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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 gene expression in explant cultures of patient-derived tissues after treatment with OXA or OXA + Curcumin.(a) Bar graph illustrating CXCL1 expression levels in 8 FFPE samples from explant cultures of CRC patients-derived liver metastases, that were treated with DMSO (NT), OXA or OXA plus Curcumin (CURC) for 24 h. (b) Scattergram reporting the expression levels of CXCL1 in NT, OXA or OXA plus Curcumin treated tissue explants. Horizontal lines represent mean values. (c) Table showing significative (S) or not significative (NS) changes in ki-67 (green) and cleaved caspase 3 (C3, blue) staining. Score 2 meant that ki-67 decreased and cleaved caspase increased significantly after the indicated treatment; score 1 meant that ki-67 decreased or cleaved caspase increased and score 0 meant that neither of them was changed. CXCL1 gene expression changes after NT, OXA or OXA + Curcumin treatment are also shown. (d) Scatter plot showing CXCL1 expression according to score values. Horizontal lines represent mean values. *p-value < 0.05 relative to NT (Wilcoxon test).
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f8: CXCL1 gene expression in explant cultures of patient-derived tissues after treatment with OXA or OXA + Curcumin.(a) Bar graph illustrating CXCL1 expression levels in 8 FFPE samples from explant cultures of CRC patients-derived liver metastases, that were treated with DMSO (NT), OXA or OXA plus Curcumin (CURC) for 24 h. (b) Scattergram reporting the expression levels of CXCL1 in NT, OXA or OXA plus Curcumin treated tissue explants. Horizontal lines represent mean values. (c) Table showing significative (S) or not significative (NS) changes in ki-67 (green) and cleaved caspase 3 (C3, blue) staining. Score 2 meant that ki-67 decreased and cleaved caspase increased significantly after the indicated treatment; score 1 meant that ki-67 decreased or cleaved caspase increased and score 0 meant that neither of them was changed. CXCL1 gene expression changes after NT, OXA or OXA + Curcumin treatment are also shown. (d) Scatter plot showing CXCL1 expression according to score values. Horizontal lines represent mean values. *p-value < 0.05 relative to NT (Wilcoxon test).

Mentions: In order to evaluate the potential of CXCL8 and CXCL1 as predictive biomarkers of OXA plus Curcumin treatment response, we analysed gene expression of both chemokines in a panel of 8 FFPE samples from explant cultures of CRC patients-derived liver metastases, that were treated with OXA or OXA plus Curcumin or vehicle (DMSO) for 24 h. Response to each treatment was assessed by means of decreasing proliferation and/or apoptosis activation as indicated by ki-67 and cleaved caspase-3 inmunohistochemistry staining, respectively34. The RNA concentrations obtained from these samples ranged (20–75 ng/μL) and under our qRT-PCR conditions we were unable to detect CXCL8 expression. According to our in vitro results, we observed an increase of CXCL1 expression in 6/8 explants treated with OXA that was attenuated by the addition of Curcumin in 5/6 explants (Fig. 8a,b). We then studied the expression patterns of CXCL1 according to “response” to treatment. To do this, we assigned each case a score according to changes in ki-67 and cleaved caspase 3 staining after treatments (compared to vehicle). Thus, a score of 2 meant that ki-67 decreased and cleaved caspase 3 increased significantly after treatment; score 1 meant that ki-67 decreased or cleaved caspase 3 increased and score 0 meant that neither of them was changed according to this criteria. Interestingly, other authors have used a similar approach before39. As shown in Fig. 8c, explants p93 and p141 were the only ones with a score = 2 when treated with Curcumin + OXA and interestingly, they had the highest CXCL1 basal levels as compared to explants with score 1 or 0. Although these results were obtained in a very few cases, they are in agreement with those obtained in vitro supporting the idea that those patients whose tumours present high CXCL1 levels (basally or after treatment with OXA-based chemotherapy) would be suitable candidates to be treated with a combination of Curcumin plus OXA (Fig. 8d). Whether this also applies to CXCL8, remains to be elucidated. Further clinical studies are on going in order to investigate these hypotheses.


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 gene expression in explant cultures of patient-derived tissues after treatment with OXA or OXA + Curcumin.(a) Bar graph illustrating CXCL1 expression levels in 8 FFPE samples from explant cultures of CRC patients-derived liver metastases, that were treated with DMSO (NT), OXA or OXA plus Curcumin (CURC) for 24 h. (b) Scattergram reporting the expression levels of CXCL1 in NT, OXA or OXA plus Curcumin treated tissue explants. Horizontal lines represent mean values. (c) Table showing significative (S) or not significative (NS) changes in ki-67 (green) and cleaved caspase 3 (C3, blue) staining. Score 2 meant that ki-67 decreased and cleaved caspase increased significantly after the indicated treatment; score 1 meant that ki-67 decreased or cleaved caspase increased and score 0 meant that neither of them was changed. CXCL1 gene expression changes after NT, OXA or OXA + Curcumin treatment are also shown. (d) Scatter plot showing CXCL1 expression according to score values. Horizontal lines represent mean values. *p-value < 0.05 relative to NT (Wilcoxon test).
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Related In: Results  -  Collection

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f8: CXCL1 gene expression in explant cultures of patient-derived tissues after treatment with OXA or OXA + Curcumin.(a) Bar graph illustrating CXCL1 expression levels in 8 FFPE samples from explant cultures of CRC patients-derived liver metastases, that were treated with DMSO (NT), OXA or OXA plus Curcumin (CURC) for 24 h. (b) Scattergram reporting the expression levels of CXCL1 in NT, OXA or OXA plus Curcumin treated tissue explants. Horizontal lines represent mean values. (c) Table showing significative (S) or not significative (NS) changes in ki-67 (green) and cleaved caspase 3 (C3, blue) staining. Score 2 meant that ki-67 decreased and cleaved caspase increased significantly after the indicated treatment; score 1 meant that ki-67 decreased or cleaved caspase increased and score 0 meant that neither of them was changed. CXCL1 gene expression changes after NT, OXA or OXA + Curcumin treatment are also shown. (d) Scatter plot showing CXCL1 expression according to score values. Horizontal lines represent mean values. *p-value < 0.05 relative to NT (Wilcoxon test).
Mentions: In order to evaluate the potential of CXCL8 and CXCL1 as predictive biomarkers of OXA plus Curcumin treatment response, we analysed gene expression of both chemokines in a panel of 8 FFPE samples from explant cultures of CRC patients-derived liver metastases, that were treated with OXA or OXA plus Curcumin or vehicle (DMSO) for 24 h. Response to each treatment was assessed by means of decreasing proliferation and/or apoptosis activation as indicated by ki-67 and cleaved caspase-3 inmunohistochemistry staining, respectively34. The RNA concentrations obtained from these samples ranged (20–75 ng/μL) and under our qRT-PCR conditions we were unable to detect CXCL8 expression. According to our in vitro results, we observed an increase of CXCL1 expression in 6/8 explants treated with OXA that was attenuated by the addition of Curcumin in 5/6 explants (Fig. 8a,b). We then studied the expression patterns of CXCL1 according to “response” to treatment. To do this, we assigned each case a score according to changes in ki-67 and cleaved caspase 3 staining after treatments (compared to vehicle). Thus, a score of 2 meant that ki-67 decreased and cleaved caspase 3 increased significantly after treatment; score 1 meant that ki-67 decreased or cleaved caspase 3 increased and score 0 meant that neither of them was changed according to this criteria. Interestingly, other authors have used a similar approach before39. As shown in Fig. 8c, explants p93 and p141 were the only ones with a score = 2 when treated with Curcumin + OXA and interestingly, they had the highest CXCL1 basal levels as compared to explants with score 1 or 0. Although these results were obtained in a very few cases, they are in agreement with those obtained in vitro supporting the idea that those patients whose tumours present high CXCL1 levels (basally or after treatment with OXA-based chemotherapy) would be suitable candidates to be treated with a combination of Curcumin plus OXA (Fig. 8d). Whether this also applies to CXCL8, remains to be elucidated. Further clinical studies are on going in order to investigate these hypotheses.

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