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Celecoxib Up Regulates the Expression of Drug Efflux Transporter ABCG2 in Breast Cancer Cell Lines.

Kalalinia F, Elahian F, Mosaffa F, Behravan J - Iran J Pharm Res (2014)

Bottom Line: To clarify whether ABCG2 inhibition is involved in the sensitizing effect of celecoxib, we investigated whether the expression of ABCG2 in breast cancer cell lines, could be modulated by celecoxib.The COX-2 inhibitor celecoxib up-regulated the expression of ABCG2 mRNA in MCF-7 and MCF7-MX cells, which was accompanied by increased ABCG2 protein expression.Our findings provide evidence that celecoxib up-regulates ABCG2 expression in human breast cancer cells and proposed that ABCG2 is not involved in chemosensitizing effects of celecoxib.

View Article: PubMed Central - PubMed

Affiliation: Biotechnology Research Center, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, IRAN. ; Department of Pharmaceutical Biotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, IRAN.

ABSTRACT
Elevated expression of the drug efflux transporter ABCG2 seems to correlate with multidrug resistance of cancer cells. Specific COX-2 inhibitor celecoxib has been shown to enhance the sensitivity of cancer cells to anticancer drugs. To clarify whether ABCG2 inhibition is involved in the sensitizing effect of celecoxib, we investigated whether the expression of ABCG2 in breast cancer cell lines, could be modulated by celecoxib. The expression of the multidrug resistant gene (ABCG2) at mRNA and protein level was detected by real-time quantitative reverse transcription-polymerase chain reaction and flow cytometry analysis, respectively. Among three human breast cancer cell lines ABCG2 and COX-2 were highly expressed in MCF7-MX and MDA-MB-231 cells, respectively. The COX-2 inhibitor celecoxib up-regulated the expression of ABCG2 mRNA in MCF-7 and MCF7-MX cells, which was accompanied by increased ABCG2 protein expression. While celecoxib was able to block the 12-O-tetradecanoylphorbol-13-acetate (TPA)-mediated increase in COX-2 expression in MDA-MB-231 cells, it increased the expression of ABCG2 up to 4.27 times to the control level at mRNA level and with less intensity at protein level. Our findings provide evidence that celecoxib up-regulates ABCG2 expression in human breast cancer cells and proposed that ABCG2 is not involved in chemosensitizing effects of celecoxib.

No MeSH data available.


Related in: MedlinePlus

COX-2 mRNA expression in MDA-MB-231 cell line under treatment with and without celecoxib. Line chart shows the results of cells that were treated with TPA 10 nM lonely. Bar chart indicated the results of cells that were treated with TPA 10 nM in combination with celecoxib 0-40 µM for 4-24 h. Real-time RT-PCR analysis was performed on total RNA extracted from control and treated cells. Values were normalized to the β-actin content of samples. The results were expressed as the target/reference ratio of the treated samples divided by the target/reference ratio of the untreated control sample and expressed as mean ± SD (n = 3); *, p < 0.05; **, p < 0.01; ***, p < 0.001
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Figure 2: COX-2 mRNA expression in MDA-MB-231 cell line under treatment with and without celecoxib. Line chart shows the results of cells that were treated with TPA 10 nM lonely. Bar chart indicated the results of cells that were treated with TPA 10 nM in combination with celecoxib 0-40 µM for 4-24 h. Real-time RT-PCR analysis was performed on total RNA extracted from control and treated cells. Values were normalized to the β-actin content of samples. The results were expressed as the target/reference ratio of the treated samples divided by the target/reference ratio of the untreated control sample and expressed as mean ± SD (n = 3); *, p < 0.05; **, p < 0.01; ***, p < 0.001

Mentions: To evaluate the expression of COX-2 mRNA under celecoxib treatment, three cell lines were treated with non-toxic concentration of TPA (10 nM) and celecoxib (0-40 µM) for 4-24 h (15). Expression of COX-2 was studied using Real-time RT-PCR. We have previously reported that TPA was a potent stimulator of COX-2 expression in breast cancer cell lines; as it stimulates COX-2 expression up to 4.5 fold vs. control by 4 h in MDA-MB-231 cells (15). Figure 2 indicates that celecoxib reverses the effects of TPA on COX-2 expression to the control level in MDA-MB-231 by 4 h. Celecoxib significantly inhibited COX-2 expression by 12 and 24 h in MCF-7 and by 24 h in MCF7-MX (data not shown). To verify whether the inhibitory effects of celecoxib on COX-2 expression could be observed also in protein level, cells were treated for 4 and 24 h without or with TPA 10 nM, celecoxib 40 µM, or both. Then, the cells were incubated with the specific COX-2 antibody. As shown in Figure 3, MDA-MB-231 cells treated with TPA displayed an increased COX-2 protein expression, which was significantly reduced by co-treatment with celecoxib.


Celecoxib Up Regulates the Expression of Drug Efflux Transporter ABCG2 in Breast Cancer Cell Lines.

Kalalinia F, Elahian F, Mosaffa F, Behravan J - Iran J Pharm Res (2014)

COX-2 mRNA expression in MDA-MB-231 cell line under treatment with and without celecoxib. Line chart shows the results of cells that were treated with TPA 10 nM lonely. Bar chart indicated the results of cells that were treated with TPA 10 nM in combination with celecoxib 0-40 µM for 4-24 h. Real-time RT-PCR analysis was performed on total RNA extracted from control and treated cells. Values were normalized to the β-actin content of samples. The results were expressed as the target/reference ratio of the treated samples divided by the target/reference ratio of the untreated control sample and expressed as mean ± SD (n = 3); *, p < 0.05; **, p < 0.01; ***, p < 0.001
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Related In: Results  -  Collection

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Figure 2: COX-2 mRNA expression in MDA-MB-231 cell line under treatment with and without celecoxib. Line chart shows the results of cells that were treated with TPA 10 nM lonely. Bar chart indicated the results of cells that were treated with TPA 10 nM in combination with celecoxib 0-40 µM for 4-24 h. Real-time RT-PCR analysis was performed on total RNA extracted from control and treated cells. Values were normalized to the β-actin content of samples. The results were expressed as the target/reference ratio of the treated samples divided by the target/reference ratio of the untreated control sample and expressed as mean ± SD (n = 3); *, p < 0.05; **, p < 0.01; ***, p < 0.001
Mentions: To evaluate the expression of COX-2 mRNA under celecoxib treatment, three cell lines were treated with non-toxic concentration of TPA (10 nM) and celecoxib (0-40 µM) for 4-24 h (15). Expression of COX-2 was studied using Real-time RT-PCR. We have previously reported that TPA was a potent stimulator of COX-2 expression in breast cancer cell lines; as it stimulates COX-2 expression up to 4.5 fold vs. control by 4 h in MDA-MB-231 cells (15). Figure 2 indicates that celecoxib reverses the effects of TPA on COX-2 expression to the control level in MDA-MB-231 by 4 h. Celecoxib significantly inhibited COX-2 expression by 12 and 24 h in MCF-7 and by 24 h in MCF7-MX (data not shown). To verify whether the inhibitory effects of celecoxib on COX-2 expression could be observed also in protein level, cells were treated for 4 and 24 h without or with TPA 10 nM, celecoxib 40 µM, or both. Then, the cells were incubated with the specific COX-2 antibody. As shown in Figure 3, MDA-MB-231 cells treated with TPA displayed an increased COX-2 protein expression, which was significantly reduced by co-treatment with celecoxib.

Bottom Line: To clarify whether ABCG2 inhibition is involved in the sensitizing effect of celecoxib, we investigated whether the expression of ABCG2 in breast cancer cell lines, could be modulated by celecoxib.The COX-2 inhibitor celecoxib up-regulated the expression of ABCG2 mRNA in MCF-7 and MCF7-MX cells, which was accompanied by increased ABCG2 protein expression.Our findings provide evidence that celecoxib up-regulates ABCG2 expression in human breast cancer cells and proposed that ABCG2 is not involved in chemosensitizing effects of celecoxib.

View Article: PubMed Central - PubMed

Affiliation: Biotechnology Research Center, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, IRAN. ; Department of Pharmaceutical Biotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, IRAN.

ABSTRACT
Elevated expression of the drug efflux transporter ABCG2 seems to correlate with multidrug resistance of cancer cells. Specific COX-2 inhibitor celecoxib has been shown to enhance the sensitivity of cancer cells to anticancer drugs. To clarify whether ABCG2 inhibition is involved in the sensitizing effect of celecoxib, we investigated whether the expression of ABCG2 in breast cancer cell lines, could be modulated by celecoxib. The expression of the multidrug resistant gene (ABCG2) at mRNA and protein level was detected by real-time quantitative reverse transcription-polymerase chain reaction and flow cytometry analysis, respectively. Among three human breast cancer cell lines ABCG2 and COX-2 were highly expressed in MCF7-MX and MDA-MB-231 cells, respectively. The COX-2 inhibitor celecoxib up-regulated the expression of ABCG2 mRNA in MCF-7 and MCF7-MX cells, which was accompanied by increased ABCG2 protein expression. While celecoxib was able to block the 12-O-tetradecanoylphorbol-13-acetate (TPA)-mediated increase in COX-2 expression in MDA-MB-231 cells, it increased the expression of ABCG2 up to 4.27 times to the control level at mRNA level and with less intensity at protein level. Our findings provide evidence that celecoxib up-regulates ABCG2 expression in human breast cancer cells and proposed that ABCG2 is not involved in chemosensitizing effects of celecoxib.

No MeSH data available.


Related in: MedlinePlus