-loop-helix transcription factor Twist. Unlike the regulation in epithelial cells, TGF-beta1 treatment rather decreased mRNA expression of HMGA2, indicating a distinct mechanism on TGF-beta/activin-induced down-regulation. Transfection of double-stranded interfering RNA (dsRNAi) for activin receptor-like kinase (ALK) type I receptors revealed that ALK5, a prototype of TGF-beta receptor, mainly transmits TGF-beta signals on the E-cadherin down-regulation at the mRNA level, and that a prototype receptor ALK4 elicited the activin effect. TGF-beta/activin potentiated down-regulation of E-cadherin and HMGA2 also in B16 sublines that are susceptible to metastasis. However, the extent of down-regulation tended to be smaller, and less Smad2, a signal mediator for TGF-beta/activin, was phosphorylated in response to the ligand, resulting from less expression of type I receptors in the B16 sublines. These results suggest that the receptor expression level determines strength of the signals for TGF-beta/activin through phosphorylation of Smad2, which explains pluripotency of the ligand family partly.
Murakami
Masaru
M
Laboratory of Molecular Biology, Azabu University School of Veterinary Medicine, Sagamihara, Japan.
Suzuki
Makiko
M
Nishino
Yoshii
Y
Funaba
Masayuki
M
D030201:Activin Receptors, Type I / Q000378:metabolism; D028341:Activins / Q000494:pharmacology*; D000818:Animals; D015153:Blotting, Western; D015820:Cadherins / Q000378:metabolism; D045744:Cell Line, Tumor; D017931:DNA Primers / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects* / Q000235:genetics; D008546:Melanoma, Experimental / Q000378:metabolism*; D051379:Mice; D009841:Oligonucleotides / Q000235:genetics; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000235:genetics; D000077293:Receptor, Transforming Growth Factor-beta Type I; D018125:Receptors, Transforming Growth Factor beta / Q000378:metabolism; D051899:Smad2 Protein / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000494:pharmacology*
D015820:Cadherins; D017931:DNA Primers; D009841:Oligonucleotides; D012330:RNA, Double-Stranded; D018125:Receptors, Transforming Growth Factor beta; D051899:Smad2 Protein; C497783:Smad2 protein, mouse; D016212:Transforming Growth Factor beta; C422405:activin A; D028341:Activins; D017346:Protein Serine-Threonine Kinases; D030201:Activin Receptors, Type I; C498933:Acvr1b protein, mouse; D000077293:Receptor, Transforming Growth Factor-beta Type I; C000625842:Tgfbr1 protein, mouse