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Contrasting effect of recombinant human erythropoietin on breast cancer cell response to cisplatin induced cytotoxicity.

Trost N, Juvan P, Sersa G, Debeljak N - Radiol Oncol (2012)

Bottom Line: Gene expression analysis of p53-dependent genes and bcl-2 gene family members confirmed differences between long and short-term rHuEpo effects, indicating the most prominent changes in BCL2 and BAD expression.On the other hand, MDA-MB-231 cells are almost irresponsive to long-term rHuEpo, supposedly due to the mutated p53 and ER(+)/PR(-) status.The p53 and ER/PR status may predict tumour response on rHuEpo and cDDP treatment.

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: Human recombinant erythropoietin (rHuEpo) that is used for the treatment of the chemotherapy-induced anaemia in cancer patients was shown to cause detrimental effects on the course of disease due to increased adverse events inflicting patient's survival, potentially related to rHuEpo-induced cancer progression. In this study, we elucidate the effect of rHuEpo administration on breast cancer cell proliferation and gene expression after cisplatin (cDDP) induced cytotoxicity.

Materials and methods: Two breast carcinoma models, MCF-7 and MDA-MB-231 cell lines, were used differing in oestrogen (ER) and progesterone (PR) receptors and p53 status. Cells were cultured with or without rHuEpo for 24 h or 9 weeks and their growth characteristics after cDDP treatment were assessed together with expression of genes involved in the p53-signaling pathway.

Results: Short-term exposure of breast cancer cells to rHuEpo lowers their proliferation and reduces cDDP cytotoxic potency. In contrast, long-term exposure of MCF-7 cells to rHuEpo increases proliferation and predisposes MCF-7 cells to cDDP cytotoxicity, but has no effect on MDA-MB-231 cells. MDA-MB-231 cells show altered level of ERK phosphorylation, indicating involvement of MAPK signalling pathway. Gene expression analysis of p53-dependent genes and bcl-2 gene family members confirmed differences between long and short-term rHuEpo effects, indicating the most prominent changes in BCL2 and BAD expression.

Conclusions: Proliferation and survival characteristics of MCF-7 cells are reversely modulated by the length of the rHuEpo exposure. On the other hand, MDA-MB-231 cells are almost irresponsive to long-term rHuEpo, supposedly due to the mutated p53 and ER(+)/PR(-) status. The p53 and ER/PR status may predict tumour response on rHuEpo and cDDP treatment.

No MeSH data available.


Related in: MedlinePlus

Cell proliferation of short (red line, A and C) and long-term (red line, B and D) rHuEpo treated and non-treated cells (black line) after exposure to cDDP, normalized with the proliferation of control cells that were not exposed to cDDP: (A and B) MCF-7; (C and D) MDA-MB-231 cell line. Asterisk (*) denotes statistical significant differences for Type I error α = 0.05.
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f3-rado-46-03-213: Cell proliferation of short (red line, A and C) and long-term (red line, B and D) rHuEpo treated and non-treated cells (black line) after exposure to cDDP, normalized with the proliferation of control cells that were not exposed to cDDP: (A and B) MCF-7; (C and D) MDA-MB-231 cell line. Asterisk (*) denotes statistical significant differences for Type I error α = 0.05.

Mentions: Expression of ERK, Akt and STAT5 proteins and their phosphorylated forms was determined by western blotting in the cell lysates of MCF-7 and MDA-MB-231 cells after rHuEpo treatment and exposure to cDDP. 24 h rHuEpo treated and 9 weeks pretreated cells were together with non-treated cells seeded on 6-well plates in the concentration of 1×105 cells per well and left in culture for 48 h. 24 h before treatments, cells were switched to serum free medium. To assess rHuEpo effect, cells were treated with 5 or 25 U/mL rHuEpo for 15 minutes (similarly as shown in Figure 1C except that rHuEpo treatment was applied instead of cDDP). After treatment, the culture medium was aspirated and samples were fast frozen in liquid nitrogen. To assess rHuEpo and cDDP interaction, cells were exposed to two different concentrations of cDDP for 4 h: 30 and 60 μM for MCF-7 cell line and 60 and 120 μM for MDA-MB-231 (similarly as in Figure 3 except for a shorter cDDP) and fast frozen in liquid nitrogen after culture medium was aspirated.


Contrasting effect of recombinant human erythropoietin on breast cancer cell response to cisplatin induced cytotoxicity.

Trost N, Juvan P, Sersa G, Debeljak N - Radiol Oncol (2012)

Cell proliferation of short (red line, A and C) and long-term (red line, B and D) rHuEpo treated and non-treated cells (black line) after exposure to cDDP, normalized with the proliferation of control cells that were not exposed to cDDP: (A and B) MCF-7; (C and D) MDA-MB-231 cell line. Asterisk (*) denotes statistical significant differences for Type I error α = 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3472952&req=5

f3-rado-46-03-213: Cell proliferation of short (red line, A and C) and long-term (red line, B and D) rHuEpo treated and non-treated cells (black line) after exposure to cDDP, normalized with the proliferation of control cells that were not exposed to cDDP: (A and B) MCF-7; (C and D) MDA-MB-231 cell line. Asterisk (*) denotes statistical significant differences for Type I error α = 0.05.
Mentions: Expression of ERK, Akt and STAT5 proteins and their phosphorylated forms was determined by western blotting in the cell lysates of MCF-7 and MDA-MB-231 cells after rHuEpo treatment and exposure to cDDP. 24 h rHuEpo treated and 9 weeks pretreated cells were together with non-treated cells seeded on 6-well plates in the concentration of 1×105 cells per well and left in culture for 48 h. 24 h before treatments, cells were switched to serum free medium. To assess rHuEpo effect, cells were treated with 5 or 25 U/mL rHuEpo for 15 minutes (similarly as shown in Figure 1C except that rHuEpo treatment was applied instead of cDDP). After treatment, the culture medium was aspirated and samples were fast frozen in liquid nitrogen. To assess rHuEpo and cDDP interaction, cells were exposed to two different concentrations of cDDP for 4 h: 30 and 60 μM for MCF-7 cell line and 60 and 120 μM for MDA-MB-231 (similarly as in Figure 3 except for a shorter cDDP) and fast frozen in liquid nitrogen after culture medium was aspirated.

Bottom Line: Gene expression analysis of p53-dependent genes and bcl-2 gene family members confirmed differences between long and short-term rHuEpo effects, indicating the most prominent changes in BCL2 and BAD expression.On the other hand, MDA-MB-231 cells are almost irresponsive to long-term rHuEpo, supposedly due to the mutated p53 and ER(+)/PR(-) status.The p53 and ER/PR status may predict tumour response on rHuEpo and cDDP treatment.

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: Human recombinant erythropoietin (rHuEpo) that is used for the treatment of the chemotherapy-induced anaemia in cancer patients was shown to cause detrimental effects on the course of disease due to increased adverse events inflicting patient's survival, potentially related to rHuEpo-induced cancer progression. In this study, we elucidate the effect of rHuEpo administration on breast cancer cell proliferation and gene expression after cisplatin (cDDP) induced cytotoxicity.

Materials and methods: Two breast carcinoma models, MCF-7 and MDA-MB-231 cell lines, were used differing in oestrogen (ER) and progesterone (PR) receptors and p53 status. Cells were cultured with or without rHuEpo for 24 h or 9 weeks and their growth characteristics after cDDP treatment were assessed together with expression of genes involved in the p53-signaling pathway.

Results: Short-term exposure of breast cancer cells to rHuEpo lowers their proliferation and reduces cDDP cytotoxic potency. In contrast, long-term exposure of MCF-7 cells to rHuEpo increases proliferation and predisposes MCF-7 cells to cDDP cytotoxicity, but has no effect on MDA-MB-231 cells. MDA-MB-231 cells show altered level of ERK phosphorylation, indicating involvement of MAPK signalling pathway. Gene expression analysis of p53-dependent genes and bcl-2 gene family members confirmed differences between long and short-term rHuEpo effects, indicating the most prominent changes in BCL2 and BAD expression.

Conclusions: Proliferation and survival characteristics of MCF-7 cells are reversely modulated by the length of the rHuEpo exposure. On the other hand, MDA-MB-231 cells are almost irresponsive to long-term rHuEpo, supposedly due to the mutated p53 and ER(+)/PR(-) status. The p53 and ER/PR status may predict tumour response on rHuEpo and cDDP treatment.

No MeSH data available.


Related in: MedlinePlus