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Recombinant human erythropoietin alters gene expression and stimulates proliferation of MCF-7 breast cancer cells.

Trost N, Stepisnik T, Berne S, Pucer A, Petan T, Komel R, Debeljak N - Radiol Oncol (2013)

Bottom Line: Three different isoforms of erythropoietin receptor (EPOR) have been reported, the soluble (EPOR-S) and truncated (EPOR-T) forms acting antagonistically to the functional EPOR.Gene expression analysis showed activation of EGR1 and FOS, confirming the functionality of EPOR. rHuEPO treatment also slightly increased the expression of the functional EPOR isoform, which, however, persisted throughout the 10 weeks of treatment.The expression levels of EPOR-T were not influenced.

View Article: PubMed Central - PubMed

Affiliation: Center for Functional Genomics and Bio-chips, Institute of Biochemistry, Faculty of Medicine, University of Ljubljana, Slovenia.

ABSTRACT

Background: Functional erythropoietin (EPO) signaling is not specific only to erythroid lineages and has been confirmed in several solid tumors, including breast. Three different isoforms of erythropoietin receptor (EPOR) have been reported, the soluble (EPOR-S) and truncated (EPOR-T) forms acting antagonistically to the functional EPOR. In this study, we investigated the effect of human recombinant erythropoietin (rHuEPO) on cell proliferation, early gene response and the expression of EPOR isoforms in the MCF-7 breast cancer cell line.

Materials and methods: The MCF-7 cells were cultured with or without rHuEPO for 72 h or 10 weeks and assessed for their growth characteristics, expression of early response genes and different EPOR isoforms. The expression profile of EPOR and EPOR-T was determined in a range of breast cancer cell lines and compared with their invasive properties.

Results: MCF-7 cell proliferation after rHuEPO treatment was dependent on the time of treatment and the concentration used. High rHuEPO concentrations (40 U/ml) stimulated cell proliferation independently of a preceding long-term exposure of MCF-7 cells to rHuEPO, while lower concentrations increased MCF-7 proliferation only after 10 weeks of treatment. Gene expression analysis showed activation of EGR1 and FOS, confirming the functionality of EPOR. rHuEPO treatment also slightly increased the expression of the functional EPOR isoform, which, however, persisted throughout the 10 weeks of treatment. The expression levels of EPOR-T were not influenced. There were no correlations between EPOR expression and the invasiveness of MCF-7, MDA-MB-231, Hs578T, Hs578Bst, SKBR3, T-47D and MCF-10A cell lines.

Conclusions: rHuEPO modulates MCF-7 cell proliferation in time- and concentration-dependent manner. We confirmed EGR1, FOS and EPOR as transcription targets of the EPO-EPOR signaling loop, but could not correlate the expression of different EPOR isoforms with the invasiveness of breast cancer cell lines.

No MeSH data available.


Related in: MedlinePlus

Differential effects of recombinant human EPO on MCF-7 cell proliferation (A) MCF-7 cells were cultured in complete medium in the presence of indicated concentrations of rHuEPO (short-term treated) (B) MCF-7 cells were cultured in complete medium in the presence of 5 U/ml of rHuEPO for 10 weeks (long-term pretreated cells), EPO was added to the pretreated cells at indicated concentrations. Asterisk (*) denotes statistical significance for Type 1 error α = 0.05.
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f2-rado-47-04-382: Differential effects of recombinant human EPO on MCF-7 cell proliferation (A) MCF-7 cells were cultured in complete medium in the presence of indicated concentrations of rHuEPO (short-term treated) (B) MCF-7 cells were cultured in complete medium in the presence of 5 U/ml of rHuEPO for 10 weeks (long-term pretreated cells), EPO was added to the pretreated cells at indicated concentrations. Asterisk (*) denotes statistical significance for Type 1 error α = 0.05.

Mentions: MCF-7 cells were stimulated with rHuEPO (0, 5, 40 U/ml) and assessed for proliferation using the MTT assay. We found that MCF-7 cell proliferation is dependent on the concentration of rHuEPO used and the time of the treatment (Figure 2). Treatments with 40 U/ml rHuEPO led to increased MCF-7 cell proliferation independently of the length of cell exposure to rHuEPO. On the other hand, 5 U/ml rHuEPO affects MCF-7 cell proliferation in a time dependent manner; cell proliferation was reduced during a short-term treatment (Figure 2A), but was higher when rHuEPO was added to long-term rHuEPO-pretreated cells (Figure 2B).


Recombinant human erythropoietin alters gene expression and stimulates proliferation of MCF-7 breast cancer cells.

Trost N, Stepisnik T, Berne S, Pucer A, Petan T, Komel R, Debeljak N - Radiol Oncol (2013)

Differential effects of recombinant human EPO on MCF-7 cell proliferation (A) MCF-7 cells were cultured in complete medium in the presence of indicated concentrations of rHuEPO (short-term treated) (B) MCF-7 cells were cultured in complete medium in the presence of 5 U/ml of rHuEPO for 10 weeks (long-term pretreated cells), EPO was added to the pretreated cells at indicated concentrations. Asterisk (*) denotes statistical significance for Type 1 error α = 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3814284&req=5

f2-rado-47-04-382: Differential effects of recombinant human EPO on MCF-7 cell proliferation (A) MCF-7 cells were cultured in complete medium in the presence of indicated concentrations of rHuEPO (short-term treated) (B) MCF-7 cells were cultured in complete medium in the presence of 5 U/ml of rHuEPO for 10 weeks (long-term pretreated cells), EPO was added to the pretreated cells at indicated concentrations. Asterisk (*) denotes statistical significance for Type 1 error α = 0.05.
Mentions: MCF-7 cells were stimulated with rHuEPO (0, 5, 40 U/ml) and assessed for proliferation using the MTT assay. We found that MCF-7 cell proliferation is dependent on the concentration of rHuEPO used and the time of the treatment (Figure 2). Treatments with 40 U/ml rHuEPO led to increased MCF-7 cell proliferation independently of the length of cell exposure to rHuEPO. On the other hand, 5 U/ml rHuEPO affects MCF-7 cell proliferation in a time dependent manner; cell proliferation was reduced during a short-term treatment (Figure 2A), but was higher when rHuEPO was added to long-term rHuEPO-pretreated cells (Figure 2B).

Bottom Line: Three different isoforms of erythropoietin receptor (EPOR) have been reported, the soluble (EPOR-S) and truncated (EPOR-T) forms acting antagonistically to the functional EPOR.Gene expression analysis showed activation of EGR1 and FOS, confirming the functionality of EPOR. rHuEPO treatment also slightly increased the expression of the functional EPOR isoform, which, however, persisted throughout the 10 weeks of treatment.The expression levels of EPOR-T were not influenced.

View Article: PubMed Central - PubMed

Affiliation: Center for Functional Genomics and Bio-chips, Institute of Biochemistry, Faculty of Medicine, University of Ljubljana, Slovenia.

ABSTRACT

Background: Functional erythropoietin (EPO) signaling is not specific only to erythroid lineages and has been confirmed in several solid tumors, including breast. Three different isoforms of erythropoietin receptor (EPOR) have been reported, the soluble (EPOR-S) and truncated (EPOR-T) forms acting antagonistically to the functional EPOR. In this study, we investigated the effect of human recombinant erythropoietin (rHuEPO) on cell proliferation, early gene response and the expression of EPOR isoforms in the MCF-7 breast cancer cell line.

Materials and methods: The MCF-7 cells were cultured with or without rHuEPO for 72 h or 10 weeks and assessed for their growth characteristics, expression of early response genes and different EPOR isoforms. The expression profile of EPOR and EPOR-T was determined in a range of breast cancer cell lines and compared with their invasive properties.

Results: MCF-7 cell proliferation after rHuEPO treatment was dependent on the time of treatment and the concentration used. High rHuEPO concentrations (40 U/ml) stimulated cell proliferation independently of a preceding long-term exposure of MCF-7 cells to rHuEPO, while lower concentrations increased MCF-7 proliferation only after 10 weeks of treatment. Gene expression analysis showed activation of EGR1 and FOS, confirming the functionality of EPOR. rHuEPO treatment also slightly increased the expression of the functional EPOR isoform, which, however, persisted throughout the 10 weeks of treatment. The expression levels of EPOR-T were not influenced. There were no correlations between EPOR expression and the invasiveness of MCF-7, MDA-MB-231, Hs578T, Hs578Bst, SKBR3, T-47D and MCF-10A cell lines.

Conclusions: rHuEPO modulates MCF-7 cell proliferation in time- and concentration-dependent manner. We confirmed EGR1, FOS and EPOR as transcription targets of the EPO-EPOR signaling loop, but could not correlate the expression of different EPOR isoforms with the invasiveness of breast cancer cell lines.

No MeSH data available.


Related in: MedlinePlus