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Assessment of gene expression of intracellular calcium channels, pumps and exchangers with epidermal growth factor-induced epithelial-mesenchymal transition in a breast cancer cell line.

Davis FM, Parsonage MT, Cabot PJ, Parat MO, Thompson EW, Roberts-Thomson SJ, Monteith GR - Cancer Cell Int. (2013)

Bottom Line: These data reveal no significant alterations in mRNA levels of the Golgi calcium pump secretory pathway calcium ATPases (SPCA1 and SPCA2), or the mitochondrial calcium uniporter (MCU) or Na+/Ca2+ exchanger (NCLX).However, EGF-induced EMT was associated with significant alterations in mRNA levels of specific ER calcium channels and pumps, including (sarco)-endoplasmic reticulum calcium ATPases (SERCAs), and inositol 1,4,5-trisphosphate receptor (IP3R) and ryanodine receptor (RYR) calcium channel isoforms.The most prominent change in gene expression between the epithelial and mesenchymal-like states was RYR2, which was enriched 45-fold in EGF-treated MDA-MB-468 cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: School of Pharmacy, The University of Queensland, Brisbane, QLD 4072, Australia. gregm@uq.edu.au.

ABSTRACT

Background: Epithelial-mesenchymal transition (EMT) is a process implicated in cancer metastasis that involves the conversion of epithelial cells to a more mesenchymal and invasive cell phenotype. In breast cancer cells EMT is associated with altered store-operated calcium influx and changes in calcium signalling mediated by activation of cell surface purinergic receptors. In this study, we investigated whether MDA-MB-468 breast cancer cells induced to undergo EMT exhibit changes in mRNA levels of calcium channels, pumps and exchangers located on intracellular calcium storing organelles, including the Golgi, mitochondria and endoplasmic reticulum (ER).

Methods: Epidermal growth factor (EGF) was used to induce EMT in MDA-MB-468 breast cancer cells. Serum-deprived cells were treated with EGF (50 ng/mL) for 12 h and gene expression was assessed using quantitative RT-PCR.

Results and conclusions: These data reveal no significant alterations in mRNA levels of the Golgi calcium pump secretory pathway calcium ATPases (SPCA1 and SPCA2), or the mitochondrial calcium uniporter (MCU) or Na+/Ca2+ exchanger (NCLX). However, EGF-induced EMT was associated with significant alterations in mRNA levels of specific ER calcium channels and pumps, including (sarco)-endoplasmic reticulum calcium ATPases (SERCAs), and inositol 1,4,5-trisphosphate receptor (IP3R) and ryanodine receptor (RYR) calcium channel isoforms. The most prominent change in gene expression between the epithelial and mesenchymal-like states was RYR2, which was enriched 45-fold in EGF-treated MDA-MB-468 cells. These findings indicate that EGF-induced EMT in breast cancer cells may be associated with major alterations in ER calcium homeostasis.

No MeSH data available.


Related in: MedlinePlus

EGF-induced EMT in MDA-MB-468 breast cancer cells. MDA-MB-468 breast cancer cells were serum starved and treated with EGF (50 ng/mL) as indicated to induce EMT. A) Twist and B) vimentin mRNA expression were assessed using real time RT-PCR at 3, 6 and 12 h following EGF treatment. Bar graphs show mean ± S.D. for nine individual wells from three independent experiments. The effect of EGF at each time point was assessed using two-way ANOVA with Bonferroni’s multiple comparisons post-tests. * P < 0.05.
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Figure 1: EGF-induced EMT in MDA-MB-468 breast cancer cells. MDA-MB-468 breast cancer cells were serum starved and treated with EGF (50 ng/mL) as indicated to induce EMT. A) Twist and B) vimentin mRNA expression were assessed using real time RT-PCR at 3, 6 and 12 h following EGF treatment. Bar graphs show mean ± S.D. for nine individual wells from three independent experiments. The effect of EGF at each time point was assessed using two-way ANOVA with Bonferroni’s multiple comparisons post-tests. * P < 0.05.

Mentions: MDA-MB-468 breast cancer cells treated with EGF (50 ng/mL) for 3, 6 or 12 h showed a significant increase in mRNA levels of Twist (Figure 1A), a nuclear transcription factor and marker of EMT [22]. This early EMT event was followed by an increase in vimentin mRNA at 12 h post-EGF treatment (Figure 1B). These findings were consistent with previous studies using this model of EGF-induced EMT [5]–[8]. We assessed gene expression of calcium channels, pumps and exchangers located on the Golgi, mitochondria and ER with EMT after treatment with EGF for 12 h.


Assessment of gene expression of intracellular calcium channels, pumps and exchangers with epidermal growth factor-induced epithelial-mesenchymal transition in a breast cancer cell line.

Davis FM, Parsonage MT, Cabot PJ, Parat MO, Thompson EW, Roberts-Thomson SJ, Monteith GR - Cancer Cell Int. (2013)

EGF-induced EMT in MDA-MB-468 breast cancer cells. MDA-MB-468 breast cancer cells were serum starved and treated with EGF (50 ng/mL) as indicated to induce EMT. A) Twist and B) vimentin mRNA expression were assessed using real time RT-PCR at 3, 6 and 12 h following EGF treatment. Bar graphs show mean ± S.D. for nine individual wells from three independent experiments. The effect of EGF at each time point was assessed using two-way ANOVA with Bonferroni’s multiple comparisons post-tests. * P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3733826&req=5

Figure 1: EGF-induced EMT in MDA-MB-468 breast cancer cells. MDA-MB-468 breast cancer cells were serum starved and treated with EGF (50 ng/mL) as indicated to induce EMT. A) Twist and B) vimentin mRNA expression were assessed using real time RT-PCR at 3, 6 and 12 h following EGF treatment. Bar graphs show mean ± S.D. for nine individual wells from three independent experiments. The effect of EGF at each time point was assessed using two-way ANOVA with Bonferroni’s multiple comparisons post-tests. * P < 0.05.
Mentions: MDA-MB-468 breast cancer cells treated with EGF (50 ng/mL) for 3, 6 or 12 h showed a significant increase in mRNA levels of Twist (Figure 1A), a nuclear transcription factor and marker of EMT [22]. This early EMT event was followed by an increase in vimentin mRNA at 12 h post-EGF treatment (Figure 1B). These findings were consistent with previous studies using this model of EGF-induced EMT [5]–[8]. We assessed gene expression of calcium channels, pumps and exchangers located on the Golgi, mitochondria and ER with EMT after treatment with EGF for 12 h.

Bottom Line: These data reveal no significant alterations in mRNA levels of the Golgi calcium pump secretory pathway calcium ATPases (SPCA1 and SPCA2), or the mitochondrial calcium uniporter (MCU) or Na+/Ca2+ exchanger (NCLX).However, EGF-induced EMT was associated with significant alterations in mRNA levels of specific ER calcium channels and pumps, including (sarco)-endoplasmic reticulum calcium ATPases (SERCAs), and inositol 1,4,5-trisphosphate receptor (IP3R) and ryanodine receptor (RYR) calcium channel isoforms.The most prominent change in gene expression between the epithelial and mesenchymal-like states was RYR2, which was enriched 45-fold in EGF-treated MDA-MB-468 cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: School of Pharmacy, The University of Queensland, Brisbane, QLD 4072, Australia. gregm@uq.edu.au.

ABSTRACT

Background: Epithelial-mesenchymal transition (EMT) is a process implicated in cancer metastasis that involves the conversion of epithelial cells to a more mesenchymal and invasive cell phenotype. In breast cancer cells EMT is associated with altered store-operated calcium influx and changes in calcium signalling mediated by activation of cell surface purinergic receptors. In this study, we investigated whether MDA-MB-468 breast cancer cells induced to undergo EMT exhibit changes in mRNA levels of calcium channels, pumps and exchangers located on intracellular calcium storing organelles, including the Golgi, mitochondria and endoplasmic reticulum (ER).

Methods: Epidermal growth factor (EGF) was used to induce EMT in MDA-MB-468 breast cancer cells. Serum-deprived cells were treated with EGF (50 ng/mL) for 12 h and gene expression was assessed using quantitative RT-PCR.

Results and conclusions: These data reveal no significant alterations in mRNA levels of the Golgi calcium pump secretory pathway calcium ATPases (SPCA1 and SPCA2), or the mitochondrial calcium uniporter (MCU) or Na+/Ca2+ exchanger (NCLX). However, EGF-induced EMT was associated with significant alterations in mRNA levels of specific ER calcium channels and pumps, including (sarco)-endoplasmic reticulum calcium ATPases (SERCAs), and inositol 1,4,5-trisphosphate receptor (IP3R) and ryanodine receptor (RYR) calcium channel isoforms. The most prominent change in gene expression between the epithelial and mesenchymal-like states was RYR2, which was enriched 45-fold in EGF-treated MDA-MB-468 cells. These findings indicate that EGF-induced EMT in breast cancer cells may be associated with major alterations in ER calcium homeostasis.

No MeSH data available.


Related in: MedlinePlus