Limits...
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

Combination of OXA and Curcumin in OXA-sensitive and –resistant CRC cell lines.(a) Graphic representation of the Combination Index (CI) values corresponding to different OXA plus Curcumin (CURC) treatment schedules in the HTOXAR3 cell line. (b) Bar graphs representing mean ± SEM percentage of cell viability after a 24-hour treatment with OXA, Curcumin or their concomitant combination at the indicated doses in HT29 and HTOXAR3 cells (c). *p-value < 0.05; **p-value < 0.01; ***p-value < 0.001 relative to the indicated treatment condition. (d) OXA doses (mean ± SEM) corresponding to indicated inhibitory concentrations (IC) in HT29 and HTOXAR3 cells as a single agent or when combined with Curcumin concomitantly for 24 h. (e) OXA and Curcumin doses (mean ± SD) corresponding to the indicated inhibitory concentrations (IC) when given as single agents or in a 24 h-concomitant schedule in HT29 and HTOXAR3. CI represents the combination index values in each case. Synergistic values (CI < 1) are highlighted in green. (f) Bar graph representing the percentage (mean ± SEM) of colonies in HT29 and HTOXAR3 cells after 24 h of the indicated treatments. *p-value < 0.05; **p-value < 0.01; ***p-value < 0.001 relative to NT (Non-treated cells). #p-value < 0.05 relative to OXA individual treatment. *p-value < 0.05 as compared to HT29. (g) Bar graph representing the percentage (mean ± SEM) of dead cells after 24, 48 and 72 h of treatment with OXA, Curcumin or their combination (as compared to non-treated controls) at their corresponding IC50 doses in HT29 and HTOXAR3 cell lines. *p-value < 0.05; **p-value < 0.01, relative to NT. #p-value < 0,05, relative to Curcumin individual treatment.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4835769&req=5

f3: Combination of OXA and Curcumin in OXA-sensitive and –resistant CRC cell lines.(a) Graphic representation of the Combination Index (CI) values corresponding to different OXA plus Curcumin (CURC) treatment schedules in the HTOXAR3 cell line. (b) Bar graphs representing mean ± SEM percentage of cell viability after a 24-hour treatment with OXA, Curcumin or their concomitant combination at the indicated doses in HT29 and HTOXAR3 cells (c). *p-value < 0.05; **p-value < 0.01; ***p-value < 0.001 relative to the indicated treatment condition. (d) OXA doses (mean ± SEM) corresponding to indicated inhibitory concentrations (IC) in HT29 and HTOXAR3 cells as a single agent or when combined with Curcumin concomitantly for 24 h. (e) OXA and Curcumin doses (mean ± SD) corresponding to the indicated inhibitory concentrations (IC) when given as single agents or in a 24 h-concomitant schedule in HT29 and HTOXAR3. CI represents the combination index values in each case. Synergistic values (CI < 1) are highlighted in green. (f) Bar graph representing the percentage (mean ± SEM) of colonies in HT29 and HTOXAR3 cells after 24 h of the indicated treatments. *p-value < 0.05; **p-value < 0.01; ***p-value < 0.001 relative to NT (Non-treated cells). #p-value < 0.05 relative to OXA individual treatment. *p-value < 0.05 as compared to HT29. (g) Bar graph representing the percentage (mean ± SEM) of dead cells after 24, 48 and 72 h of treatment with OXA, Curcumin or their combination (as compared to non-treated controls) at their corresponding IC50 doses in HT29 and HTOXAR3 cell lines. *p-value < 0.05; **p-value < 0.01, relative to NT. #p-value < 0,05, relative to Curcumin individual treatment.

Mentions: A panel of 3 CRC cell lines and their respective OXA-resistant cells were treated with increasing concentrations of Curcumin for 24 h. Curcumin decreased cell proliferation of all cell lines in a dose-dependent manner (IC50 range 9.5–14.6 μM). Importantly, HTOXAR3 and LoVOXAR3 (Supplementary Fig. S2a,b) cells were more resistant to Curcumin treatment as compared with HT29 and LoVo cell lines whereas DLDOXAR3 cells were more sensitive than their parental sensitive cell line, DLD1 (Supplementary Fig. S2c). Next we wanted to determine the most synergistic combination between OXA and Curcumin. To do that, HTOXAR3 resistant cells were treated with different combination schedules, with the most synergistic being the concomitant treatment of both drugs for 24 h, where the CI was <1 from IC40 onwards, CI = 1 at IC30 and was slightly higher than 1 at very low doses (IC10 and 20) (Fig. 3a). Therefore, we decided to use this combination in order to evaluate the effect of Curcumin on the reversion of OXA-acquired resistance in our models. We observed that this schedule significantly decreased cell proliferation of both HT29 and HTOXAR3 cell lines (Fig. 3b,c) as compared with individual treatments. Remarkably, this reduction of cell viability was more evident in the HTOXAR3 cell line where in fact the combined treatment was able to revert the OXA-acquired resistance phenotype leading to a 65% reduction in the OXA IC50 (10.6 ± 2.2 μM with the combination vs. 30.2 ± 4.2 μM in individual treatment) and brought it closer to that of the HT29 sensitive cell line (8.45 ± 1.6 μM) (Fig. 3d). While in the HT29 cell line, the combination of Curcumin plus OXA was additive at the IC50 of these drugs and synergistic at doses higher than the IC50, in the resistant HTOXAR3 cells, the treatment started to be synergistic at the IC50 (Fig. 3e). Clonogenic assays revealed that while both drugs reduced the number of colonies as compared with untreated cells, their combination led to a complete absence of colonies in both HT29 and HTOXAR3 cells (Fig. 3f). In order to determine if these results could be explained by an increased cell death after the combined treatment, HT29 and HTOXAR3 cells were treated with OXA, Curcumin or their combination at their respective IC50 doses for 24, 48 and 72 h. As can be seen in Fig. 3g, OXA induced cell death rates below 25% after 72 h of treatment in both cell lines while in the case of Curcumin alone these rates were less than 10% in all cases. However the percentage of dead cells was increased after treatment with OXA plus Curcumin for 72 h in HT29 cells and for 48 and 72 h in HTOXAR3 only as compared with Curcumin alone. Similar results were obtained in the LoVo/LoVOXAR3 pair although an additive effect was observed only at high doses (IC90) of the two drugs in both cell lines (Supplementary Fig. S3).


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)

Combination of OXA and Curcumin in OXA-sensitive and –resistant CRC cell lines.(a) Graphic representation of the Combination Index (CI) values corresponding to different OXA plus Curcumin (CURC) treatment schedules in the HTOXAR3 cell line. (b) Bar graphs representing mean ± SEM percentage of cell viability after a 24-hour treatment with OXA, Curcumin or their concomitant combination at the indicated doses in HT29 and HTOXAR3 cells (c). *p-value < 0.05; **p-value < 0.01; ***p-value < 0.001 relative to the indicated treatment condition. (d) OXA doses (mean ± SEM) corresponding to indicated inhibitory concentrations (IC) in HT29 and HTOXAR3 cells as a single agent or when combined with Curcumin concomitantly for 24 h. (e) OXA and Curcumin doses (mean ± SD) corresponding to the indicated inhibitory concentrations (IC) when given as single agents or in a 24 h-concomitant schedule in HT29 and HTOXAR3. CI represents the combination index values in each case. Synergistic values (CI < 1) are highlighted in green. (f) Bar graph representing the percentage (mean ± SEM) of colonies in HT29 and HTOXAR3 cells after 24 h of the indicated treatments. *p-value < 0.05; **p-value < 0.01; ***p-value < 0.001 relative to NT (Non-treated cells). #p-value < 0.05 relative to OXA individual treatment. *p-value < 0.05 as compared to HT29. (g) Bar graph representing the percentage (mean ± SEM) of dead cells after 24, 48 and 72 h of treatment with OXA, Curcumin or their combination (as compared to non-treated controls) at their corresponding IC50 doses in HT29 and HTOXAR3 cell lines. *p-value < 0.05; **p-value < 0.01, relative to NT. #p-value < 0,05, relative to Curcumin individual treatment.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f3: Combination of OXA and Curcumin in OXA-sensitive and –resistant CRC cell lines.(a) Graphic representation of the Combination Index (CI) values corresponding to different OXA plus Curcumin (CURC) treatment schedules in the HTOXAR3 cell line. (b) Bar graphs representing mean ± SEM percentage of cell viability after a 24-hour treatment with OXA, Curcumin or their concomitant combination at the indicated doses in HT29 and HTOXAR3 cells (c). *p-value < 0.05; **p-value < 0.01; ***p-value < 0.001 relative to the indicated treatment condition. (d) OXA doses (mean ± SEM) corresponding to indicated inhibitory concentrations (IC) in HT29 and HTOXAR3 cells as a single agent or when combined with Curcumin concomitantly for 24 h. (e) OXA and Curcumin doses (mean ± SD) corresponding to the indicated inhibitory concentrations (IC) when given as single agents or in a 24 h-concomitant schedule in HT29 and HTOXAR3. CI represents the combination index values in each case. Synergistic values (CI < 1) are highlighted in green. (f) Bar graph representing the percentage (mean ± SEM) of colonies in HT29 and HTOXAR3 cells after 24 h of the indicated treatments. *p-value < 0.05; **p-value < 0.01; ***p-value < 0.001 relative to NT (Non-treated cells). #p-value < 0.05 relative to OXA individual treatment. *p-value < 0.05 as compared to HT29. (g) Bar graph representing the percentage (mean ± SEM) of dead cells after 24, 48 and 72 h of treatment with OXA, Curcumin or their combination (as compared to non-treated controls) at their corresponding IC50 doses in HT29 and HTOXAR3 cell lines. *p-value < 0.05; **p-value < 0.01, relative to NT. #p-value < 0,05, relative to Curcumin individual treatment.
Mentions: A panel of 3 CRC cell lines and their respective OXA-resistant cells were treated with increasing concentrations of Curcumin for 24 h. Curcumin decreased cell proliferation of all cell lines in a dose-dependent manner (IC50 range 9.5–14.6 μM). Importantly, HTOXAR3 and LoVOXAR3 (Supplementary Fig. S2a,b) cells were more resistant to Curcumin treatment as compared with HT29 and LoVo cell lines whereas DLDOXAR3 cells were more sensitive than their parental sensitive cell line, DLD1 (Supplementary Fig. S2c). Next we wanted to determine the most synergistic combination between OXA and Curcumin. To do that, HTOXAR3 resistant cells were treated with different combination schedules, with the most synergistic being the concomitant treatment of both drugs for 24 h, where the CI was <1 from IC40 onwards, CI = 1 at IC30 and was slightly higher than 1 at very low doses (IC10 and 20) (Fig. 3a). Therefore, we decided to use this combination in order to evaluate the effect of Curcumin on the reversion of OXA-acquired resistance in our models. We observed that this schedule significantly decreased cell proliferation of both HT29 and HTOXAR3 cell lines (Fig. 3b,c) as compared with individual treatments. Remarkably, this reduction of cell viability was more evident in the HTOXAR3 cell line where in fact the combined treatment was able to revert the OXA-acquired resistance phenotype leading to a 65% reduction in the OXA IC50 (10.6 ± 2.2 μM with the combination vs. 30.2 ± 4.2 μM in individual treatment) and brought it closer to that of the HT29 sensitive cell line (8.45 ± 1.6 μM) (Fig. 3d). While in the HT29 cell line, the combination of Curcumin plus OXA was additive at the IC50 of these drugs and synergistic at doses higher than the IC50, in the resistant HTOXAR3 cells, the treatment started to be synergistic at the IC50 (Fig. 3e). Clonogenic assays revealed that while both drugs reduced the number of colonies as compared with untreated cells, their combination led to a complete absence of colonies in both HT29 and HTOXAR3 cells (Fig. 3f). In order to determine if these results could be explained by an increased cell death after the combined treatment, HT29 and HTOXAR3 cells were treated with OXA, Curcumin or their combination at their respective IC50 doses for 24, 48 and 72 h. As can be seen in Fig. 3g, OXA induced cell death rates below 25% after 72 h of treatment in both cell lines while in the case of Curcumin alone these rates were less than 10% in all cases. However the percentage of dead cells was increased after treatment with OXA plus Curcumin for 72 h in HT29 cells and for 48 and 72 h in HTOXAR3 only as compared with Curcumin alone. Similar results were obtained in the LoVo/LoVOXAR3 pair although an additive effect was observed only at high doses (IC90) of the two drugs in both cell lines (Supplementary Fig. S3).

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