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SIP1/NHERF2 enhances estrogen receptor alpha transactivation in breast cancer cells.

Meneses-Morales I, Tecalco-Cruz AC, Barrios-García T, Gómez-Romero V, Trujillo-González I, Reyes-Carmona S, García-Zepeda E, Méndez-Enríquez E, Cervantes-Roldán R, Pérez-Sánchez V, Recillas-Targa F, Mohar-Betancourt A, León-Del-Río A - Nucleic Acids Res. (2014)

Bottom Line: In this study, we have identified the protein Na+/H+ Exchanger RegulatoryFactor 2 (NHERF2) as an ERα-associated coactivator that interacts predominantly with the AF-1 domain of the nuclear receptor.We analyzed the expression of NHERF2 in breast cancer tumors finding a 2- to 17-fold increase in its mRNA levels in 50% of the tumor samples compared to normal breast tissue.These results indicate that NHERF2 is a coactivator of ERα that may participate in the development of estrogen-dependent breast cancer tumors.

View Article: PubMed Central - PubMed

Affiliation: Programa de Investigación de Cáncer de Mama, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, México D.F. 04510, Mexico Departamento de Biología Molecular y Biotecnología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, México D.F. 04510, Mexico.

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NHERF2 enhances proliferation and tumorigenic potential of MCF7 cells. The effect of NHERF2 overexpression on MCF7 cells proliferation was analyzed using the CFSE assay as described in Materials and Methods. Control MCF7 and FLAG-NHERF2-MCF7 cells were analyzed every 24 h after CFSE labeling by FACS. Representative histograms showing the fluorescence intensity distribution of MCF7 and FLAG-NHERF2-MCF7 transfected cells at 24 h (A) and 48 h (B) after CFSE labeling compared with MCF7 control (time 0) cells. (C) Representative histogram displaying CFSE fold reduction of FLAG-NHERF2-MCF7 cells (gray bars) compared to control MCF7 cells (white bars) at 24 and 48 h after CFSE labeling. (D) NHERF2 overexpression increases the tumorigenic potential of MCF7 cells in a nu/nu mouse model. Two groups of 8 nu/nu mice were injected with 3×106 control MCF7 cells or 3×106 FLAG-NHERF2-MCF7 cells as described in Materials and Methods. After 30 days the animals were sacrificed and the tumors were isolated and weighted. Tumor weight is represented as mean ± S.E.
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Figure 5: NHERF2 enhances proliferation and tumorigenic potential of MCF7 cells. The effect of NHERF2 overexpression on MCF7 cells proliferation was analyzed using the CFSE assay as described in Materials and Methods. Control MCF7 and FLAG-NHERF2-MCF7 cells were analyzed every 24 h after CFSE labeling by FACS. Representative histograms showing the fluorescence intensity distribution of MCF7 and FLAG-NHERF2-MCF7 transfected cells at 24 h (A) and 48 h (B) after CFSE labeling compared with MCF7 control (time 0) cells. (C) Representative histogram displaying CFSE fold reduction of FLAG-NHERF2-MCF7 cells (gray bars) compared to control MCF7 cells (white bars) at 24 and 48 h after CFSE labeling. (D) NHERF2 overexpression increases the tumorigenic potential of MCF7 cells in a nu/nu mouse model. Two groups of 8 nu/nu mice were injected with 3×106 control MCF7 cells or 3×106 FLAG-NHERF2-MCF7 cells as described in Materials and Methods. After 30 days the animals were sacrificed and the tumors were isolated and weighted. Tumor weight is represented as mean ± S.E.

Mentions: In human breast cancer cells the activity of ERα is associated with cell proliferation. To test whether NHERF2 overexpressing cells proliferate at a different rate than control MCF7 cells, we used a cell tracing assay in which cells treated with CFSE are allowed to divide in culture for 24 h or 48 h. After labeling, all cells were uniformly stained with CFSE. However, after 24 h and 48 h in culture NHERF2-MCF7 cells retained less CFSE than control MCF7 cells (Figure 5A–C), indicating a slower rate of cell division by the control cells.


SIP1/NHERF2 enhances estrogen receptor alpha transactivation in breast cancer cells.

Meneses-Morales I, Tecalco-Cruz AC, Barrios-García T, Gómez-Romero V, Trujillo-González I, Reyes-Carmona S, García-Zepeda E, Méndez-Enríquez E, Cervantes-Roldán R, Pérez-Sánchez V, Recillas-Targa F, Mohar-Betancourt A, León-Del-Río A - Nucleic Acids Res. (2014)

NHERF2 enhances proliferation and tumorigenic potential of MCF7 cells. The effect of NHERF2 overexpression on MCF7 cells proliferation was analyzed using the CFSE assay as described in Materials and Methods. Control MCF7 and FLAG-NHERF2-MCF7 cells were analyzed every 24 h after CFSE labeling by FACS. Representative histograms showing the fluorescence intensity distribution of MCF7 and FLAG-NHERF2-MCF7 transfected cells at 24 h (A) and 48 h (B) after CFSE labeling compared with MCF7 control (time 0) cells. (C) Representative histogram displaying CFSE fold reduction of FLAG-NHERF2-MCF7 cells (gray bars) compared to control MCF7 cells (white bars) at 24 and 48 h after CFSE labeling. (D) NHERF2 overexpression increases the tumorigenic potential of MCF7 cells in a nu/nu mouse model. Two groups of 8 nu/nu mice were injected with 3×106 control MCF7 cells or 3×106 FLAG-NHERF2-MCF7 cells as described in Materials and Methods. After 30 days the animals were sacrificed and the tumors were isolated and weighted. Tumor weight is represented as mean ± S.E.
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Related In: Results  -  Collection

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Figure 5: NHERF2 enhances proliferation and tumorigenic potential of MCF7 cells. The effect of NHERF2 overexpression on MCF7 cells proliferation was analyzed using the CFSE assay as described in Materials and Methods. Control MCF7 and FLAG-NHERF2-MCF7 cells were analyzed every 24 h after CFSE labeling by FACS. Representative histograms showing the fluorescence intensity distribution of MCF7 and FLAG-NHERF2-MCF7 transfected cells at 24 h (A) and 48 h (B) after CFSE labeling compared with MCF7 control (time 0) cells. (C) Representative histogram displaying CFSE fold reduction of FLAG-NHERF2-MCF7 cells (gray bars) compared to control MCF7 cells (white bars) at 24 and 48 h after CFSE labeling. (D) NHERF2 overexpression increases the tumorigenic potential of MCF7 cells in a nu/nu mouse model. Two groups of 8 nu/nu mice were injected with 3×106 control MCF7 cells or 3×106 FLAG-NHERF2-MCF7 cells as described in Materials and Methods. After 30 days the animals were sacrificed and the tumors were isolated and weighted. Tumor weight is represented as mean ± S.E.
Mentions: In human breast cancer cells the activity of ERα is associated with cell proliferation. To test whether NHERF2 overexpressing cells proliferate at a different rate than control MCF7 cells, we used a cell tracing assay in which cells treated with CFSE are allowed to divide in culture for 24 h or 48 h. After labeling, all cells were uniformly stained with CFSE. However, after 24 h and 48 h in culture NHERF2-MCF7 cells retained less CFSE than control MCF7 cells (Figure 5A–C), indicating a slower rate of cell division by the control cells.

Bottom Line: In this study, we have identified the protein Na+/H+ Exchanger RegulatoryFactor 2 (NHERF2) as an ERα-associated coactivator that interacts predominantly with the AF-1 domain of the nuclear receptor.We analyzed the expression of NHERF2 in breast cancer tumors finding a 2- to 17-fold increase in its mRNA levels in 50% of the tumor samples compared to normal breast tissue.These results indicate that NHERF2 is a coactivator of ERα that may participate in the development of estrogen-dependent breast cancer tumors.

View Article: PubMed Central - PubMed

Affiliation: Programa de Investigación de Cáncer de Mama, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, México D.F. 04510, Mexico Departamento de Biología Molecular y Biotecnología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, México D.F. 04510, Mexico.

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Related in: MedlinePlus