PMID- 14554207
OWN - NLM
STAT- MEDLINE
DCOM- 20031118
LR  - 20211008
IS  - 0736-0266 (Print)
IS  - 0736-0266 (Linking)
VI  - 21
IP  - 6
DP  - 2003 Nov
TI  - cDNA-arrays and real-time quantitative PCR techniques in the investigation of 
      chronic Achilles tendinosis.
PG  - 970-5
AB  - The aetiology and pathogenesis of chronic painful Achilles tendinosis are 
      unknown. This investigation aimed to use cDNA arrays and real-time quantitative 
      polymerase chain reaction (real-time PCR) technique to study tendinosis and 
      control tissue samples. Five patients (females mean age 57.1+/-4.3 (years+/-SD)) 
      with chronic painful Achilles tendinosis were included. From all patients, one 
      biopsy was taken from the area with tendinosis and one from a clinically normal 
      area (control) of the tendon. The tissue samples were immediately immersed in 
      RNAlater and frozen at -80 degrees C until RNA extraction. Portions of pooled RNA 
      from control and tendinosis sites, respectively, were transcribed to cDNA, 
      radioactively labelled (32P), hybridized to cDNA expression arrays, and exposed 
      to phosphoimager screens over night. Expressions of specific genes, shown to be 
      regulated in the cDNA array analysis, were analyzed in the individual samples 
      using real-time PCR. cDNA arrays showed that gene expressions for 
      matrix-metalloproteinase-2 (MMP-2), fibronectin subunit B (FNRB), vascular 
      endothelial growth factor (VEGF), and mitogen-activated protein kinase p38 
      (MAPKp38) were up-regulated, while matrix-metalloproteinase-3 (MMP-3) and decorin 
      were down-regulated, in tendinosis tissue compared with control tissue. Using 
      real-time PCR, 4/5 and 3/5 patients showed up-regulation of MMP-2 and FNRB mRNA, 
      respectively. For decorin, VEGF, and MAPKp38, real-time PCR revealed a great 
      variability among patients. Interestingly, the mRNAs for several cytokines and 
      cytokine receptors were not regulated, indicating the absence of an inflammatory 
      process in chronic painful Achilles tendinosis. In conclusion, cDNA-arrays and 
      real-time PCR can be used to study differences in gene expression levels between 
      tendinosis and control tendon tissue.
FAU - Alfredson, Hakan
AU  - Alfredson H
AD  - Department of Surgical and Perioperative Science, Sports Medicine Unit, 
      University of Umea, S-901 87 Umea, Sweden. hakan.alfredson@idrott.umu.se
FAU - Lorentzon, Mattias
AU  - Lorentzon M
FAU - Backman, Stina
AU  - Backman S
FAU - Backman, Assar
AU  - Backman A
FAU - Lerner, Ulf H
AU  - Lerner UH
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Orthop Res
JT  - Journal of orthopaedic research : official publication of the Orthopaedic 
      Research Society
JID - 8404726
RN  - 0 (DCN protein, human)
RN  - 0 (Decorin)
RN  - 0 (Extracellular Matrix Proteins)
RN  - 0 (Fibronectins)
RN  - 0 (Proteoglycans)
RN  - 0 (RNA, Messenger)
RN  - 0 (Vascular Endothelial Growth Factor A)
RN  - 9007-49-2 (DNA)
RN  - EC 2.7.11.24 (Mitogen-Activated Protein Kinases)
RN  - EC 2.7.11.24 (p38 Mitogen-Activated Protein Kinases)
RN  - EC 3.4.24.- (Matrix Metalloproteinases)
SB  - IM
MH  - Achilles Tendon/*metabolism
MH  - DNA/analysis
MH  - Decorin
MH  - Extracellular Matrix Proteins
MH  - Female
MH  - Fibronectins/genetics/metabolism
MH  - *Gene Expression Profiling
MH  - Gene Expression Regulation
MH  - Humans
MH  - Matrix Metalloproteinases/genetics/metabolism
MH  - Middle Aged
MH  - Mitogen-Activated Protein Kinases/genetics/metabolism
MH  - Oligonucleotide Array Sequence Analysis/*methods
MH  - Proteoglycans/genetics/metabolism
MH  - RNA, Messenger/genetics
MH  - *Reverse Transcriptase Polymerase Chain Reaction
MH  - Tendinopathy/genetics/*metabolism/pathology
MH  - Vascular Endothelial Growth Factor A/genetics/metabolism
MH  - p38 Mitogen-Activated Protein Kinases
EDAT- 2003/10/14 05:00
MHDA- 2003/12/03 05:00
CRDT- 2003/10/14 05:00
PHST- 2003/10/14 05:00 [pubmed]
PHST- 2003/12/03 05:00 [medline]
PHST- 2003/10/14 05:00 [entrez]
AID - S0736026603001074 [pii]
AID - 10.1016/S0736-0266(03)00107-4 [doi]
PST - ppublish
SO  - J Orthop Res. 2003 Nov;21(6):970-5. doi: 10.1016/S0736-0266(03)00107-4.