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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

CXCL1, CXCL8 and CXCL2 gene expression and CXCL1 and CXCL8 secreted protein levels after treatment with OXA, Curcumin or both.(a) Bar graphs illustrating relative gene expression levels (mean ± SEM) of the indicated chemokines in non-treated (NT) and after OXA or OXA plus Curcumin (CURC) treatment at IC50 doses in HT29 and HTOXAR3 cells. Gene expression levels of β-actin were used as endogenous control. *p-value < 0.05 relative to NT, #p-value < 0.05; ##p-value < 0.01, relative to HT29. ** p-value < 0.05 relative to OXA. (b) Changes in chemokines’ gene expression after treatment with Curcumin at indicated doses. (c) Graph bars showing levels of secreted chemokines (mean ± SEM) measured by enzyme-linked immunosorbent assay (ELISA) in the supernatants of treated cells as indicated. *p-value < 0.05; **p-value < 0.01; ***p-value < 0.001 relative to NT. ##p-value < 0.01, ###p-value < 0,001 relative to OXA. Results were obtained from at least 3 independent experiments.
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f6: CXCL1, CXCL8 and CXCL2 gene expression and CXCL1 and CXCL8 secreted protein levels after treatment with OXA, Curcumin or both.(a) Bar graphs illustrating relative gene expression levels (mean ± SEM) of the indicated chemokines in non-treated (NT) and after OXA or OXA plus Curcumin (CURC) treatment at IC50 doses in HT29 and HTOXAR3 cells. Gene expression levels of β-actin were used as endogenous control. *p-value < 0.05 relative to NT, #p-value < 0.05; ##p-value < 0.01, relative to HT29. ** p-value < 0.05 relative to OXA. (b) Changes in chemokines’ gene expression after treatment with Curcumin at indicated doses. (c) Graph bars showing levels of secreted chemokines (mean ± SEM) measured by enzyme-linked immunosorbent assay (ELISA) in the supernatants of treated cells as indicated. *p-value < 0.05; **p-value < 0.01; ***p-value < 0.001 relative to NT. ##p-value < 0.01, ###p-value < 0,001 relative to OXA. Results were obtained from at least 3 independent experiments.

Mentions: Among the top 10 genes fitting these criteria (Fig. 5) we observed that three pro-inflammatory CXC-chemokines, CXCL8 (Interleukin-8), CXCL1 (Gro-α) and CXCL2 (Gro-β) that are primarily regulated by NF-κB35 were significantly up-regulated at basal level, and were more efficiently down-regulated after treatment with OXA plus Curcumin in the resistant cell line. Accordingly, functional enrichment analysis revealed cytokine signalling as one of the two molecular functions significantly overrepresented in this condition (Supplementary Figure S5). These results were validated through qRT-PCR experiments, confirming a basal overexpression of CXCL8 (~10-fold), CXCL1 (~6-fold) and CXCL2 (~2.5-fold) in HTOXAR3 cell line as compared with HT29 (Fig. 6a). The three chemokines were up-regulated after OXA treatment only in HT29 cells (CXCL8 3-fold, CXCL1 6-fold and CXCL2 2-fold; Fig. 6a). In the case of CXCL2, we observed a down-regulation in expression after OXA treatment in HTOXAR3 cells. Interestingly, the OXA-induced expression of CXCL8 and CXCL1 was significantly attenuated by the addition of Curcumin in the resistant cell line but not in the sensitive one (Fig. 6a). Down-regulation of CXCL2 with regards to basal conditions was also observed in the HTOXAR3 cell line after concomitant treatment (Fig. 6a). In view of these results, we investigated the effect of a 24 h-treatment with Curcumin as a single agent at different doses, on the expression of CXCL8 and CXCL1 observing a significant reduction in a dose-dependent manner only in the resistant cell line while in HT29 cells, this reduction was only observed at very high doses (Fig. 6b). ELISA-based analysis of HT29 and HTOXAR3 conditioned media revealed similar results at protein-secreted levels (Fig. 6c).


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)

CXCL1, CXCL8 and CXCL2 gene expression and CXCL1 and CXCL8 secreted protein levels after treatment with OXA, Curcumin or both.(a) Bar graphs illustrating relative gene expression levels (mean ± SEM) of the indicated chemokines in non-treated (NT) and after OXA or OXA plus Curcumin (CURC) treatment at IC50 doses in HT29 and HTOXAR3 cells. Gene expression levels of β-actin were used as endogenous control. *p-value < 0.05 relative to NT, #p-value < 0.05; ##p-value < 0.01, relative to HT29. ** p-value < 0.05 relative to OXA. (b) Changes in chemokines’ gene expression after treatment with Curcumin at indicated doses. (c) Graph bars showing levels of secreted chemokines (mean ± SEM) measured by enzyme-linked immunosorbent assay (ELISA) in the supernatants of treated cells as indicated. *p-value < 0.05; **p-value < 0.01; ***p-value < 0.001 relative to NT. ##p-value < 0.01, ###p-value < 0,001 relative to OXA. Results were obtained from at least 3 independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4835769&req=5

f6: CXCL1, CXCL8 and CXCL2 gene expression and CXCL1 and CXCL8 secreted protein levels after treatment with OXA, Curcumin or both.(a) Bar graphs illustrating relative gene expression levels (mean ± SEM) of the indicated chemokines in non-treated (NT) and after OXA or OXA plus Curcumin (CURC) treatment at IC50 doses in HT29 and HTOXAR3 cells. Gene expression levels of β-actin were used as endogenous control. *p-value < 0.05 relative to NT, #p-value < 0.05; ##p-value < 0.01, relative to HT29. ** p-value < 0.05 relative to OXA. (b) Changes in chemokines’ gene expression after treatment with Curcumin at indicated doses. (c) Graph bars showing levels of secreted chemokines (mean ± SEM) measured by enzyme-linked immunosorbent assay (ELISA) in the supernatants of treated cells as indicated. *p-value < 0.05; **p-value < 0.01; ***p-value < 0.001 relative to NT. ##p-value < 0.01, ###p-value < 0,001 relative to OXA. Results were obtained from at least 3 independent experiments.
Mentions: Among the top 10 genes fitting these criteria (Fig. 5) we observed that three pro-inflammatory CXC-chemokines, CXCL8 (Interleukin-8), CXCL1 (Gro-α) and CXCL2 (Gro-β) that are primarily regulated by NF-κB35 were significantly up-regulated at basal level, and were more efficiently down-regulated after treatment with OXA plus Curcumin in the resistant cell line. Accordingly, functional enrichment analysis revealed cytokine signalling as one of the two molecular functions significantly overrepresented in this condition (Supplementary Figure S5). These results were validated through qRT-PCR experiments, confirming a basal overexpression of CXCL8 (~10-fold), CXCL1 (~6-fold) and CXCL2 (~2.5-fold) in HTOXAR3 cell line as compared with HT29 (Fig. 6a). The three chemokines were up-regulated after OXA treatment only in HT29 cells (CXCL8 3-fold, CXCL1 6-fold and CXCL2 2-fold; Fig. 6a). In the case of CXCL2, we observed a down-regulation in expression after OXA treatment in HTOXAR3 cells. Interestingly, the OXA-induced expression of CXCL8 and CXCL1 was significantly attenuated by the addition of Curcumin in the resistant cell line but not in the sensitive one (Fig. 6a). Down-regulation of CXCL2 with regards to basal conditions was also observed in the HTOXAR3 cell line after concomitant treatment (Fig. 6a). In view of these results, we investigated the effect of a 24 h-treatment with Curcumin as a single agent at different doses, on the expression of CXCL8 and CXCL1 observing a significant reduction in a dose-dependent manner only in the resistant cell line while in HT29 cells, this reduction was only observed at very high doses (Fig. 6b). ELISA-based analysis of HT29 and HTOXAR3 conditioned media revealed similar results at protein-secreted levels (Fig. 6c).

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