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Activation of Notch3 promotes pulmonary arterial smooth muscle cells proliferation via Hes1/p27Kip1 signaling pathway.

Song Y, Zhang Y, Jiang H, Zhu Y, Liu L, Feng W, Yang L, Wang Y, Li M - FEBS Open Bio (2015)

Bottom Line: However, the detailed molecular mechanisms underlying this effect are still unclear.The present study aims to address this issue.This was accompanied with up-regulation of Hes1 protein and down-regulation of p27Kip1 protein.

View Article: PubMed Central - PubMed

Affiliation: Department of Respiratory Medicine, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, PR China.

ABSTRACT
Activation of the Notch3 cascade is involved in the development of pulmonary arterial hypertension by stimulating the proliferation of vascular smooth muscle cells. However, the detailed molecular mechanisms underlying this effect are still unclear. The present study aims to address this issue. We demonstrated that over-expression of intracellular domain of the Notch3 receptor (NICD3) by adenovirus transfection dramatically induced proliferation of primary cultured pulmonary artery smooth muscle cells. This was accompanied with up-regulation of Hes1 protein and down-regulation of p27Kip1 protein. More importantly, we observed that prior silencing of Hes1 with siRNA blocked NICD3 over-expression-induced p27Kip1 reduction and cell proliferation. The present study suggests that Hes1 lies downstream of NICD3 and particularly mediates Notch3 signaling-induced proliferation of pulmonary arterial smooth muscle cells by down-regulation of p27Kip1 expression.

No MeSH data available.


Related in: MedlinePlus

Over-expression of NICD3 induces PASMCs proliferation. (A) The representative Western blot of NICD3 over-expression by adenovirus transfection in primary cultured PASMCs (n = 3). (B) Over-expression of NICD3 induced PASMCs proliferation assessed by BrdU incorporation assay (n = 4 each group). *P < 0.01 versus control cells.
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f0005: Over-expression of NICD3 induces PASMCs proliferation. (A) The representative Western blot of NICD3 over-expression by adenovirus transfection in primary cultured PASMCs (n = 3). (B) Over-expression of NICD3 induced PASMCs proliferation assessed by BrdU incorporation assay (n = 4 each group). *P < 0.01 versus control cells.

Mentions: To determine whether activation of Notch3 induces PASMCs proliferation, NICD3 was over-expressed in cells by adenovirus transfection. Fig. 1A shows that control PASMCs had almost no basal NICD3 protein, and adenovirus transfection did not increase NICD3 protein level. PASMCs transfected with adenovirus carrying NICD3 showed a dramatic increase in NICD3 protein level. Fig. 1B indicates that cells with over-expressed NICD3 exhibited an elevated proliferation assessed by BrdU incorporation assay, which was 3-fold increase over control (P < 0.01), while transfection of cells with adenovirus did not affect cells proliferation. These results suggest that activation of Notch3 effectively induces PASMCs proliferation.


Activation of Notch3 promotes pulmonary arterial smooth muscle cells proliferation via Hes1/p27Kip1 signaling pathway.

Song Y, Zhang Y, Jiang H, Zhu Y, Liu L, Feng W, Yang L, Wang Y, Li M - FEBS Open Bio (2015)

Over-expression of NICD3 induces PASMCs proliferation. (A) The representative Western blot of NICD3 over-expression by adenovirus transfection in primary cultured PASMCs (n = 3). (B) Over-expression of NICD3 induced PASMCs proliferation assessed by BrdU incorporation assay (n = 4 each group). *P < 0.01 versus control cells.
© Copyright Policy - CC BY
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4556730&req=5

f0005: Over-expression of NICD3 induces PASMCs proliferation. (A) The representative Western blot of NICD3 over-expression by adenovirus transfection in primary cultured PASMCs (n = 3). (B) Over-expression of NICD3 induced PASMCs proliferation assessed by BrdU incorporation assay (n = 4 each group). *P < 0.01 versus control cells.
Mentions: To determine whether activation of Notch3 induces PASMCs proliferation, NICD3 was over-expressed in cells by adenovirus transfection. Fig. 1A shows that control PASMCs had almost no basal NICD3 protein, and adenovirus transfection did not increase NICD3 protein level. PASMCs transfected with adenovirus carrying NICD3 showed a dramatic increase in NICD3 protein level. Fig. 1B indicates that cells with over-expressed NICD3 exhibited an elevated proliferation assessed by BrdU incorporation assay, which was 3-fold increase over control (P < 0.01), while transfection of cells with adenovirus did not affect cells proliferation. These results suggest that activation of Notch3 effectively induces PASMCs proliferation.

Bottom Line: However, the detailed molecular mechanisms underlying this effect are still unclear.The present study aims to address this issue.This was accompanied with up-regulation of Hes1 protein and down-regulation of p27Kip1 protein.

View Article: PubMed Central - PubMed

Affiliation: Department of Respiratory Medicine, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, PR China.

ABSTRACT
Activation of the Notch3 cascade is involved in the development of pulmonary arterial hypertension by stimulating the proliferation of vascular smooth muscle cells. However, the detailed molecular mechanisms underlying this effect are still unclear. The present study aims to address this issue. We demonstrated that over-expression of intracellular domain of the Notch3 receptor (NICD3) by adenovirus transfection dramatically induced proliferation of primary cultured pulmonary artery smooth muscle cells. This was accompanied with up-regulation of Hes1 protein and down-regulation of p27Kip1 protein. More importantly, we observed that prior silencing of Hes1 with siRNA blocked NICD3 over-expression-induced p27Kip1 reduction and cell proliferation. The present study suggests that Hes1 lies downstream of NICD3 and particularly mediates Notch3 signaling-induced proliferation of pulmonary arterial smooth muscle cells by down-regulation of p27Kip1 expression.

No MeSH data available.


Related in: MedlinePlus