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Tyrosine 23 Phosphorylation of Annexin A2 Promotes Proliferation, Invasion, and Stat3 Phosphorylation in the Nucleus of Human Breast Cancer SK-BR-3 Cells.

Wang YQ, Zhang F, Tian R, Ji W, Zhou Y, Sun XM, Liu Y, Wang ZY, Niu RF - Cancer Biol Med (2012)

Bottom Line: The monoclonal strains were screened.The monoclonal strains constitutively expressing Anxa2-WT, Anxa2-Y23A, and Anxa2-Y23D were screened.Immunoprecipitation analysis revealed that Anxa2 and Stat3 interacted with each other, and the expression of Stat3 phosphorylation in the nucleus was enhanced by Anxa2-Y23D.

View Article: PubMed Central - PubMed

Affiliation: Tianjin Medical University Cancer Institute and Hospital; Key Laboratory of Cancer Prevention and Therapy, Tianjin; Key Laboratory of Breast Cancer Prevention and Therapy, Ministry of Education, Tianjin 300060, China.

ABSTRACT

Objective: To investigate the role of tyrosine 23 (Tyr23) phosphorylation of Annexin A2 (Anxa2) in regulating the proliferation and invasion of human breast cancer SK-BR-3 cells.

Methods: A panel of lentivirus plasmids expressing Anxa2-wide type (Ana2-WT), Anxa2-Y23A, and Anxa2-Y23D was generated and infected with SK-BR-3 cells. The monoclonal strains were screened. The expression of Anxa2-WT, Anxa2-Y23A, and Anxa2-Y23D was determined by Western blot analysis. The ability of the cells to proliferate was detected through an MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] test. Boyden chamber assays were employed to examine migration and invasion abilities. The interaction between Anxa2 and Stat3 was analyzed by immunoprecipitation analyses. Nucleoprotein and cytosolic protein were extracted from SK-BR-3, Anxa2-WT, Anxa2-Y23A, and Anxa2-Y23D cells to analyze the expression and localization of Stat3 phosphorylation.

Results: The monoclonal strains constitutively expressing Anxa2-WT, Anxa2-Y23A, and Anxa2-Y23D were screened. Both Anxa2-WT and Anxa2-Y23D enhanced the proliferation, migration and invasion abilities of SK-BR-3 cells (P<0.05). Immunoprecipitation analysis revealed that Anxa2 and Stat3 interacted with each other, and the expression of Stat3 phosphorylation in the nucleus was enhanced by Anxa2-Y23D.

Conclusions: Tyr23 phosphorylation of Anxa2 promotes the proliferation and invasion of human breast cancer SK-BR-3 cells and the phosphorylation of Stat3 in the nucleus.

No MeSH data available.


Related in: MedlinePlus

Anxa2-WT and Anxa2-Y23D enhanced the proliferation of SK-BR-3 cells. A: Western blot analysis of GFP in SK-BR-3, control, Anxa2-WT, Anxa2-Y23A and Anxa2-Y23D cells. B: Immunofluorescence study of Anxa2-WT. Anxa2 was evidently overexpressed in SK-BR-3-Anxa2 cells. C: Proliferation ability was markedly enhanced in Anxa2-WT and Anxa2-Y23D cells. Triplicate experiments were done for B and C. The statistical significance was assessed by one-way ANOVA.
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f2: Anxa2-WT and Anxa2-Y23D enhanced the proliferation of SK-BR-3 cells. A: Western blot analysis of GFP in SK-BR-3, control, Anxa2-WT, Anxa2-Y23A and Anxa2-Y23D cells. B: Immunofluorescence study of Anxa2-WT. Anxa2 was evidently overexpressed in SK-BR-3-Anxa2 cells. C: Proliferation ability was markedly enhanced in Anxa2-WT and Anxa2-Y23D cells. Triplicate experiments were done for B and C. The statistical significance was assessed by one-way ANOVA.

Mentions: Lentivirus plasmids expressing Anxa2-WT, Anxa2-Y23A, or Anxa2-Y23D were infected with SK-BR-3 cells, and the strains that constitutively expressing these proteins were screened. Western blots were stained for green fluorescent protein (GFP) to determine the exact expression of Anxa2-WT, Anxa2-Y23A, and Anxa2-Y23D proteins in the SK-BR-3 cells (Figure 2A). Immunofluorescence studies of Anxa2 also showed an overexpression of Anxa2 in the SK-BR-3 cells, as depicted in Figure 2B. MTT assay results showed that compared with SK-BR-3, control (SK-BR-3 cells expressing empty pCDH lentivirus), and Anxa2-Y23A cells, the Anxa2-WT and Anxa2-Y23D cells exhibited an enhanced ability of duplication (Figure 2C). Anxa2-WT and Anxa2-Y23D enhanced the proliferation of SK-BR-3 cells.


Tyrosine 23 Phosphorylation of Annexin A2 Promotes Proliferation, Invasion, and Stat3 Phosphorylation in the Nucleus of Human Breast Cancer SK-BR-3 Cells.

Wang YQ, Zhang F, Tian R, Ji W, Zhou Y, Sun XM, Liu Y, Wang ZY, Niu RF - Cancer Biol Med (2012)

Anxa2-WT and Anxa2-Y23D enhanced the proliferation of SK-BR-3 cells. A: Western blot analysis of GFP in SK-BR-3, control, Anxa2-WT, Anxa2-Y23A and Anxa2-Y23D cells. B: Immunofluorescence study of Anxa2-WT. Anxa2 was evidently overexpressed in SK-BR-3-Anxa2 cells. C: Proliferation ability was markedly enhanced in Anxa2-WT and Anxa2-Y23D cells. Triplicate experiments were done for B and C. The statistical significance was assessed by one-way ANOVA.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3643677&req=5

f2: Anxa2-WT and Anxa2-Y23D enhanced the proliferation of SK-BR-3 cells. A: Western blot analysis of GFP in SK-BR-3, control, Anxa2-WT, Anxa2-Y23A and Anxa2-Y23D cells. B: Immunofluorescence study of Anxa2-WT. Anxa2 was evidently overexpressed in SK-BR-3-Anxa2 cells. C: Proliferation ability was markedly enhanced in Anxa2-WT and Anxa2-Y23D cells. Triplicate experiments were done for B and C. The statistical significance was assessed by one-way ANOVA.
Mentions: Lentivirus plasmids expressing Anxa2-WT, Anxa2-Y23A, or Anxa2-Y23D were infected with SK-BR-3 cells, and the strains that constitutively expressing these proteins were screened. Western blots were stained for green fluorescent protein (GFP) to determine the exact expression of Anxa2-WT, Anxa2-Y23A, and Anxa2-Y23D proteins in the SK-BR-3 cells (Figure 2A). Immunofluorescence studies of Anxa2 also showed an overexpression of Anxa2 in the SK-BR-3 cells, as depicted in Figure 2B. MTT assay results showed that compared with SK-BR-3, control (SK-BR-3 cells expressing empty pCDH lentivirus), and Anxa2-Y23A cells, the Anxa2-WT and Anxa2-Y23D cells exhibited an enhanced ability of duplication (Figure 2C). Anxa2-WT and Anxa2-Y23D enhanced the proliferation of SK-BR-3 cells.

Bottom Line: The monoclonal strains were screened.The monoclonal strains constitutively expressing Anxa2-WT, Anxa2-Y23A, and Anxa2-Y23D were screened.Immunoprecipitation analysis revealed that Anxa2 and Stat3 interacted with each other, and the expression of Stat3 phosphorylation in the nucleus was enhanced by Anxa2-Y23D.

View Article: PubMed Central - PubMed

Affiliation: Tianjin Medical University Cancer Institute and Hospital; Key Laboratory of Cancer Prevention and Therapy, Tianjin; Key Laboratory of Breast Cancer Prevention and Therapy, Ministry of Education, Tianjin 300060, China.

ABSTRACT

Objective: To investigate the role of tyrosine 23 (Tyr23) phosphorylation of Annexin A2 (Anxa2) in regulating the proliferation and invasion of human breast cancer SK-BR-3 cells.

Methods: A panel of lentivirus plasmids expressing Anxa2-wide type (Ana2-WT), Anxa2-Y23A, and Anxa2-Y23D was generated and infected with SK-BR-3 cells. The monoclonal strains were screened. The expression of Anxa2-WT, Anxa2-Y23A, and Anxa2-Y23D was determined by Western blot analysis. The ability of the cells to proliferate was detected through an MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] test. Boyden chamber assays were employed to examine migration and invasion abilities. The interaction between Anxa2 and Stat3 was analyzed by immunoprecipitation analyses. Nucleoprotein and cytosolic protein were extracted from SK-BR-3, Anxa2-WT, Anxa2-Y23A, and Anxa2-Y23D cells to analyze the expression and localization of Stat3 phosphorylation.

Results: The monoclonal strains constitutively expressing Anxa2-WT, Anxa2-Y23A, and Anxa2-Y23D were screened. Both Anxa2-WT and Anxa2-Y23D enhanced the proliferation, migration and invasion abilities of SK-BR-3 cells (P<0.05). Immunoprecipitation analysis revealed that Anxa2 and Stat3 interacted with each other, and the expression of Stat3 phosphorylation in the nucleus was enhanced by Anxa2-Y23D.

Conclusions: Tyr23 phosphorylation of Anxa2 promotes the proliferation and invasion of human breast cancer SK-BR-3 cells and the phosphorylation of Stat3 in the nucleus.

No MeSH data available.


Related in: MedlinePlus