Limits...
The ATRA-induced differentiation of medulloblastoma cells is enhanced with LOX/COX inhibitors: an analysis of gene expression.

Chlapek P, Neradil J, Redova M, Zitterbart K, Sterba J, Veselska R - Cancer Cell Int. (2014)

Bottom Line: This effect was detected in both cell lines.This effect was also observed for the CDKN1A gene encoding the p21 protein, which is an important regulator of the cell cycle, and for the genes encoding proteins that are associated with proteasome activity.Furthermore, our results showed that D283 Med cells are significantly more sensitive to treatment with ATRA alone than Daoy cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Experimental Biology - Laboratory of Tumor Biology, School of Science, Masaryk University, Kotlarska 2, 611 37 Brno, Czech Republic.

ABSTRACT

Background: A detailed analysis of the expression of 440 cancer-related genes was performed after the combined treatment of medulloblastoma cells with all-trans retinoic acid (ATRA) and inhibitors of lipoxygenases (LOX) and cyclooxygenases (COX). The combinations of retinoids and celecoxib as a COX-2 inhibitor were reported to be effective in some regimens of metronomic therapy of relapsed solid tumors with poor prognosis. Our previous findings on neuroblastoma cells using expression profiling showed that LOX/COX inhibitors have the capability of enhancing the differentiating action of ATRA. Presented study focused on the continuation of our previous work to confirm the possibility of enhancing ATRA-induced cell differentiation in these cell lines via the application of LOX/COX inhibitors. This study provides more detailed information concerning the mechanisms of the enhancement of the ATRA-induced differentiation of medulloblastoma cells.

Methods: The Daoy and D283 Med medulloblastoma cell lines were chosen for this study. Caffeic acid (an inhibitor of 5-LOX) and celecoxib (an inhibitor on COX-2) were used in combined treatment with ATRA. The expression profiling was performed using Human Cancer Oligo GEArray membranes, and the most promising results were verified using RT-PCR.

Results: The expression profiling of the selected cancer-related genes clearly confirmed that the differentiating effects of ATRA should be enhanced via its combined administration with caffeic acid or celecoxib. This effect was detected in both cell lines. An increased expression of the genes that encoded the proteins participating in induced differentiation and cytoskeleton remodeling was detected in both cell lines in a concentration-dependent manner. This effect was also observed for the CDKN1A gene encoding the p21 protein, which is an important regulator of the cell cycle, and for the genes encoding proteins that are associated with proteasome activity. Furthermore, our results showed that D283 Med cells are significantly more sensitive to treatment with ATRA alone than Daoy cells.

Conclusions: The obtained results on medulloblastoma cell lines are in accordance with our previous findings on neuroblastoma cells and confirm our hypothesis concerning the common mechanism of the enhancement of ATRA-induced cell differentiation in various types of pediatric solid tumors.

No MeSH data available.


Related in: MedlinePlus

Changes in the expression of the GDF15 gene in Daoy (A) and D283 Med (B) cells analyzed using RT-PCR. The cells were treated with ATRA alone or in combination with CA (an inhibitor of 5-LOX) or CX (an inhibitor of COX-2); numbers indicate the concentrations of these compounds in μM. Respective numerical data from densitometry for three independent experiments are shown in the graphs: X-axis, treatment condition; Y-axis, relative gene expression. The data represent the means ± SD; they were normalized to the expression of the HSP90AB1 housekeeping gene and were related to the untreated control cells. Representative electrophoresis is presented for each cell line.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4066709&req=5

Figure 3: Changes in the expression of the GDF15 gene in Daoy (A) and D283 Med (B) cells analyzed using RT-PCR. The cells were treated with ATRA alone or in combination with CA (an inhibitor of 5-LOX) or CX (an inhibitor of COX-2); numbers indicate the concentrations of these compounds in μM. Respective numerical data from densitometry for three independent experiments are shown in the graphs: X-axis, treatment condition; Y-axis, relative gene expression. The data represent the means ± SD; they were normalized to the expression of the HSP90AB1 housekeeping gene and were related to the untreated control cells. Representative electrophoresis is presented for each cell line.

Mentions: The analysis of the detected changes in gene expressions showed a concentration-dependent increase in the expression of genes known to be involved in the process of retinoid-induced differentiation and especially of the genes generally associated with the regulation of the cell cycle, of genes involved in mitochondrial metabolism, etc. The observed changes in the expressions of two selected candidate genes - GDF15 (Figure 3) and CDKN1A (Figure 4) - were subsequently verified via RT-PCR.


The ATRA-induced differentiation of medulloblastoma cells is enhanced with LOX/COX inhibitors: an analysis of gene expression.

Chlapek P, Neradil J, Redova M, Zitterbart K, Sterba J, Veselska R - Cancer Cell Int. (2014)

Changes in the expression of the GDF15 gene in Daoy (A) and D283 Med (B) cells analyzed using RT-PCR. The cells were treated with ATRA alone or in combination with CA (an inhibitor of 5-LOX) or CX (an inhibitor of COX-2); numbers indicate the concentrations of these compounds in μM. Respective numerical data from densitometry for three independent experiments are shown in the graphs: X-axis, treatment condition; Y-axis, relative gene expression. The data represent the means ± SD; they were normalized to the expression of the HSP90AB1 housekeeping gene and were related to the untreated control cells. Representative electrophoresis is presented for each cell line.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4066709&req=5

Figure 3: Changes in the expression of the GDF15 gene in Daoy (A) and D283 Med (B) cells analyzed using RT-PCR. The cells were treated with ATRA alone or in combination with CA (an inhibitor of 5-LOX) or CX (an inhibitor of COX-2); numbers indicate the concentrations of these compounds in μM. Respective numerical data from densitometry for three independent experiments are shown in the graphs: X-axis, treatment condition; Y-axis, relative gene expression. The data represent the means ± SD; they were normalized to the expression of the HSP90AB1 housekeeping gene and were related to the untreated control cells. Representative electrophoresis is presented for each cell line.
Mentions: The analysis of the detected changes in gene expressions showed a concentration-dependent increase in the expression of genes known to be involved in the process of retinoid-induced differentiation and especially of the genes generally associated with the regulation of the cell cycle, of genes involved in mitochondrial metabolism, etc. The observed changes in the expressions of two selected candidate genes - GDF15 (Figure 3) and CDKN1A (Figure 4) - were subsequently verified via RT-PCR.

Bottom Line: This effect was detected in both cell lines.This effect was also observed for the CDKN1A gene encoding the p21 protein, which is an important regulator of the cell cycle, and for the genes encoding proteins that are associated with proteasome activity.Furthermore, our results showed that D283 Med cells are significantly more sensitive to treatment with ATRA alone than Daoy cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Experimental Biology - Laboratory of Tumor Biology, School of Science, Masaryk University, Kotlarska 2, 611 37 Brno, Czech Republic.

ABSTRACT

Background: A detailed analysis of the expression of 440 cancer-related genes was performed after the combined treatment of medulloblastoma cells with all-trans retinoic acid (ATRA) and inhibitors of lipoxygenases (LOX) and cyclooxygenases (COX). The combinations of retinoids and celecoxib as a COX-2 inhibitor were reported to be effective in some regimens of metronomic therapy of relapsed solid tumors with poor prognosis. Our previous findings on neuroblastoma cells using expression profiling showed that LOX/COX inhibitors have the capability of enhancing the differentiating action of ATRA. Presented study focused on the continuation of our previous work to confirm the possibility of enhancing ATRA-induced cell differentiation in these cell lines via the application of LOX/COX inhibitors. This study provides more detailed information concerning the mechanisms of the enhancement of the ATRA-induced differentiation of medulloblastoma cells.

Methods: The Daoy and D283 Med medulloblastoma cell lines were chosen for this study. Caffeic acid (an inhibitor of 5-LOX) and celecoxib (an inhibitor on COX-2) were used in combined treatment with ATRA. The expression profiling was performed using Human Cancer Oligo GEArray membranes, and the most promising results were verified using RT-PCR.

Results: The expression profiling of the selected cancer-related genes clearly confirmed that the differentiating effects of ATRA should be enhanced via its combined administration with caffeic acid or celecoxib. This effect was detected in both cell lines. An increased expression of the genes that encoded the proteins participating in induced differentiation and cytoskeleton remodeling was detected in both cell lines in a concentration-dependent manner. This effect was also observed for the CDKN1A gene encoding the p21 protein, which is an important regulator of the cell cycle, and for the genes encoding proteins that are associated with proteasome activity. Furthermore, our results showed that D283 Med cells are significantly more sensitive to treatment with ATRA alone than Daoy cells.

Conclusions: The obtained results on medulloblastoma cell lines are in accordance with our previous findings on neuroblastoma cells and confirm our hypothesis concerning the common mechanism of the enhancement of ATRA-induced cell differentiation in various types of pediatric solid tumors.

No MeSH data available.


Related in: MedlinePlus