title: Large scale isolation of non-uniform shear stress-responsive genes from cultured human endothelial cells through the preparation of a subtracted cDNA library.
abstract: To investigate the molecular mechanisms responsible for the regional selectivity of early atherogenesis, we have applied a non-uniform shear stress to cultured human umbilical vein endothelial cells (HUVEC). We used a microcarrier culture system and a combination of subtraction and reverse-subtraction methods to isolate a number of genes upregulated by shear stress. The resultant subtracted library includes several known genes (e.g. MCP-1, TM) whose responsiveness to shear stress has been previously reported, indicating that the library is enriched for genes upregulated by shear stress. Also included are atherosclerosis-related genes (e.g. CTGF, IL-8) whose responsiveness to shear stress had not been demonstrated, other known genes whose relationship to atherosclerosis had not been reported, and novel genes. Some responsive to centrifugal force and shear stress (RECS) genes are also upregulated following stimulation by steady laminar shear stress in a parallel plate chamber. Interestingly, the library includes ET-1 and PAI, which are well known atherogenic factors that are downregulated by laminar shear stress. This implies that turbulent shear stress has effects on HUVEC that are different from those elicited by laminar shear stress. Importantly, analysis of specimens taken from human aorta showed that several RECS genes are transcriptionally upregulated in atherosclerotic lesions, suggesting that the subtracted library includes novel therapeutic targets for the treatment of atherosclerosis.
authors: Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., 3-6-6 Asahi-machi, Machida, 194-8533, Tokyo, Japan.
authors: Hajime
authors: H
authors: Yoshisue
authors: Keiko
authors: K
authors: Suzuki
authors: Ayako
authors: A
authors: Kawabata
authors: Takeshi
authors: T
authors: Ohya
authors: Hanjun
authors: H
authors: Zhao
authors: Kazuhiro
authors: K
authors: Sakurada
authors: Yoji
authors: Y
authors: Taba
authors: Toshiyuki
authors: T
authors: Sasaguri
authors: Naohiko
authors: N
authors: Sakai
authors: Shizuya
authors: S
authors: Yamashita
authors: Yuji
authors: Y
authors: Matsuzawa
authors: Hiroshi
authors: H
authors: Nojima
mesh_terms: D001011:Aorta / Q000166:cytology / Q000502:physiology; D001018:Aortic Diseases / Q000235:genetics; D001161:Arteriosclerosis / Q000235:genetics; D002478:Cells, Cultured; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology*; D015723:Gene Library; D006801:Humans; D013314:Stress, Mechanical; D015854:Up-Regulation
chemical_list: D018076:DNA, Complementary