Repression of integrin beta 1 subunit expression by antisense RNA.
A quail cell line (QT6-c) was co-transfected with pTEX vector expressing RNA complementary to chicken integrin beta 1 subunit mRNA (Anti-Int) and pRSVneo vector by a calcium phosphate method. Transfectants showing reduced expression of quail integrin beta 1 subunit were selected with an immunoblot assay, and a few positive clones were examined in detail. Northern blot and immunoblot analyses revealed that the Anti-Int caused a clear reduction of the transcript encoding integrin beta 1 subunit depending on culture conditions. The number of cell surface integrins also decreased in proportion to the decrement of the total amount of integrin beta 1 subunits. When one transfectant (QA23) was cultured in a serum-free medium, cell shape changed from fibroblast-like to neuron-like morphology accompanied by a low growth rate, and the cells did not form focal contact on fibronectin. A similar morphological change occurred in QT6-c cells when the cells were infected with Rous Sarcoma virus, which could produce the Anti-Int. The QA23 cells did not attach to fibronectin as efficiently as did the original QT6-c cells. These data suggest that reduced expression of integrin beta 1 subunit affects cell growth as well as cell morphology by disordering the interaction between integrins and matrix proteins and/or cytoplasmic proteins.
Hayashi
Y
Y
Biochemical Research Institute, Morinaga Milk Ind. Co. Ltd., Kanagawa, Japan.
Iguchi
T
T
Kawashima
T
T
Bao
Z Z
ZZ
Yacky
C
C
Boettiger
D
D
Horwitz
A F
AF
D000818:Animals; D015152:Blotting, Northern; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D015870:Gene Expression / Q000235:genetics*; D005822:Genetic Vectors; D016023:Integrins / Q000235:genetics* / Q000378:metabolism; D046911:Macromolecular Substances; D011233:Precipitin Tests; D011784:Quail; D012313:RNA / Q000235:genetics / Q000378:metabolism; D016372:RNA, Antisense / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
D016895:Culture Media, Serum-Free; D005353:Fibronectins; D016023:Integrins; D046911:Macromolecular Substances; D016372:RNA, Antisense; D012313:RNA