CREG inhibits migration of human vascular smooth muscle cells by mediating IGF-II endocytosis.
We previously determined that the cellular repressor of E1A-stimulated genes, (CREG) plays a role in the maintenance of the mature phenotype of vascular smooth muscle cells (SMCs). This study aimed to identify the role of CREG in modulating the migration of SMCs. Recombinant virus-mediated CREG expression inhibited the cellular migration of cultured SMCs associated with down-regulated activity of matrix metalloproteinase-9 (MMP-9). In contrast, CREG knockdown via the retroviral transfer of short hairpin RNAs promoted cellular migration. Enzyme-linked immunosorbent assay and endocytosis analysis revealed that CREG knockdown attenuated the internalization and increased secretion of insulin-like growth factor (IGF)-II. Western blot analysis demonstrated that both phosphoinositide 3-kinase (PI3K) and phosphatase Akt were enhanced in CREG knockdown SMCs. Furthermore, the effect of CREG knockdown on SMC migration was abrogated in a dose-dependent manner by the addition of either IGF-II neutralizing antibody or the PI3K inhibitor, LY294002. These results indicate that the CREG knockdown-mediated increase in IGF-II secretion promoted cellular migration in SMCs via the PI3K/Akt signal pathway. Additionally, blockage of IGF-II binding to the mannose-6-phosphate/IGF-II receptor (M6P/IGF2R) by IGF2R antibody or recombinant IGF2R fragment attenuated the endocytosis of IGF-II in cells overexpressing CREG. This indicates that M6P/IGF2R is involved in the regulation of CREG-mediated IGF-II endocytosis. In summary, these data demonstrate for the first time that CREG plays a critical role in the inhibition of SMC migration, as well as maintaining SMCs in a mature phenotype. These results may provide a new therapeutic target for vascular disease associated with neointimal hyperplasia.
Han
Yaling
Y
Cardiovascular Research Institute, Department of Cardiology, Shenyang Northern Hospital, Shenyang, China. hanyaling@263.net
Cui
Jifu
J
Tao
Jie
J
Guo
Liang
L
Guo
Peng
P
Sun
Mingyu
M
Kang
Jian
J
Zhang
Xiaolin
X
Yan
Chenghui
C
Li
Shaohua
S
D002465:Cell Movement; D002478:Cells, Cultured; D004705:Endocytosis; D006801:Humans; D007335:Insulin-Like Growth Factor II / Q000378:metabolism*; D008360:Mannosephosphates / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D032389:Myocytes, Smooth Muscle / Q000166:cytology*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D034741:RNA, Small Interfering / Q000494:pharmacology; D017527:Receptor, IGF Type 2 / Q000378:metabolism; D012097:Repressor Proteins / Q000502:physiology*; D015398:Signal Transduction
C114068:CREG1 protein, human; D008360:Mannosephosphates; D034741:RNA, Small Interfering; D017527:Receptor, IGF Type 2; D012097:Repressor Proteins; C027693:mannose-6-phosphate; D007335:Insulin-Like Growth Factor II; D019869:Phosphatidylinositol 3-Kinases