CD151 gene delivery increases eNOS activity and induces ECV304 migration, proliferation and tube formation
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.
Keywords:
(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.