Acta Pharmacologica Sinica (2010) 31: 805–812; doi: 10.1038/aps.2010.65

Acta Pharmacologica Sinica (2010) 31: 805–812; doi: 10.1038/aps.2010.65; published online 28 June 2010

Original Article

Activation of the ERK signaling pathway is involved in CD151-induced angiogenic effects on the formation of CD151-integrin complexes

Hou-juan ZUO^#, Jing-yang LIN^{#, $}, Zhao-yu LIU, Wei-feng LIU, Tao LIU, Jun YANG, Yu LIU, Dao-wen WANG, Zheng-xiang LIU*

Department of Cardiology of Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China

Aim: To assess the roles of extracellular signal-regulated kinase (ERK), p38, and CD151-integrin complexes on proliferation, migration, and tube formation activities of CD151-induced human umbilical vein endothelial cells (HUVECs).

Methods: CD151, anti-CD151 and CD151-AAA mutant were inserted into recombinant adeno-associated virus (rAAV) vectors and used to transfect HUVECs. After transfection, the expression of CD151 was measured. Proliferation was assessed using the 3-[4,5-dimethylthiazol- 2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay. Cell migration was evaluated in Boyden transwell chambers using FBS as the chemotactic stimulus. The tube formation assay was performed on matrigel. The potential involvement of various signaling pathways was explored using selective inhibitors.

Results: CD151 gene delivery increased the expression of CD151 at both the mRNA and protein levels. Overexpression of CD151 promoted cell proliferation, migration and tube formation in vitro, and phosphorylation of ERK was also increased. Further, CD151-induced cell proliferation, migration, and tube formation were attenuated by the ERK inhibitor PD98059 (20 µmol/L) but not by a p38 inhibitor (SB203580, 20 µmol/L). Moreover, there was no significant difference in CD151 protein expression between the CD151 group and the CD151-AAA group, but the CD151-AAA mutant abrogated cellular proliferation, migration, and tube formation and decreased the phosphorylation of ERK.

Conclusion: This study suggests that activation of the ERK signaling pathway may be involved in the angiogenic effects of CD151. Activation of ERK was dependent on the formation of CD151-integrin complexes. Therefore modulation of CD151 may be as a novel therapeutic strategy for regulating angiogenesis.

Keywords: CD151; cell migration; cell proliferation; angiogenesis; ERK; p38 MAPK; human umbilical vein endothelial cells

The project was supported by a grant from the National Natural Science Foundation of China (No 30670856).

We are grateful to Dr Xin ZHANG for providing the PzeoSV-CD151 plasmid (Department of Molecular Science, University of Tennessee Health Science Center).

^# These two authors contributed equally to this work.
^$ Now in Department of Cardiology of Zhejiang Provincial People’s Hospital, Hangzhou China
* To whom correspondence should be addressed.
E-mail liuzhengxiang@hotmail.com
Received 2009-10-05 Accepted 2010-05-07

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