Dysregulation of CREB binding protein triggers thrombin-induced proliferation of vascular smooth muscle cells.
Thrombin is a potent mitogen for vascular smooth muscle cells (VSMCs). CBP has been regarded as a potential therapeutic target on the basis of its ability to affect cell growth. Therefore we hypothesized that CBP mediates thrombin-induced proliferation of VSMCs. We constructed recombinant adenoviral vector that expresses four short hairpin RNA (shRNA) targeting rat CBP mRNA (CBP-shRNA/Ad). VSMCs were infected with CBP-shRNA/Ad and treated with thrombin. CBP level were analyzed by quantitative real-time PCR and Western blot. To evaluate VSMC proliferation, the cell cycle and DNA synthesis were analyzed by flow cytometry and (3)H-thymidine incorporation, respectively. CBP-shRNA/Ad infection inhibited thrombin-induced CBP expression in a dose-dependent manner concomitant with a decrease in the percentage of cells in the S phase and in DNA synthesis. These findings suggest that CBP plays a pivotal role in the S phase progression of VSMCs.
Chen
Jing
J
Department of Cardiology, Renmin Hospital, Wuhan University, Wuhan, China.
Jiang
Hong
H
Xu
Lin
L
Zhu
Li-Hua
LH
Wang
Lang
L
Wen
Hua-Zhi
HZ
Hu
Xiao-Rong
XR
D000256:Adenoviridae; D000818:Animals; D050882:CREB-Binding Protein / Q000378:metabolism*; D002453:Cell Cycle / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D004247:DNA / Q000096:biosynthesis; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D032389:Myocytes, Smooth Muscle / Q000166:cytology* / Q000187:drug effects*; D034741:RNA, Small Interfering / Q000378:metabolism; D051381:Rats; D013917:Thrombin / Q000494:pharmacology*
D034741:RNA, Small Interfering; D004247:DNA; D050882:CREB-Binding Protein; C494365:Crebbp protein, rat; D013917:Thrombin