Zhen-Zhong Zheng1, Zheng-Xiang Liu. 1. Department of Cardiology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Abstract
AIM: To investigate the effects of CD151 on the activity of endothelial NO synthase (eNOS), and ECV304 migration, proliferation and tube formation. METHODS: pAAV-CD151 and pAAV-anti-CD151 were constructed and used to transiently transfect ECV304 mediated with Lipofectamine 2000. After transfection, the expression of CD151 was measured by Western blotting. Cell migration assay was performed using Boyden transwell; proliferation assay was evaluated using the 3- [4,5-dimethylthiazol-2-yl]-2,5, diphenyltetrazolium bromide (MTT) method, and tube formation test was examined on matrigel. eNOS activity was assayed by L- [3H]citrulline production from L-[3H]arginine. The involvement of eNOS was explored using an eNOS inhibitor (L-NAME) and the effects in the process were observed. RESULTS: CD151 promotes cell migration, proliferation and tube formation. In addition, CD151 increases eNOS activity. Moreover, cell migration, proliferation and tube formation induced by CD151 are inhibited when L-NAME is used, which indicates that there is an involvement of eNOS in CD151-induced cell migration, cell proliferation and tube formation. CONCLUSION: CD151 promotes ECV304 migration, proliferation and tube formation. The mechanism is that CD151 increases eNOS activity. This result also suggests that eNOS is involved in the angiogenic effects of CD151.
AIM: To investigate the effects of CD151 on the activity of endothelial NO synthase (eNOS), and ECV304 migration, proliferation and tube formation. METHODS:pAAV-CD151 and pAAV-anti-CD151 were constructed and used to transiently transfect ECV304 mediated with Lipofectamine 2000. After transfection, the expression of CD151 was measured by Western blotting. Cell migration assay was performed using Boyden transwell; proliferation assay was evaluated using the 3- [4,5-dimethylthiazol-2-yl]-2,5, diphenyltetrazolium bromide (MTT) method, and tube formation test was examined on matrigel. eNOS activity was assayed by L- [3H]citrulline production from L-[3H]arginine. The involvement of eNOS was explored using an eNOS inhibitor (L-NAME) and the effects in the process were observed. RESULTS:CD151 promotes cell migration, proliferation and tube formation. In addition, CD151 increases eNOS activity. Moreover, cell migration, proliferation and tube formation induced by CD151 are inhibited when L-NAME is used, which indicates that there is an involvement of eNOS in CD151-induced cell migration, cell proliferation and tube formation. CONCLUSION:CD151 promotes ECV304 migration, proliferation and tube formation. The mechanism is that CD151 increases eNOS activity. This result also suggests that eNOS is involved in the angiogenic effects of CD151.