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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-Y23D increased the expression of phosphorylation Stat3 in nucleus. A: Phosphorylation Anxa2 expression increased as the exposure time was lengthened, whereas the expression of other proteins did not show any significant changes. B: Western blot analysis of the expression of Stat3 and phosphorylation Stat3, Anxa2 and Phosphorylation Anxa2 in SK-BR-3, Anxa2-WT, Anxa2-Y23A and Anxa2-Y23D cells, with no significant differences. C: Nucleoprotein and cytosolic protein were extracted from SK-BR-3, Anxa2-WT, Anxa2-Y23A, and Anxa2-Y23D cells. Western blot analysis showed that the expression of phosphorylation Stat3 was notably enhanced in Anxa2-Y23D cells.
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f5: Anxa2-Y23D increased the expression of phosphorylation Stat3 in nucleus. A: Phosphorylation Anxa2 expression increased as the exposure time was lengthened, whereas the expression of other proteins did not show any significant changes. B: Western blot analysis of the expression of Stat3 and phosphorylation Stat3, Anxa2 and Phosphorylation Anxa2 in SK-BR-3, Anxa2-WT, Anxa2-Y23A and Anxa2-Y23D cells, with no significant differences. C: Nucleoprotein and cytosolic protein were extracted from SK-BR-3, Anxa2-WT, Anxa2-Y23A, and Anxa2-Y23D cells. Western blot analysis showed that the expression of phosphorylation Stat3 was notably enhanced in Anxa2-Y23D cells.

Mentions: The interaction between Anxa2 and Stat3 in the SK-BR-3 cells in vitro led to the hypothesis that Stat3 and the phosphorylation Stat3 (P-Stat3) may be up-regulated in Anxa2-WT or Anxa2-Y23D cells. Western blot results suggested no significant differences in the expression of Stat3 and phosphorylation Stat3 (P-Stat3) in both SK-BR-3 and Anxa2-WT cells among different groups exposed to EGF for 0, 5, and 10 min (Figure 5A). However, the expression of phosphorylation Anxa2 (P-Anxa2) was increased when the cells were stimulated by EGF for 10 min (Figure 5A). The expression of Stat3 and phosphorylation Stat3, Anxa2 and phosphorylation Anxa2 also exhibited no significant differences among the SK-BR-3, Anxa2-WT, Anxa2-Y23A, and Anxa2-Y23D cells (Figure 5B). Anxa3-WT and Anxa2-Y23D clearly had no effect on the expression of Stat3 and phosphorylation Stat3 in the total cell lysate. A panel of nucleoproteins and cytosolic protein were extracted from SK-BR-3, Anxa2-WT, Anxa2-Y23A, and Anxa2-Y23D cells, and Western blot analysis showed that phosphorylation Stat3 was notably localized in the nucleus of Anxa2-Y23D cells (Figure 5C).


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-Y23D increased the expression of phosphorylation Stat3 in nucleus. A: Phosphorylation Anxa2 expression increased as the exposure time was lengthened, whereas the expression of other proteins did not show any significant changes. B: Western blot analysis of the expression of Stat3 and phosphorylation Stat3, Anxa2 and Phosphorylation Anxa2 in SK-BR-3, Anxa2-WT, Anxa2-Y23A and Anxa2-Y23D cells, with no significant differences. C: Nucleoprotein and cytosolic protein were extracted from SK-BR-3, Anxa2-WT, Anxa2-Y23A, and Anxa2-Y23D cells. Western blot analysis showed that the expression of phosphorylation Stat3 was notably enhanced in Anxa2-Y23D cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f5: Anxa2-Y23D increased the expression of phosphorylation Stat3 in nucleus. A: Phosphorylation Anxa2 expression increased as the exposure time was lengthened, whereas the expression of other proteins did not show any significant changes. B: Western blot analysis of the expression of Stat3 and phosphorylation Stat3, Anxa2 and Phosphorylation Anxa2 in SK-BR-3, Anxa2-WT, Anxa2-Y23A and Anxa2-Y23D cells, with no significant differences. C: Nucleoprotein and cytosolic protein were extracted from SK-BR-3, Anxa2-WT, Anxa2-Y23A, and Anxa2-Y23D cells. Western blot analysis showed that the expression of phosphorylation Stat3 was notably enhanced in Anxa2-Y23D cells.
Mentions: The interaction between Anxa2 and Stat3 in the SK-BR-3 cells in vitro led to the hypothesis that Stat3 and the phosphorylation Stat3 (P-Stat3) may be up-regulated in Anxa2-WT or Anxa2-Y23D cells. Western blot results suggested no significant differences in the expression of Stat3 and phosphorylation Stat3 (P-Stat3) in both SK-BR-3 and Anxa2-WT cells among different groups exposed to EGF for 0, 5, and 10 min (Figure 5A). However, the expression of phosphorylation Anxa2 (P-Anxa2) was increased when the cells were stimulated by EGF for 10 min (Figure 5A). The expression of Stat3 and phosphorylation Stat3, Anxa2 and phosphorylation Anxa2 also exhibited no significant differences among the SK-BR-3, Anxa2-WT, Anxa2-Y23A, and Anxa2-Y23D cells (Figure 5B). Anxa3-WT and Anxa2-Y23D clearly had no effect on the expression of Stat3 and phosphorylation Stat3 in the total cell lysate. A panel of nucleoproteins and cytosolic protein were extracted from SK-BR-3, Anxa2-WT, Anxa2-Y23A, and Anxa2-Y23D cells, and Western blot analysis showed that phosphorylation Stat3 was notably localized in the nucleus of Anxa2-Y23D cells (Figure 5C).

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