PMID- 21851826
OWN - NLM
STAT- MEDLINE
DCOM- 20111122
LR  - 20151119
IS  - 1879-0631 (Electronic)
IS  - 0024-3205 (Linking)
VI  - 89
IP  - 17-18
DP  - 2011 Oct 24
TI  - Sildenafil inhibits calcineurin/NFATc2-mediated cyclin A expression in pulmonary 
      artery smooth muscle cells.
PG  - 644-9
LID - 10.1016/j.lfs.2011.07.023 [doi]
AB  - AIMS: To examine whether calcineurin/NFAT signaling pathway leads to 
      proliferation of pulmonary artery smooth muscle cells (PASMCs) by regulating cell 
      cycle proteins and whether the phosphodiesterase-5 (PDE5) inhibitor sildenafil 
      affects calcineurin/NFAT-induced cell proliferation. MAIN METHODS: A 
      [(3)H]thymidine incorporation assay was used to examine DNA synthesis (cell 
      proliferation); cyclin A and NFATc2 expressions were determined by Western blot. 
      Cyclin-dependent kinase 2 (CDK2) activity was measured with an in vitro kinase 
      activity assay, and calcineurin and NFAT activity were evaluated using a 
      calcineurin assay kit and a luciferase activity assay, respectively. A chemical 
      inhibitor or siRNA transfection was used to inhibit calcineurin/NFAT signaling 
      pathway. KEY FINDINGS: Serotonin dose-dependently stimulated cyclin A expression 
      in PASMCs. This effect was accompanied by dose-dependent increases in CDK2 
      activity and the rate of DNA synthesis. At the same time, PASMCs treated with 
      serotonin showed dose-dependent activation of calcineurin/NFAT signaling pathway. 
      Inhibition of calcineurin activity by cyclosporine A or loss of NFATc2 protein by 
      siRNA transfection abolished serotonin-induced cyclin A expression and consequent 
      CDK2 activation and DNA synthesis. We further found that pretreatment of cells 
      with sildenafil suppressed serotonin-triggered activation of calcineurin/NFATc2 
      signaling pathway and resultant cyclin A expression, CDK2 activation and cell 
      proliferation, while the presence of DT-3 [a specific protein kinase G (PKG) 
      peptide inhibitor] reversed the effects of sildenafil on PASMCs. SIGNIFICANCE: 
      Our study suggests that enhanced PKG activity suppresses calcineurin/NFATc2 
      cascade-mediated cyclin A expression, CDK2 activation and PASMC proliferation to 
      contribute to the overall effects of sildenafil in the treatment of pulmonary 
      hypertension.
CI  - Copyright (c) 2011 Elsevier Inc. All rights reserved.
FAU - Li, Manxiang
AU  - Li M
AD  - Respiratory Diseases Research Center, The Second Affiliated Hospital of Medical 
      College, Xi'an Jiaotong University, Xi'an, Shaanxi, 710004, PR China. 
      manxiangli@hootmail.com
FAU - Liu, Yuan
AU  - Liu Y
FAU - Sun, Xiuzhen
AU  - Sun X
FAU - Li, Zongfang
AU  - Li Z
FAU - Liu, Yun
AU  - Liu Y
FAU - Fang, Ping
AU  - Fang P
FAU - He, Ping
AU  - He P
FAU - Shi, Hongyang
AU  - Shi H
FAU - Xie, Mei
AU  - Xie M
FAU - Wang, Xiaochuang
AU  - Wang X
FAU - Zhang, Dexin
AU  - Zhang D
FAU - Zhang, Yonghong
AU  - Zhang Y
FAU - Ming, Zongjuan
AU  - Ming Z
FAU - Xu, Jing
AU  - Xu J
FAU - Lu, Jiamei
AU  - Lu J
FAU - Xie, Xinming
AU  - Xie X
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20110809
PL  - Netherlands
TA  - Life Sci
JT  - Life sciences
JID - 0375521
RN  - 0 (Cyclin A)
RN  - 0 (NFATC Transcription Factors)
RN  - 0 (Phosphodiesterase 5 Inhibitors)
RN  - 0 (Piperazines)
RN  - 0 (Purines)
RN  - 0 (RNA, Small Interfering)
RN  - 0 (Sulfones)
RN  - 333DO1RDJY (Serotonin)
RN  - BW9B0ZE037 (Sildenafil Citrate)
RN  - EC 2.7.11.22 (Cyclin-Dependent Kinase 2)
RN  - EC 3.1.3.16 (Calcineurin)
SB  - IM
MH  - Animals
MH  - Calcineurin/genetics/*metabolism
MH  - Cell Proliferation/drug effects
MH  - Cells, Cultured
MH  - Cyclin A/*genetics
MH  - Cyclin-Dependent Kinase 2/genetics/metabolism
MH  - Gene Expression Regulation/drug effects
MH  - Myocytes, Smooth Muscle/*drug effects/metabolism
MH  - NFATC Transcription Factors/genetics/*metabolism
MH  - Phosphodiesterase 5 Inhibitors/*pharmacology
MH  - Piperazines/*pharmacology
MH  - Pulmonary Artery/*cytology/drug effects
MH  - Purines/pharmacology
MH  - RNA, Small Interfering/genetics
MH  - Rats
MH  - Rats, Sprague-Dawley
MH  - Serotonin/metabolism
MH  - Signal Transduction/drug effects
MH  - Sildenafil Citrate
MH  - Sulfones/*pharmacology
EDAT- 2011/08/20 06:00
MHDA- 2011/12/13 00:00
CRDT- 2011/08/20 06:00
PHST- 2010/11/15 00:00 [received]
PHST- 2011/07/21 00:00 [revised]
PHST- 2011/07/28 00:00 [accepted]
PHST- 2011/08/20 06:00 [entrez]
PHST- 2011/08/20 06:00 [pubmed]
PHST- 2011/12/13 00:00 [medline]
AID - S0024-3205(11)00369-9 [pii]
AID - 10.1016/j.lfs.2011.07.023 [doi]
PST - ppublish
SO  - Life Sci. 2011 Oct 24;89(17-18):644-9. doi: 10.1016/j.lfs.2011.07.023. Epub 2011 
      Aug 9.