A feasible tool to detect mRNA expression of matrix metalloproteinases and their tissue inhibitors in human Tenon's capsule.
OBJECTIVE
To determine the mRNA expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in human specimens of Tenon's capsule.


METHODS
Reverse transcription-polymerase chain reaction (RT-PCR) with specific primers for MMP-1, MMP-2, MMP-9, TIMP-1 and TIMP-2 was performed on tissue specimens obtained from patients with cataract, rhegmatogenous retinal detachment, glaucoma, or diabetes mellitus.


RESULTS
Glyceraldehyde phosphate dehydrogenase (GAPDH) mRNA transcripts were detected in 26 (76.5%) of 34 specimens, with almost the same amount of expression in each of these samples. Messenger RNA expression of one or more of the MMPs/TIMPs could also be detected in all of these 26 samples, but not in any sample without GAPDH expression. MMP-2, TIMP-1 and TIMP-2 were detected in 25 (96.2%) of the 26 samples with GAPDH expression, while MMP-1 and MMP-9 were detected with a lower percentage (34.6 and 19.2%, respectively).


CONCLUSIONS
The feasibility of RT- PCR with GAPDH as an internal standard to determine mRNA transcripts of the MMPs and TIMPs in the subconjunctival Tenon's capsule was demonstrated.
Huang
Yu-Lun
YL
Department of Medical Research, Chi-Mei Foundation Medical Center, Taipei, Taiwan.
Liu
Catherine J
CJ
Chiu
Allen W
AW
Wang
Yun-Chun
YC
Huan
Steven K
SK
Lee
Fenq-Lih
FL
Chen
Shih-Jen
SJ
Hsu
Wen-Ming
WM
Hsieh
Shie-Liang
SL
D003228:Conjunctiva / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D005123:Eye / Q000166:cytology* / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D020782:Matrix Metalloproteinases / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D019714:Tissue Inhibitor of Metalloproteinases / Q000235:genetics / Q000378:metabolism*
D017931:DNA Primers; D012333:RNA, Messenger; D019714:Tissue Inhibitor of Metalloproteinases; D020782:Matrix Metalloproteinases