Collagen biosynthesis in human oral submucous fibrosis fibroblast cultures.
To investigate the mechanism of collagen accumulation in oral submucous fibrosis (OSF) tissues, we examined the biosynthesis of collagen in fibroblast cultures established from OSF lesions. Fibroblasts obtained from four of ten OSF specimens showed more than a 1.5-fold increase in the production of collagens compared with fibroblasts from age-, sex-, and passage-matched normal controls (p < 0.05). When the relative amounts of collagen synthesis were estimated by SDS polyacrylamide gel electrophoresis, it was found that both OSF and control cells produced about 85% type I collagen and 15% type III collagen. The ratio of alpha 1(I) to alpha 2(I) chains was about 3:1 in OSF cells instead of the 2:1 expected for type I collagen. The excess alpha 1(I) chains could mean that collagen type I trimer was synthesized by the fibroblasts. These findings suggest that collagen overproduction and a reduced degradation of the structure-stable collagen type I trimer synthesized by OSF fibroblasts might contribute to the accumulation of collagen in OSF lesions in vivo. The mechanism(s) of increased procollagen production were analyzed by Northern blot, slot blot, and Southern blot. The OSF fibroblast strains with elevated collagen production also contained higher-than-normal levels of procollagen mRNA, and the ratios of alpha 1(I), alpha 2(I), and alpha 1(III) procollagen mRNAs were compatible with the results of corresponding procollagen alpha chains. The gene copy number of pro alpha 2(I) collagen gene in OSF fibroblasts was about 1.05. No gene amplification was found. These results indicate that expression of these procollagen genes in cultured fibroblasts is regulated at the transcriptional level.
Kuo
M Y
MY
School of Dentistry, College of Medicine, National Taiwan University, Taipei.
Chen
H M
HM
Hahn
L J
LJ
Hsieh
C C
CC
Chiang
C P
CP
D016022:Case-Control Studies; D002478:Cells, Cultured / Q000378:metabolism; D003094:Collagen / Q000096:biosynthesis*; D005347:Fibroblasts / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D015151:Immunoblotting; D008297:Male; D009061:Mouth Mucosa / Q000378:metabolism; D009914:Oral Submucous Fibrosis / Q000235:genetics / Q000378:metabolism*; D011347:Procollagen / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis
D011347:Procollagen; D012333:RNA, Messenger; D003094:Collagen