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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 CXCL8 and CXCL1 siRNA-mediated gene silencing on OXA cytotoxicity and NF-kB signalling pathway activation in HTOXR3 cells.(a) Dose-response curves for HTOXAR3 cell line, after CXCL8 and (b) CXCL1 gene silencing, treated with 0-100 μM OXA for 24 hours (mean ± SEM). Insets show siRNA-mediated gene silencing efficiency at 48 h post-transfection. IC50 values are indicated as mean (95% CI). (c) Representative western blot images showing changes in phosphorylated IκBα (p-IκBα Ser32/36), phosphorylated Akt (p-Akt Ser473), Survivin, Bcl-2 and Cyclin D1 protein expression in HTOXAR3 cells under control (siNTC) and CXCL8 (siCXCL8) or (d) CXCL1 (siCXCL1) gene silencing. All results were obtained from at least 3 independent experiments.
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f7: Effect of CXCL8 and CXCL1 siRNA-mediated gene silencing on OXA cytotoxicity and NF-kB signalling pathway activation in HTOXR3 cells.(a) Dose-response curves for HTOXAR3 cell line, after CXCL8 and (b) CXCL1 gene silencing, treated with 0-100 μM OXA for 24 hours (mean ± SEM). Insets show siRNA-mediated gene silencing efficiency at 48 h post-transfection. IC50 values are indicated as mean (95% CI). (c) Representative western blot images showing changes in phosphorylated IκBα (p-IκBα Ser32/36), phosphorylated Akt (p-Akt Ser473), Survivin, Bcl-2 and Cyclin D1 protein expression in HTOXAR3 cells under control (siNTC) and CXCL8 (siCXCL8) or (d) CXCL1 (siCXCL1) gene silencing. All results were obtained from at least 3 independent experiments.

Mentions: To confirm the specific role of CXCL8 and CXCL1 on OXA resistance acquisition in HTOXAR3 cells, we transiently silenced their gene expression by using siRNA oligonucleotides. We assessed the cytotoxicity of OXA by MTT assay in control cells (siNTC) and CXCL8 or CXCL1 knockdown cells (siCXCL8 or siCXCL1, respectively). Forty-eight hours after gene silencing, cells were seeded in 96-well plates and treated for 24 h with doses of OXA ranging 0–100 μM. CXCL8 and CXCL1 knockdown efficiency was ~90% and ~60% at mRNA level, respectively (insets on Fig. 7a,b). Under these conditions, we found that CXCL8 and CXCL1 gene silencing in HTOXAR3 led to a 35% and a 20% decrease in OXA IC50 value as compared to siNTC cells, respectively (Fig. 7a,b).


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 CXCL8 and CXCL1 siRNA-mediated gene silencing on OXA cytotoxicity and NF-kB signalling pathway activation in HTOXR3 cells.(a) Dose-response curves for HTOXAR3 cell line, after CXCL8 and (b) CXCL1 gene silencing, treated with 0-100 μM OXA for 24 hours (mean ± SEM). Insets show siRNA-mediated gene silencing efficiency at 48 h post-transfection. IC50 values are indicated as mean (95% CI). (c) Representative western blot images showing changes in phosphorylated IκBα (p-IκBα Ser32/36), phosphorylated Akt (p-Akt Ser473), Survivin, Bcl-2 and Cyclin D1 protein expression in HTOXAR3 cells under control (siNTC) and CXCL8 (siCXCL8) or (d) CXCL1 (siCXCL1) gene silencing. All results were obtained from at least 3 independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f7: Effect of CXCL8 and CXCL1 siRNA-mediated gene silencing on OXA cytotoxicity and NF-kB signalling pathway activation in HTOXR3 cells.(a) Dose-response curves for HTOXAR3 cell line, after CXCL8 and (b) CXCL1 gene silencing, treated with 0-100 μM OXA for 24 hours (mean ± SEM). Insets show siRNA-mediated gene silencing efficiency at 48 h post-transfection. IC50 values are indicated as mean (95% CI). (c) Representative western blot images showing changes in phosphorylated IκBα (p-IκBα Ser32/36), phosphorylated Akt (p-Akt Ser473), Survivin, Bcl-2 and Cyclin D1 protein expression in HTOXAR3 cells under control (siNTC) and CXCL8 (siCXCL8) or (d) CXCL1 (siCXCL1) gene silencing. All results were obtained from at least 3 independent experiments.
Mentions: To confirm the specific role of CXCL8 and CXCL1 on OXA resistance acquisition in HTOXAR3 cells, we transiently silenced their gene expression by using siRNA oligonucleotides. We assessed the cytotoxicity of OXA by MTT assay in control cells (siNTC) and CXCL8 or CXCL1 knockdown cells (siCXCL8 or siCXCL1, respectively). Forty-eight hours after gene silencing, cells were seeded in 96-well plates and treated for 24 h with doses of OXA ranging 0–100 μM. CXCL8 and CXCL1 knockdown efficiency was ~90% and ~60% at mRNA level, respectively (insets on Fig. 7a,b). Under these conditions, we found that CXCL8 and CXCL1 gene silencing in HTOXAR3 led to a 35% and a 20% decrease in OXA IC50 value as compared to siNTC cells, respectively (Fig. 7a,b).

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