Panax notoginseng saponins induced up-regulation, phosphorylation and binding activity of MEK, ERK, AKT, PI-3K protein kinases and GATA transcription factors in hematopoietic cells

Xin Sun; Rui-lan Gao; Xiao-jie Lin; Wei-hong Xu; Xiao-hong Chen

doi:10.1007/s11655-012-1306-4

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Panax notoginseng saponins induced up-regulation, phosphorylation and binding activity of MEK, ERK, AKT, PI-3K protein kinases and GATA transcription factors in hematopoietic cells

Updated：2021-08-27

- Panax notoginseng saponins induced up-regulation, phosphorylation and binding activity of MEK, ERK, AKT, PI-3K protein kinases and GATA transcription factors in hematopoietic cells
- Chinese Journal of Integrative Medicine Vol. 19, Issue 2, Pages: 112-118(2013)
- Affiliations：
  
  1. Department of Oncology, Zhejiang Provincial People’s Hospital,Hangzhou,China
  2. Institution of Hematology Research, The First Affiliated Hospital, Zhejiang Chinese Medical University,Hangzhou,China
- Author bio：
- Funds：
  
  Supported by National Natural Science Foundation of China (No. 30070933)
- DOI：10.1007/s11655-012-1306-4
  CLC：
- Published：2013，
  
  Published Online：31 January 2013，
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Sun, X., Gao, Rl., Lin, Xj. et al. Panax notoginseng saponins induced up-regulation, phosphorylation and binding activity of MEK, ERK, AKT, PI-3K protein kinases and GATA transcription factors in hematopoietic cells., Chin. J. Integr. Med. 19, 112–118 (2013). https://doi.org/10.1007/s11655-012-1306-4

Xin Sun, Rui-lan Gao, Xiao-jie Lin, et al. Panax notoginseng saponins induced up-regulation, phosphorylation and binding activity of MEK, ERK, AKT, PI-3K protein kinases and GATA transcription factors in hematopoietic cells. [J]. Chinese Journal of Integrative Medicine 19(2):112-118(2013)
Sun, X., Gao, Rl., Lin, Xj. et al. Panax notoginseng saponins induced up-regulation, phosphorylation and binding activity of MEK, ERK, AKT, PI-3K protein kinases and GATA transcription factors in hematopoietic cells., Chin. J. Integr. Med. 19, 112–118 (2013). https://doi.org/10.1007/s11655-012-1306-4 DOI：

Xin Sun, Rui-lan Gao, Xiao-jie Lin, et al. Panax notoginseng saponins induced up-regulation, phosphorylation and binding activity of MEK, ERK, AKT, PI-3K protein kinases and GATA transcription factors in hematopoietic cells. [J]. Chinese Journal of Integrative Medicine 19(2):112-118(2013) DOI： 10.1007/s11655-012-1306-4.

摘要

To investigate the effects of panax notoginseng saponins (PNS) on expression

regulation and phosphorylation of multiple protein kinases in mitogen activated protein kinase (MAPK) intracellular signal pathway and GATA transcription factors in hematopoietic cells

so as to explore its mechanism of proliferation and differentiation activity on hematopoiesis. The human granulocytic HL-60

erythrocytic K562

megakaryocytic CHRF-288 and Meg-01 cell lines were treated by PNS

the positive control of K562

CHRF-288 cells treated by recombination human erythropoietin (Epo) and thrombopoietin (Tpo) respectively. The total cell lysate and nuclei protein were extracted after being treated by PNS

subsequently

analyzed by both Western blot and immune-precipitation. Meanwhile

the nuclei extract was performed for electrophoretic mobility shift assay (EMSA) by using 32P radio labeled double-stranded GATA consensus oligonucleotide. The expression levels of kinase MEK-1

MEK-2

ERK-1

ERK-2

AKT-1

AKT-2 and PI-3K were increased by PNS treatment to different extent in four cell lines

depending on cellular heterogeneity and sensitivity to PNS

also phosphorylation of MEK-1

ERK-1 was differentially promoted by PNS respectively P<0.05

0.01

0.001). The expression levels of transcription factors GATA-1 and GATA-2 were increased

moreover

their DNA binding activities were raised dramatically in PNS treated K562

CHRF-288 and Meg-01 cells compared with the controls respectively (P<0.05

0.01

0.001). The positive control of K562

CHRF-288 cells treated by Epo or Tpo respectively also displayed up-regulation of protein kinases and GATA transcription factors respectively (P<0.05

0.01

0.001). The results indicated that intracellular signal pathway initiated by PNS was involved in MAPK pathway and transcription factors of GATA family in hematopoietic cells. PNS displayed the role to promote proliferation and differentiation

by means of increasing expression level and phosphorylation status of multiple protein kinases

also inducing synthesis of GATA transcription factors and upregulation its DNA binding activity.

Abstract

To investigate the effects of panax notoginseng saponins (PNS) on expression

regulation and phosphorylation of multiple protein kinases in mitogen activated protein kinase (MAPK) intracellular signal pathway and GATA transcription factors in hematopoietic cells

so as to explore its mechanism of proliferation and differentiation activity on hematopoiesis. The human granulocytic HL-60

erythrocytic K562

megakaryocytic CHRF-288 and Meg-01 cell lines were treated by PNS

the positive control of K562

CHRF-288 cells treated by recombination human erythropoietin (Epo) and thrombopoietin (Tpo) respectively. The total cell lysate and nuclei protein were extracted after being treated by PNS

subsequently

analyzed by both Western blot and immune-precipitation. Meanwhile

the nuclei extract was performed for electrophoretic mobility shift assay (EMSA) by using 32P radio labeled double-stranded GATA consensus oligonucleotide. The expression levels of kinase MEK-1

MEK-2

ERK-1

ERK-2

AKT-1

AKT-2 and PI-3K were increased by PNS treatment to different extent in four cell lines

depending on cellular heterogeneity and sensitivity to PNS

also phosphorylation of MEK-1

ERK-1 was differentially promoted by PNS respectively P<0.05

0.01

0.001). The expression levels of transcription factors GATA-1 and GATA-2 were increased

moreover

their DNA binding activities were raised dramatically in PNS treated K562

CHRF-288 and Meg-01 cells compared with the controls respectively (P<0.05

0.01

0.001). The positive control of K562

CHRF-288 cells treated by Epo or Tpo respectively also displayed up-regulation of protein kinases and GATA transcription factors respectively (P<0.05

0.01

by means of increasing expression level and phosphorylation status of multiple protein kinases

also inducing synthesis of GATA transcription factors and upregulation its DNA binding activity.

关键词

panax notoginseng saponinshematopoietic cellsprotein kinasephosphorylationtranscription factor

Keywords

panax notoginseng saponinshematopoietic cellsprotein kinasephosphorylationtranscription factor

references

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Qian XD, Gao RL, Ke M, Jin JM. Proliferation and differentiation of human CD34+ hematopoietic stem/progenitor cells induced by panax notoginosides. Chin J Exp Hematol (Chin) 2003;11:120–123.

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Gai Y, Gao RL, Niu YP. Effect of panax notoginsenosides on the proliferation of hematopoietic progenitor cells in mice with immune-mediated aplastic anemia. Chin J Integr Tradit West Med (Chin) 2003;23:680–683.

Gao RL, Lin XJ, Chen XH. Experimental study of hematopoietic cell gene expression profile induced by Panax Notoginsenosides in vitro. Chin J Integr Tradit West Med (Chin) 2007;27:999–1003.

Zhang Y, Dong C. Regulatory mechanisms of mitogen-activated kinase signaling. Cell Mol Life Sci 2007;64:2771–2789.

Millikan PD, Balamohan SM, Raskind WH, Kacena MA. Inherited thrombocytopenia due to GATA-1 mutations. Semin Thromb Hemost 2011;37:682–689.

Bresnick EH, Lee HY, Fujiwara T, Johnson KD, Keles S. GATA switches as developmental drivers. J Biol Chem 2010;28:31087–31093.

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Institute of Hematology, the First Hospital Affiliated to Zhejiang University of Chinese Medicine

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