Interleukin-1 beta-modulated gene expression in immortalized human chondrocytes.
Immortalized human chondrocytes were established by transfection of primary cultures of juvenile costal chondrocytes with vectors encoding simian virus 40 large T antigen and selection in suspension culture over agarose. Stable cell lines were generated that exhibited chondrocyte morphology, continuous proliferative capacity (> 80 passages) in monolayer culture in serum-containing medium, and expression of mRNAs encoding chondrocyte-specific collagens II, IX, and XI and proteoglycans in an insulin-containing serum substitute. They did not express type X collagen or versican mRNA. These cells synthesized and secreted extracellular matrix molecules that were reactive with monoclonal antibodies against type II collagen, large proteoglycan (PG-H, aggrecan), and chondroitin-4- and chondroitin-6-sulfate. Interleukin-1 beta (IL-1 beta) decreased the levels of type II collagen mRNA and increased the levels of mRNAs for collagenase, stromelysin, and immediate early genes (egr-1, c-fos, c-jun, and jun-B). These cell lines also expressed reporter gene constructs containing regulatory sequences (-577/+3,428 bp) of the type II collagen gene (COL2A1) in transient transfection experiments, and IL-1 beta suppressed this expression by 50-80%. These results show that immortalized human chondrocytes displaying cartilage-specific modulation by IL-1 beta can be used as a model for studying normal and pathological repair mechanisms.
Goldring
M B
MB
Arthritis Research Laboratory, Massachusetts General Hospital, Charlestown 02129.
Birkhead
J R
JR
Suen
L F
LF
Yamin
R
R
Mizuno
S
S
Glowacki
J
J
Arbiser
J L
JL
Apperley
J F
JF
D000957:Antigens, Viral, Tumor / Q000302:isolation & purification; D002356:Cartilage / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D002460:Cell Line / Q000187:drug effects / Q000502:physiology*; D002472:Cell Transformation, Viral; D002809:Chondroitin Sulfates / Q000302:isolation & purification; D003094:Collagen / Q000096:biosynthesis* / Q000235:genetics; D003513:Cycloheximide / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D007150:Immunohistochemistry; D007375:Interleukin-1 / Q000494:pharmacology*; D010641:Phenotype; D011509:Proteoglycans / Q000302:isolation & purification; D012333:RNA, Messenger / Q000032:analysis; D012272:Ribs / Q000166:cytology / Q000502:physiology; D013539:Simian virus 40 / Q000235:genetics
D000957:Antigens, Viral, Tumor; D007375:Interleukin-1; D011509:Proteoglycans; D012333:RNA, Messenger; D002809:Chondroitin Sulfates; D003094:Collagen; D003513:Cycloheximide