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Identification of the Interaction between P-Glycoprotein and Anxa2 in Multidrug-resistant Human Breast Cancer Cells.

Zhang HC, Zhang F, Wu B, Han JH, Ji W, Zhou Y, Niu RF - Cancer Biol Med (2012)

Bottom Line: The expression of P-gp was detected by Western blot.There was a close interaction between Anxa2 and P-gp.The interaction of Anxa2 with P-pg may play an important role in the enhanced invasiveness of MDR human breast cancer cells.

View Article: PubMed Central - PubMed

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

ABSTRACT

Objective: To explore the interaction of Anxa2 with P-Glycoprotein (P-gp) in the migration and invasion of the multidrug-resistant (MDR) human breast cancer cell line MCF-7/ADR.

Methods: A pair of short hairpin RNA (shRNA) targeting P-gp was transfected into MCF-7/ADR cells, and monoclonal cell strains were screened. The expression of P-gp was detected by Western blot. Transwell chambers were used to observe the cell migration capacity and invasion ability. The interaction between P-gp and Anxa2 was examined by immunoprecipitation and immunofluorescence confocal microscopy analyses.

Results: P-gp expression was significantly knocked down, and there were notable decreasing trends in the migration and invasion capability of MDR breast cancer cells (P<0.05). There was a close interaction between Anxa2 and P-gp.

Conclusions: MCF-7/ADR is an MDR human breast cancer cell line with high migration and invasion abilities. The knockdown of P-gp notably impaired the migration and invasion abilities of the tumor cells. The interaction of Anxa2 with P-pg may play an important role in the enhanced invasiveness of MDR human breast cancer cells.

No MeSH data available.


Related in: MedlinePlus

Co-immunoprecipitation of Anxa2 and P-gp in MCF-7/ADR cells. Immunoprecipitation (IP) and immunoblotting (IB). The MCF-7/ADR cell lysate was immunoprecipitated with the indicated antibodies and immunoblotted with corresponding antibodies.
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f3: Co-immunoprecipitation of Anxa2 and P-gp in MCF-7/ADR cells. Immunoprecipitation (IP) and immunoblotting (IB). The MCF-7/ADR cell lysate was immunoprecipitated with the indicated antibodies and immunoblotted with corresponding antibodies.

Mentions: Co-immunoprecipitation was carried out in MCF-7/ADR whole cell lysates with protein G-Sepharose beads and antibodies against Anxa2 and P-gp. As shown in Figure 3A, Anxa2 protein was pulled down by anti-P-gp antibody, and P-gp was also co-immunoprecipitated with Anxa2 by an anti-Anxa2 antibody (Figure 3B). The cellular localization of the 2 proteins was also analyzed by confocal microscopy. The double immunofluorescent analysis showed that the expression of P-gp (green) in MCF-7/ADR cells merged with Anxa2 (red), suggesting their physical association with MCF-7/ADR cells (Figure 4A). A wound-healing assay obtained similar results. As indicated in Figure 4B, P-gp and Anxa2 were co-expressed at the edge of the “wound” of MCF-7/ADR cells.


Identification of the Interaction between P-Glycoprotein and Anxa2 in Multidrug-resistant Human Breast Cancer Cells.

Zhang HC, Zhang F, Wu B, Han JH, Ji W, Zhou Y, Niu RF - Cancer Biol Med (2012)

Co-immunoprecipitation of Anxa2 and P-gp in MCF-7/ADR cells. Immunoprecipitation (IP) and immunoblotting (IB). The MCF-7/ADR cell lysate was immunoprecipitated with the indicated antibodies and immunoblotted with corresponding antibodies.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3643660&req=5

f3: Co-immunoprecipitation of Anxa2 and P-gp in MCF-7/ADR cells. Immunoprecipitation (IP) and immunoblotting (IB). The MCF-7/ADR cell lysate was immunoprecipitated with the indicated antibodies and immunoblotted with corresponding antibodies.
Mentions: Co-immunoprecipitation was carried out in MCF-7/ADR whole cell lysates with protein G-Sepharose beads and antibodies against Anxa2 and P-gp. As shown in Figure 3A, Anxa2 protein was pulled down by anti-P-gp antibody, and P-gp was also co-immunoprecipitated with Anxa2 by an anti-Anxa2 antibody (Figure 3B). The cellular localization of the 2 proteins was also analyzed by confocal microscopy. The double immunofluorescent analysis showed that the expression of P-gp (green) in MCF-7/ADR cells merged with Anxa2 (red), suggesting their physical association with MCF-7/ADR cells (Figure 4A). A wound-healing assay obtained similar results. As indicated in Figure 4B, P-gp and Anxa2 were co-expressed at the edge of the “wound” of MCF-7/ADR cells.

Bottom Line: The expression of P-gp was detected by Western blot.There was a close interaction between Anxa2 and P-gp.The interaction of Anxa2 with P-pg may play an important role in the enhanced invasiveness of MDR human breast cancer cells.

View Article: PubMed Central - PubMed

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

ABSTRACT

Objective: To explore the interaction of Anxa2 with P-Glycoprotein (P-gp) in the migration and invasion of the multidrug-resistant (MDR) human breast cancer cell line MCF-7/ADR.

Methods: A pair of short hairpin RNA (shRNA) targeting P-gp was transfected into MCF-7/ADR cells, and monoclonal cell strains were screened. The expression of P-gp was detected by Western blot. Transwell chambers were used to observe the cell migration capacity and invasion ability. The interaction between P-gp and Anxa2 was examined by immunoprecipitation and immunofluorescence confocal microscopy analyses.

Results: P-gp expression was significantly knocked down, and there were notable decreasing trends in the migration and invasion capability of MDR breast cancer cells (P<0.05). There was a close interaction between Anxa2 and P-gp.

Conclusions: MCF-7/ADR is an MDR human breast cancer cell line with high migration and invasion abilities. The knockdown of P-gp notably impaired the migration and invasion abilities of the tumor cells. The interaction of Anxa2 with P-pg may play an important role in the enhanced invasiveness of MDR human breast cancer cells.

No MeSH data available.


Related in: MedlinePlus