Expression profile of protein tyrosine kinase genes in human osteoarthritis chondrocytes.
OBJECTIVE
To determine the expression profile of protein kinase (PK) and protein tyrosine kinase (PTK) genes in human primary osteoarthritis (OA) chondrocytes and to compare it with that of immortalized human chondrocytes T/C 28a4 with a view to learning whether T/C 28a4 cells can be used for elucidating signal transduction pathways in human chondrocytes.


METHODS
We used the Atlas Human cDNA Array and a method based on PCR with degenerate primers to analyse the expression profile of protein kinase genes in primary human OA chondrocytes and compared it with that of immortalized human chondrocyte cell line T/C 28a4 using RT-PCR and Western blotting.


RESULTS
A total of 21 PTK genes were identified and several of these have never been shown to be expressed in human OA chondrocytes. Comparative expression analysis of some selected kinase genes showed that the mRNA expression pattern of many protein kinase genes in OA chondrocytes was identical to that of T/C 28a4 cells. However, there were differences in the level of protein expression of selected protein kinases in these cells. For example, mRNA expression of the novel kinase HCK was detected in OA chondrocytes and in the cell lines analysed but by Western blotting HCK protein was not detected in OA chondrocytes. In these studies, we also identified a novel mutant form of the discoidin domain receptor 2 (DDR2) transcript from chondrocyte-like cell line HTB-94.


CONCLUSIONS
Our results provide novel information about protein kinase gene expression in OA chondrocytes and indicate that the transformed chondrocyte cell line T/C 28a4 may be suitable for elucidating signal transduction pathways in chondrocytes and to investigate how they regulate chondrocyte function in inflammatory and degenerative joint diseases.
Islam
S
S
Department of Medicine, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, Ohio 44106, USA.
Kermode
T
T
Sultana
D
D
Moskowitz
R W
RW
Mukhtar
H
H
Malemud
C J
CJ
Goldberg
V M
VM
Haqqi
T M
TM
D015153:Blotting, Western; D002461:Cell Line, Transformed; D019902:Chondrocytes / Q000502:physiology*; D018076:DNA, Complementary; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D020869:Gene Expression Profiling; D006801:Humans; D015345:Oligonucleotide Probes; D010003:Osteoarthritis / Q000235:genetics* / Q000473:pathology; D011494:Protein Kinases / Q000235:genetics; D011505:Protein-Tyrosine Kinases / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000502:physiology
D018076:DNA, Complementary; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D011494:Protein Kinases; D011505:Protein-Tyrosine Kinases