Large scale isolation of non-uniform shear stress-responsive genes from cultured human endothelial cells through the preparation of a subtracted cDNA library

Hajime Yoshisue, Keiko Suzuki, Ayako Kawabata, Takeshi Ohya, Hanjun Zhao, Kazuhiro Sakurada, Yoji Taba, Toshiyuki Sasaguri, Naohiko Sakai, Shizuya Yamashita, Yuji Matsuzawa, Hiroshi Nojima

Research output: Contribution to journalArticle

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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.

Original languageEnglish
Pages (from-to)323-334
Number of pages12
JournalAtherosclerosis
Volume162
Issue number2
DOIs
Publication statusPublished - May 11 2002

Fingerprint

Gene Library
Endothelial Cells
Genes
Atherosclerosis
Human Umbilical Vein Endothelial Cells
Libraries
Plasminogen Activator Inhibitor 1
Interleukin-8
Aorta
Down-Regulation

All Science Journal Classification (ASJC) codes

  • Cardiology and Cardiovascular Medicine

Cite this

Large scale isolation of non-uniform shear stress-responsive genes from cultured human endothelial cells through the preparation of a subtracted cDNA library. / Yoshisue, Hajime; Suzuki, Keiko; Kawabata, Ayako; Ohya, Takeshi; Zhao, Hanjun; Sakurada, Kazuhiro; Taba, Yoji; Sasaguri, Toshiyuki; Sakai, Naohiko; Yamashita, Shizuya; Matsuzawa, Yuji; Nojima, Hiroshi.

In: Atherosclerosis, Vol. 162, No. 2, 11.05.2002, p. 323-334.

Research output: Contribution to journalArticle

Yoshisue, H, Suzuki, K, Kawabata, A, Ohya, T, Zhao, H, Sakurada, K, Taba, Y, Sasaguri, T, Sakai, N, Yamashita, S, Matsuzawa, Y & Nojima, H 2002, 'Large scale isolation of non-uniform shear stress-responsive genes from cultured human endothelial cells through the preparation of a subtracted cDNA library', Atherosclerosis, vol. 162, no. 2, pp. 323-334. https://doi.org/10.1016/S0021-9150(01)00735-3
Yoshisue, Hajime ; Suzuki, Keiko ; Kawabata, Ayako ; Ohya, Takeshi ; Zhao, Hanjun ; Sakurada, Kazuhiro ; Taba, Yoji ; Sasaguri, Toshiyuki ; Sakai, Naohiko ; Yamashita, Shizuya ; Matsuzawa, Yuji ; Nojima, Hiroshi. / Large scale isolation of non-uniform shear stress-responsive genes from cultured human endothelial cells through the preparation of a subtracted cDNA library. In: Atherosclerosis. 2002 ; Vol. 162, No. 2. pp. 323-334.
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