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Curcumin mediates oxaliplatin-acquired resistance reversion in colorectal cancer cell lines through modulation of CXC-Chemokine/NF-κB signalling pathway.

Ruiz de Porras V, Bystrup S, Martínez-Cardús A, Pluvinet R, Sumoy L, Howells L, James MI, Iwuji C, Manzano JL, Layos L, Bugés C, Abad A, Martínez-Balibrea E - Sci Rep (2016)

Bottom Line: Transcriptomic profiling revealed the up-regulation of three NF-κB-regulated CXC-chemokines, CXCL8, CXCL1 and CXCL2, in the resistant cells that were more efficiently down-regulated after OXA + Curcumin treatment as compared to the sensitive cells.High expression of CXCL1 in FFPE samples from explant cultures of CRC patients-derived liver metastases was associated with response to OXA + Curcumin.In conclusion, we suggest that combination of OXA + Curcumin could be an effective treatment, for which CXCL1 could be used as a predictive marker, in CRC patients.

View Article: PubMed Central - PubMed

Affiliation: Health Sciences Research Institute of the Germans Trias i Pujol Foundation (IGTP), Can Ruti Campus, Ctra. Can Ruti- Camí de les escoles s/n, 08916, Badalona, Spain.

ABSTRACT
Resistance to oxaliplatin (OXA) is a complex process affecting the outcomes of metastatic colorectal cancer (CRC) patients treated with this drug. De-regulation of the NF-κB signalling pathway has been proposed as an important mechanism involved in this phenomenon. Here, we show that NF-κB was hyperactivated in in vitro models of OXA-acquired resistance but was attenuated by the addition of Curcumin, a non-toxic NF-κB inhibitor. The concomitant combination of Curcumin + OXA was more effective and synergistic in cell lines with acquired resistance to OXA, leading to the reversion of their resistant phenotype, through the inhibition of the NF-κB signalling cascade. Transcriptomic profiling revealed the up-regulation of three NF-κB-regulated CXC-chemokines, CXCL8, CXCL1 and CXCL2, in the resistant cells that were more efficiently down-regulated after OXA + Curcumin treatment as compared to the sensitive cells. Moreover, CXCL8 and CXCL1 gene silencing made resistant cells more sensitive to OXA through the inhibition of the Akt/NF-κB pathway. High expression of CXCL1 in FFPE samples from explant cultures of CRC patients-derived liver metastases was associated with response to OXA + Curcumin. In conclusion, we suggest that combination of OXA + Curcumin could be an effective treatment, for which CXCL1 could be used as a predictive marker, in CRC patients.

No MeSH data available.


Related in: MedlinePlus

Effect of Curcumin on constitutive and OXA-induced NF-κB activation.(a) Western blot images of phosphorylated p65 (p-p65 Ser536), total p65, Survivin and Bcl-2 in HT29 and HTOXAR3 cells after a 24-hour treatment with 10 and 20 μM Curcumin (CURC). (b) Representative western blot images showing protein expression changes as indicated, in HT29 and HTOXAR3 cells after 24 h treatment with OXA or concomitant OXA plus Curcumin, for 0 to 120 minutes or 24 hours (c,d) Western blots showing p65 protein expression in cytoplasmic and nuclear extracts of HT29 cells after treatment with OXA (10 μM) or OXA + Curcumin (10 μM each) for short-time exposure (0 to 30 min) or 24 h (e). TNFα was used as positive control at 50 ng/ml. Msh2 and α-tubulin were used as nuclear- and cytoplasmic-specific protein controls, respectively. Images are representative of at least 3 independent experiments. NT: Non-treated cells.
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f4: Effect of Curcumin on constitutive and OXA-induced NF-κB activation.(a) Western blot images of phosphorylated p65 (p-p65 Ser536), total p65, Survivin and Bcl-2 in HT29 and HTOXAR3 cells after a 24-hour treatment with 10 and 20 μM Curcumin (CURC). (b) Representative western blot images showing protein expression changes as indicated, in HT29 and HTOXAR3 cells after 24 h treatment with OXA or concomitant OXA plus Curcumin, for 0 to 120 minutes or 24 hours (c,d) Western blots showing p65 protein expression in cytoplasmic and nuclear extracts of HT29 cells after treatment with OXA (10 μM) or OXA + Curcumin (10 μM each) for short-time exposure (0 to 30 min) or 24 h (e). TNFα was used as positive control at 50 ng/ml. Msh2 and α-tubulin were used as nuclear- and cytoplasmic-specific protein controls, respectively. Images are representative of at least 3 independent experiments. NT: Non-treated cells.

Mentions: In view of our results, we wanted to study the effect of the addition of Curcumin on OXA treatment at the molecular level. We first investigated the effect of Curcumin as a single agent on NF-κB inhibition. HT29 and HTOXAR3 cells were treated with Curcumin at 10 and 20 μM for 24 h. As expected, Curcumin decreased p65 phosphorylation and Survivin expression in a dose-dependent manner in both cell lines (Fig. 4a). Next, both cell lines were treated with OXA and OXA plus Curcumin at IC50 doses for short exposure times (0 to 120 min) and for 24 h. Results in Fig. 4b show that, in HT29/HTOXAR3 cells, the addition of Curcumin significantly decreased the OXA-induced phosphorylation of p65 (p < 0.001/p = 0.04) and IκBα (p < 0.05/p = 0.02) as well as the expression of Bcl-2 (p < 0.03/p < 0.01), Survivin (p < 0.001/p = 0.02) and Cyclin D1 (p < 0.001/p = 0.04) after short treatment times. Similar results were observed after a 24 h-treatment, although they did not reach statistical significance (Fig. 4c). We also studied how the addition of Curcumin affected the OXA-induced p65 nuclear translocation. As shown in Fig. 4d,e, levels of nuclear p65 were decreased in all cases when HT29 cells were treated with OXA plus Curcumin as compared with OXA alone.


Curcumin mediates oxaliplatin-acquired resistance reversion in colorectal cancer cell lines through modulation of CXC-Chemokine/NF-κB signalling pathway.

Ruiz de Porras V, Bystrup S, Martínez-Cardús A, Pluvinet R, Sumoy L, Howells L, James MI, Iwuji C, Manzano JL, Layos L, Bugés C, Abad A, Martínez-Balibrea E - Sci Rep (2016)

Effect of Curcumin on constitutive and OXA-induced NF-κB activation.(a) Western blot images of phosphorylated p65 (p-p65 Ser536), total p65, Survivin and Bcl-2 in HT29 and HTOXAR3 cells after a 24-hour treatment with 10 and 20 μM Curcumin (CURC). (b) Representative western blot images showing protein expression changes as indicated, in HT29 and HTOXAR3 cells after 24 h treatment with OXA or concomitant OXA plus Curcumin, for 0 to 120 minutes or 24 hours (c,d) Western blots showing p65 protein expression in cytoplasmic and nuclear extracts of HT29 cells after treatment with OXA (10 μM) or OXA + Curcumin (10 μM each) for short-time exposure (0 to 30 min) or 24 h (e). TNFα was used as positive control at 50 ng/ml. Msh2 and α-tubulin were used as nuclear- and cytoplasmic-specific protein controls, respectively. Images are representative of at least 3 independent experiments. NT: Non-treated cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4835769&req=5

f4: Effect of Curcumin on constitutive and OXA-induced NF-κB activation.(a) Western blot images of phosphorylated p65 (p-p65 Ser536), total p65, Survivin and Bcl-2 in HT29 and HTOXAR3 cells after a 24-hour treatment with 10 and 20 μM Curcumin (CURC). (b) Representative western blot images showing protein expression changes as indicated, in HT29 and HTOXAR3 cells after 24 h treatment with OXA or concomitant OXA plus Curcumin, for 0 to 120 minutes or 24 hours (c,d) Western blots showing p65 protein expression in cytoplasmic and nuclear extracts of HT29 cells after treatment with OXA (10 μM) or OXA + Curcumin (10 μM each) for short-time exposure (0 to 30 min) or 24 h (e). TNFα was used as positive control at 50 ng/ml. Msh2 and α-tubulin were used as nuclear- and cytoplasmic-specific protein controls, respectively. Images are representative of at least 3 independent experiments. NT: Non-treated cells.
Mentions: In view of our results, we wanted to study the effect of the addition of Curcumin on OXA treatment at the molecular level. We first investigated the effect of Curcumin as a single agent on NF-κB inhibition. HT29 and HTOXAR3 cells were treated with Curcumin at 10 and 20 μM for 24 h. As expected, Curcumin decreased p65 phosphorylation and Survivin expression in a dose-dependent manner in both cell lines (Fig. 4a). Next, both cell lines were treated with OXA and OXA plus Curcumin at IC50 doses for short exposure times (0 to 120 min) and for 24 h. Results in Fig. 4b show that, in HT29/HTOXAR3 cells, the addition of Curcumin significantly decreased the OXA-induced phosphorylation of p65 (p < 0.001/p = 0.04) and IκBα (p < 0.05/p = 0.02) as well as the expression of Bcl-2 (p < 0.03/p < 0.01), Survivin (p < 0.001/p = 0.02) and Cyclin D1 (p < 0.001/p = 0.04) after short treatment times. Similar results were observed after a 24 h-treatment, although they did not reach statistical significance (Fig. 4c). We also studied how the addition of Curcumin affected the OXA-induced p65 nuclear translocation. As shown in Fig. 4d,e, levels of nuclear p65 were decreased in all cases when HT29 cells were treated with OXA plus Curcumin as compared with OXA alone.

Bottom Line: Transcriptomic profiling revealed the up-regulation of three NF-κB-regulated CXC-chemokines, CXCL8, CXCL1 and CXCL2, in the resistant cells that were more efficiently down-regulated after OXA + Curcumin treatment as compared to the sensitive cells.High expression of CXCL1 in FFPE samples from explant cultures of CRC patients-derived liver metastases was associated with response to OXA + Curcumin.In conclusion, we suggest that combination of OXA + Curcumin could be an effective treatment, for which CXCL1 could be used as a predictive marker, in CRC patients.

View Article: PubMed Central - PubMed

Affiliation: Health Sciences Research Institute of the Germans Trias i Pujol Foundation (IGTP), Can Ruti Campus, Ctra. Can Ruti- Camí de les escoles s/n, 08916, Badalona, Spain.

ABSTRACT
Resistance to oxaliplatin (OXA) is a complex process affecting the outcomes of metastatic colorectal cancer (CRC) patients treated with this drug. De-regulation of the NF-κB signalling pathway has been proposed as an important mechanism involved in this phenomenon. Here, we show that NF-κB was hyperactivated in in vitro models of OXA-acquired resistance but was attenuated by the addition of Curcumin, a non-toxic NF-κB inhibitor. The concomitant combination of Curcumin + OXA was more effective and synergistic in cell lines with acquired resistance to OXA, leading to the reversion of their resistant phenotype, through the inhibition of the NF-κB signalling cascade. Transcriptomic profiling revealed the up-regulation of three NF-κB-regulated CXC-chemokines, CXCL8, CXCL1 and CXCL2, in the resistant cells that were more efficiently down-regulated after OXA + Curcumin treatment as compared to the sensitive cells. Moreover, CXCL8 and CXCL1 gene silencing made resistant cells more sensitive to OXA through the inhibition of the Akt/NF-κB pathway. High expression of CXCL1 in FFPE samples from explant cultures of CRC patients-derived liver metastases was associated with response to OXA + Curcumin. In conclusion, we suggest that combination of OXA + Curcumin could be an effective treatment, for which CXCL1 could be used as a predictive marker, in CRC patients.

No MeSH data available.


Related in: MedlinePlus