근육세포 분화에 대한 TGF-β1과 OP-1의 억제 효과
The Inhibitory Effect of TGF-β1 and OP-1 onto the Myogenic Differentiation
김병국, 정성수,
소속 상세정보
김병국 ( ) - 전북대학교 의과대학 간호학과
정성수 ( ) - 전남대학교 치과대학 마취과학
KMID : 0355220010260010039
Abstract
In order to investigate the effect of Transforming growth factor β1(below TGF-β1) and osteogenic protein-1(below OP-1) onto the myogenic differentiation, C2C12 statellite myoblastic cell line was cultured and treated with both growth factors. At first morphological changes with microscopical examination were examined, and isolated total RNA to analyse mRNA expression of bone marker proteins, muscle regulatory proteins, TGF-β receptor and their ligands by Northen blot analysis. And cellular proliferative inducibility of both growth factors was also tested to C2C12 cells.
Incubating the cell with 5 ng/㎖ of TGF-β1 until 4 days almost inhibited multinucleated mytoube formation expressing muscular regulatory proteins, and induced decreasing Id proteins. However, no osteoblastic phenotypes was induced by TGF-β1 in C2C12 cells. The mRNA expression of TGF-β receptors with TGF-β1 was conversed after 48 hours cultured. Type ⅠTGF-β receptor was seemed to play a role in negative signalling for inhibition of myogenic differentiation. OP-1 dose dependently induced ALP activity, osteopontine production and bone sialoprotein production at concentration above 100ng/㎖ and osteocalcin production at concentrations above 300ng/㎖. The concentration of OP-1 required to induce these osteoblastic phenotypes was the same as that required to almost completely inhibit myotube formation. Incubation with above 100ng/㎖ OP-1 suppressed the expression of mRNA for muscular regulatory proteins from 2 days after incubation. Expression of Id-1, 2, 3 mRNA were stimulated by OP-1 at concentration above 300ng/㎖. When C2C12 cells were treated with both growth factors, TGF-β1 potentiated the inhibitory effect of OP-1 on myotube formation and expression of mRNA for myogenin at 12 days. And TGF-β1 reduced osteocalcin and bone sialoprotein production induced by OP-1 at 12 days in C2C12 cells. Both growth factor had no mitogenic effect. These results indicate that OP-1 converts the differentiation pathway of C2C12 myoblasts into that of osteoblastic lineage cells and it´s not heritable, but TGF-β1 dose not and has reversible inhibitory activity on the myogenic differentiation. TGF-β1 and OP-1 play a role in myogenic differentiation via different mechanism between them.
키워드
원문 및 링크아웃 정보
등재저널 정보