Differential display and cloning of shear stress-responsive messenger RNAs in human endothelial cells.
To investigate the effect of shear stress on endothelial gene expression, we performed differential display of mRNAs from cultured human umbilical vein endothelial cells either incubated under static conditions or exposed to shear stress (15 dynes/cm2) for 6 h in a flow-chamber. Around 4% of the total number of mRNAs detected were either up- or down-regulated by shear stress. DNA sequencing of some of these shear stress-responsive mRNAs revealed homology of several clones to known gene sequences and many other clones for unknown genes. Known genes, including those for human laminin B1 chain, H(+)-ATP synthase coupling factor 6, lysyl oxidase, myosin light chain kinase, and interleukin-8 receptor, were upregulated by shear stress, while the gene encoding NADH dehydrogenase was down-regulated. The present results suggest that shear stress can change the expression of numerous genes in endothelial cells, far more than reported to date, and that mRNA differential display is quite useful for cloning known and unknown shear stress-responsive genes.
Ando
J
J
Department of Cardiovascular Biomechanics, Faculty of Medicine, University of Tokyo, Japan.
Tsuboi
H
H
Korenaga
R
R
Takahashi
K
K
Kosaki
K
K
Isshiki
M
M
Tojo
T
T
Takada
Y
Y
Kamiya
A
A
D015703:Antigens, CD / Q000235:genetics; D003001:Cloning, Molecular; D018076:DNA, Complementary; D015536:Down-Regulation; D004730:Endothelium, Vascular / Q000378:metabolism*; D006801:Humans; D007797:Laminin / Q000235:genetics; D009219:Myosin-Light-Chain Kinase / Q000235:genetics; D009245:NADH Dehydrogenase / Q000235:genetics; D008249:Protein-Lysine 6-Oxidase / Q000235:genetics; D006180:Proton-Translocating ATPases / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics*; D018123:Receptors, Interleukin / Q000235:genetics; D023062:Receptors, Interleukin-8A
D015703:Antigens, CD; D018076:DNA, Complementary; D007797:Laminin; D012333:RNA, Messenger; D018123:Receptors, Interleukin; D023062:Receptors, Interleukin-8A; D008249:Protein-Lysine 6-Oxidase; D009245:NADH Dehydrogenase; D009219:Myosin-Light-Chain Kinase; D006180:Proton-Translocating ATPases