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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

Effects of recombinant human EPO on relative EPOR and EPOR-T expression. MCF-7 cells were stimulated with 50 U/ml rHuEPO (short-term, green) or cultured in complete medium in the presence of 5 U/ml of rHuEPO for 10 weeks and stimulated with 50 U/ml rHuEPO (long-term, red). Error bars represent standard deviations (SD) between six replicate samples; asterisk (*) denotes statistical significance for Type 1 error α = 0.05.
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f3-rado-47-04-382: Effects of recombinant human EPO on relative EPOR and EPOR-T expression. MCF-7 cells were stimulated with 50 U/ml rHuEPO (short-term, green) or cultured in complete medium in the presence of 5 U/ml of rHuEPO for 10 weeks and stimulated with 50 U/ml rHuEPO (long-term, red). Error bars represent standard deviations (SD) between six replicate samples; asterisk (*) denotes statistical significance for Type 1 error α = 0.05.

Mentions: To determine the effects of rHuEPO on the expression of its receptor protein variants, mRNA expression levels of EPOR, EPOR-S and EPOR-T genes were analyzed in short (Figure 1A) and long-term (Figure 1C) rHuEPO-treated MCF-7 cells. The expression of EPOR and EPOR-T isoforms at specific time-points was confirmed by qPCR (Figure 3). On the other hand, we were not able to confirm the presence of EPOR-S (data not show). Short-term stimulation of MCF-7 cells with 50 U/ml rHuEPO leads to an increase in EPOR expression, while it has no statistically significant effect on EPOR-T. Interestingly, the addition of 50 U/ml rHuEPO to the long-term pretreated cells (5 U/ml rHuEPO) did not have any additional influence on the expression levels of EPOR and EPOR-T.


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)

Effects of recombinant human EPO on relative EPOR and EPOR-T expression. MCF-7 cells were stimulated with 50 U/ml rHuEPO (short-term, green) or cultured in complete medium in the presence of 5 U/ml of rHuEPO for 10 weeks and stimulated with 50 U/ml rHuEPO (long-term, red). Error bars represent standard deviations (SD) between six replicate samples; 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

f3-rado-47-04-382: Effects of recombinant human EPO on relative EPOR and EPOR-T expression. MCF-7 cells were stimulated with 50 U/ml rHuEPO (short-term, green) or cultured in complete medium in the presence of 5 U/ml of rHuEPO for 10 weeks and stimulated with 50 U/ml rHuEPO (long-term, red). Error bars represent standard deviations (SD) between six replicate samples; asterisk (*) denotes statistical significance for Type 1 error α = 0.05.
Mentions: To determine the effects of rHuEPO on the expression of its receptor protein variants, mRNA expression levels of EPOR, EPOR-S and EPOR-T genes were analyzed in short (Figure 1A) and long-term (Figure 1C) rHuEPO-treated MCF-7 cells. The expression of EPOR and EPOR-T isoforms at specific time-points was confirmed by qPCR (Figure 3). On the other hand, we were not able to confirm the presence of EPOR-S (data not show). Short-term stimulation of MCF-7 cells with 50 U/ml rHuEPO leads to an increase in EPOR expression, while it has no statistically significant effect on EPOR-T. Interestingly, the addition of 50 U/ml rHuEPO to the long-term pretreated cells (5 U/ml rHuEPO) did not have any additional influence on the expression levels of EPOR and EPOR-T.

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