Effects of the extracellular matrix on lumican expression in rat aortic smooth muscle cells in vitro.
Lumican is a small leucine-rich proteoglycan (SLRP), which contributes to cell migration, proliferation, tissue hydration, and collagen fibrillogenesis. Whether lumican is localized in rat aortic smooth muscle cells (SMCs) and what its relationships might be to other extracellular matrix components have not yet been elucidated. In this study, using reverse transcription-polymerase chain reaction (RT-PCR), competitive RT-PCR, and western blot, lumican messenger ribonucleic acid (mRNA) was expressed in cultured rat aortic SMCs. SMCs cultured in serum-free medium showed four bands at 68, 62, 50, and 37 kD. The 68 and 62 kD bands corresponded to proteoglycan, the 50 kD band to glycoprotein, and the 37 kD band to the core protein form of lumican. The relationships of lumican to fibronectin and laminin were also investigated. The lumican mRNA level in SMCs cultured on fibronectin was highest at day 1, but it increased at day 3 in SMCs cultured on laminin. On the fibronectin or laminin-coated plates, SMCs expressed only the 68 and 62 kD bands, corresponding to proteoglycan. Pretreatment with anti-beta1 integrin receptor antibody revealed a decrease in the proteoglycan forms of lumican protein and an additional two bands at 50 and 37 kD, indicating glycoprotein and the core protein of lumican. These results show that lumican was synthesized in cultured rat aortic SMCs as proteoglycan, glycoprotein, and core protein. The extracellular matrix (ECM) affected lumican protein production and restricted the lumican protein form to proteoglycan via the beta1 integrin receptor in SMCs.
Qin
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Department of Pathology, Nippon Medical School, Tokyo 113-8602, Japan.
Ishiwata
T
T
Asano
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G
D000818:Animals; D001013:Aorta, Thoracic / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D015153:Blotting, Western; D018929:Cell Culture Techniques; D011508:Chondroitin Sulfate Proteoglycans / Q000096:biosynthesis* / Q000235:genetics; D016326:Extracellular Matrix Proteins / Q000494:pharmacology*; D005353:Fibronectins / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D019012:Integrin beta1 / Q000502:physiology; D007632:Keratan Sulfate / Q000096:biosynthesis* / Q000235:genetics; D007797:Laminin / Q000494:pharmacology; D000071234:Lumican; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats
D011508:Chondroitin Sulfate Proteoglycans; D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D019012:Integrin beta1; D007797:Laminin; C000604308:Lum protein, rat; D000071234:Lumican; D012333:RNA, Messenger; D007632:Keratan Sulfate