PMID- 22773906
OWN - NLM
STAT- MEDLINE
DCOM- 20121113
LR  - 20211021
IS  - 1090-0535 (Electronic)
IS  - 1090-0535 (Linking)
VI  - 18
DP  - 2012
TI  - Upregulation of heparanase in high-glucose-treated endothelial cells promotes 
      endothelial cell migration and proliferation and correlates with Akt and 
      extracellular-signal-regulated kinase phosphorylation.
PG  - 1684-95
AB  - PURPOSE: The objectives of this study were to determine whether 
      high-glucose-induced upregulation of heparanase (HPSE) expression and 
      differential heparanase expression in human retinal vascular endothelial cells 
      (HRECs) can alter HREC migration and proliferation. We also aimed to determine 
      whether HREC migration and proliferation correlate with the levels of protein 
      kinase B (Akt) and extracellular-signal-regulated kinase (ERK) phosphorylation 
      and activation. METHODS: HRECs were treated with either 5 mM glucose (Glu5) or 
      high (30 mM) glucose (Glu30) for 48 h. Untransfected HRECs were grown in human 
      endothelial serum-free medium (HE-SFM) in the presence of 5 mM glucose and 
      supplemented with 30 mM mannitol for 48 h as an osmotic control (mannitol). HRECs 
      were also infected with a heparanase small interfering RNA recombinant lentiviral 
      vector (HPSE-LV) or a control vector (Con-LV) at a multiplicity of infection 
      (MOI) of 60 for three days. Then the con-LV and HPSE-LV-infected cells were 
      treated with 30 mM glucose for 48 h (Con-LV-Glu30 and HSPE-LV-Glu30, 
      respectively). The expression levels of heparanase mRNA and protein and HREC 
      proliferation and migration were examined using quantitative real-time polymerase 
      chain reaction (qRT-PCR), western blot analysis, 
      3-(4,5-dimethylthiahiazol-2-y1)-2,5-diphenyltetrazolium bromide assay, 
      bromodeoxyuridine histochemical staining, and the Boyden chamber assay. The 
      expression level of paxillin was examined using immunofluorescent staining. Akt 
      and ERK phosphorylation was evaluated using western blot analysis. RESULTS: We 
      successfully transfected the HPSE RNAi lentiviral vector into HRECs and 
      demonstrated that it can suppress the expression of the heparanase gene in these 
      cells. Western blot and qRT-PCR analyses showed that HRECs treated with a high 
      concentration of glucose exhibited increased heparanase protein and mRNA levels, 
      while the levels were decreased in HRECs that had been infected with HPSE-LV 
      before treatment with high glucose (HPSE-LV-Glu30; p<0.05). The observed increase 
      or decrease in the levels of heparanase correlated with increased or decreased 
      HREC migration and proliferation, respectively (p<0.05). HREC proliferation and 
      migration were found to correlate with Akt and ERK phosphorylation levels 
      (p<0.5). CONCLUSIONS: Our results indicate that heparanase plays a significant 
      role in mediating retinal vascular endothelial cell proliferation and migration 
      after the HRECs are exposed to high levels of glucose. Signaling inducing 
      heparanase-stimulated HREC proliferation and migration appears to be related to 
      the activation of Akt and ERK via their phosphorylation.
FAU - Yuan, Ling
AU  - Yuan L
AD  - State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic center of Sun Yat-sen 
      University, Guangzhou, China.
FAU - Hu, Jie
AU  - Hu J
FAU - Luo, Yan
AU  - Luo Y
FAU - Liu, QingYun
AU  - Liu Q
FAU - Li, Tao
AU  - Li T
FAU - Parish, Christopher R
AU  - Parish CR
FAU - Freeman, Craig
AU  - Freeman C
FAU - Zhu, XiaoBo
AU  - Zhu X
FAU - Ma, Wei
AU  - Ma W
FAU - Hu, XuTing
AU  - Hu X
FAU - Yu, HongHua
AU  - Yu H
FAU - Tang, ShiBo
AU  - Tang S
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20120620
PL  - United States
TA  - Mol Vis
JT  - Molecular vision
JID - 9605351
RN  - 0 (Culture Media, Serum-Free)
RN  - 0 (Paxillin)
RN  - 0 (RNA, Small Interfering)
RN  - 3OWL53L36A (Mannitol)
RN  - EC 2.7.11.1 (Proto-Oncogene Proteins c-akt)
RN  - EC 2.7.11.24 (Extracellular Signal-Regulated MAP Kinases)
RN  - EC 3.2.1.- (heparanase)
RN  - EC 3.2.1.31 (Glucuronidase)
RN  - IY9XDZ35W2 (Glucose)
SB  - IM
MH  - Cell Movement/drug effects
MH  - Cell Proliferation/drug effects
MH  - Culture Media, Serum-Free
MH  - Endothelial Cells/cytology/*drug effects/metabolism
MH  - Extracellular Signal-Regulated MAP Kinases/genetics/*metabolism
MH  - Genetic Vectors
MH  - Glucose/*pharmacology
MH  - Glucuronidase/antagonists & inhibitors/genetics/*metabolism
MH  - Humans
MH  - Lentivirus
MH  - Mannitol/chemistry
MH  - Paxillin/genetics/metabolism
MH  - Phosphorylation/drug effects
MH  - Primary Cell Culture
MH  - Proto-Oncogene Proteins c-akt/genetics/*metabolism
MH  - RNA, Small Interfering/genetics
MH  - Retina/cytology/*drug effects/metabolism
MH  - Signal Transduction/drug effects
MH  - Up-Regulation
MH  - Young Adult
PMC - PMC3388982
EDAT- 2012/07/10 06:00
MHDA- 2012/11/14 06:00
PMCR- 2012/01/01
CRDT- 2012/07/10 06:00
PHST- 2010/09/23 00:00 [received]
PHST- 2012/06/17 00:00 [accepted]
PHST- 2012/07/10 06:00 [entrez]
PHST- 2012/07/10 06:00 [pubmed]
PHST- 2012/11/14 06:00 [medline]
PHST- 2012/01/01 00:00 [pmc-release]
AID - 173 [pii]
PST - ppublish
SO  - Mol Vis. 2012;18:1684-95. Epub 2012 Jun 20.