Limits...
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

Co-immunoprecipitation of Anxa2 and Stat3 in SK-BR-3 cells. Immunoprecipitation (IP) and immunoblotting (IB). A and B: The SK-BR-3 cell lysate was immunoprecipitated with the indicated antibodies and immunoblotted with the corresponding antibodies.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3643677&req=5

f4: Co-immunoprecipitation of Anxa2 and Stat3 in SK-BR-3 cells. Immunoprecipitation (IP) and immunoblotting (IB). A and B: The SK-BR-3 cell lysate was immunoprecipitated with the indicated antibodies and immunoblotted with the corresponding antibodies.

Mentions: Co-immunoprecipitation was carried out to study the interaction between Anxa2 and the signal transducer and activator of transcription-3 (Stat3). The SK-BR-3 whole-cell lysates, and protein G-Sepharose beads, and antibodies against Anxa2 and Stat3 were employed to carry out the experiment. Figure 4 shows that the Anxa2 protein was pulled down by the anti-Stat3 antibody, and Stat3 was co-immunoprecipitated with Anxa2 by an anti-Anxa2 antibody. Thus, Anxa2 and Stat3 interacted with each other.


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)

Co-immunoprecipitation of Anxa2 and Stat3 in SK-BR-3 cells. Immunoprecipitation (IP) and immunoblotting (IB). A and B: The SK-BR-3 cell lysate was immunoprecipitated with the indicated antibodies and immunoblotted with the corresponding antibodies.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f4: Co-immunoprecipitation of Anxa2 and Stat3 in SK-BR-3 cells. Immunoprecipitation (IP) and immunoblotting (IB). A and B: The SK-BR-3 cell lysate was immunoprecipitated with the indicated antibodies and immunoblotted with the corresponding antibodies.
Mentions: Co-immunoprecipitation was carried out to study the interaction between Anxa2 and the signal transducer and activator of transcription-3 (Stat3). The SK-BR-3 whole-cell lysates, and protein G-Sepharose beads, and antibodies against Anxa2 and Stat3 were employed to carry out the experiment. Figure 4 shows that the Anxa2 protein was pulled down by the anti-Stat3 antibody, and Stat3 was co-immunoprecipitated with Anxa2 by an anti-Anxa2 antibody. Thus, Anxa2 and Stat3 interacted with each other.

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