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Low molecular weight procyanidins from grape seeds enhance the impact of 5-Fluorouracil chemotherapy on Caco-2 human colon cancer cells.

Cheah KY, Howarth GS, Bindon KA, Kennedy JA, Bastian SE - PLoS ONE (2014)

Bottom Line: All isolated fractions significantly reduced Caco-2 cell viability compared to the control (P<0.05), but F2 and F3 (mDP 2-6) were the most active fractions (immature F2 = 32% mDP 2.4, F3 = 35% mDP 5.8 and mature F2 = 13% mDP 3.6 and F3 = 17% mDP 5.9; percentage of viable cells remaining) on Caco-2 cells.When combined with 5-FU, immature fractions F1-F3 enhanced the cell toxicity effects of 5-FU by 27-73% (P<0.05).Moreover, some fractions alone were more potent at decreasing viability in Caco-2 cells (P<0.05; immature fractions = 65-68% and mature fractions = 83-87%) compared to 5-FU alone (37%).

View Article: PubMed Central - PubMed

Affiliation: Wine Science and Business Group, School of Agriculture, Food and Wine, Waite Campus, The University of Adelaide, PMB 1, Glen Osmond, Australia; Centre for Paediatric and Adolescent Gastroenterology, Children, Youth and Women's Health Service, North Adelaide, Australia.

ABSTRACT

Objective: Grape seed procyanidins (PC) are flavan-3-ol oligomers and polymers known for their biological activity in the gut. Grape seed extract (GSE) have been reported to reduce intestinal injury in a rat model of mucositis. We sought to investigate effects of purified PC fractions differing in mean degree of polymerization (mDP) combined with 5-Fluorouracil (5-FU) chemotherapy on the viability of colon cancer cells (Caco-2).

Design: SixPC fractions (F1-F6) were isolated from Cabernet Sauvignon seeds at two ripeness stages: pre-veraison unripe (immature) and ripe (mature), utilizing step gradient, low-pressure chromatography on a Sephadex LH-20 resin. Fractions were tested on Caco-2 cells, alone and in combination with 5-FU. Eluted fractions were characterized by phloroglucinolysis and gel permeation chromatography. Cell viability was determined by the 3-(4,5-Dimethylthiazol-2yl)-2,5-diphenyl-tetrazolium bromide) (MTT) assay.

Results: All isolated fractions significantly reduced Caco-2 cell viability compared to the control (P<0.05), but F2 and F3 (mDP 2-6) were the most active fractions (immature F2 = 32% mDP 2.4, F3 = 35% mDP 5.8 and mature F2 = 13% mDP 3.6 and F3 = 17% mDP 5.9; percentage of viable cells remaining) on Caco-2 cells. When combined with 5-FU, immature fractions F1-F3 enhanced the cell toxicity effects of 5-FU by 27-73% (P<0.05). Mature seed PC fractions (F1-F4) significantly enhanced the toxicity of 5-FU by 60-83% against Caco-2 cells (P<0.05). Moreover, some fractions alone were more potent at decreasing viability in Caco-2 cells (P<0.05; immature fractions = 65-68% and mature fractions = 83-87%) compared to 5-FU alone (37%).

Conclusions: PCs of mDP 2-6 (immature F1-F3 and mature F1 and F4)not only enhanced the impact of 5-FU in killing Caco-2 cells, but also surpassed standard 5-FU chemotherapy as an anti-cancer agent.The bioactivity of PC is therefore attributed primarily to lower molecular weight PCs.

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Viability of Caco-2 cells after exposure to fractions from immature (A) and mature (B) seed extracts for 72 hr measured by 3-(4,5-Dimethylthiazol-2yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay.Data are expressed as IC50 or dose (µg/mL) inhibiting cell viability to 50% (mean ± SEM) of 3 independent experiments. Statistical analysis was determined by one-way ANOVA with Tukey'spost-hoc test. Values with different letters (a,b,c) in each column were statistically different at P<0.05.
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pone-0098921-g002: Viability of Caco-2 cells after exposure to fractions from immature (A) and mature (B) seed extracts for 72 hr measured by 3-(4,5-Dimethylthiazol-2yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay.Data are expressed as IC50 or dose (µg/mL) inhibiting cell viability to 50% (mean ± SEM) of 3 independent experiments. Statistical analysis was determined by one-way ANOVA with Tukey'spost-hoc test. Values with different letters (a,b,c) in each column were statistically different at P<0.05.

Mentions: The cytotoxic effects of seed fractions on Caco-2 cells were determined by the MTT assay (Figure 2). Caco-2 cells were exposed to seed fractions for 72 hr and the data expressed as IC50, defined as the dose of each compound that inhibited cell viability to 50%, representing the mean value of three independent experiments. All fractions showed a degree of toxicity as indicated by decreased absorbance values. The IC50value (mean of the three independent experiments) for immature seed fractions (F1-F6) ranged from 17.7–70.2 µg/mL (Figure 2A). The number of viable cells was positively correlated with the mDP of immature seed fractions (r2 = 0.48, P<0.05) i.e., the fractions with highest mDP (F5 and F6) were less toxic to Caco-2 cells. Similar trends in terms of cytotoxic effects were also observed for the application of mature seed fractions (Figure 2B), where the number of viable cells was similarly correlated with the mDP of the fractions (r2 = 0.50, P<0.05). Mature seed fractions exhibited stronger cytotoxicity compared to immature seed fractions (Figure 2A and 2B).


Low molecular weight procyanidins from grape seeds enhance the impact of 5-Fluorouracil chemotherapy on Caco-2 human colon cancer cells.

Cheah KY, Howarth GS, Bindon KA, Kennedy JA, Bastian SE - PLoS ONE (2014)

Viability of Caco-2 cells after exposure to fractions from immature (A) and mature (B) seed extracts for 72 hr measured by 3-(4,5-Dimethylthiazol-2yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay.Data are expressed as IC50 or dose (µg/mL) inhibiting cell viability to 50% (mean ± SEM) of 3 independent experiments. Statistical analysis was determined by one-way ANOVA with Tukey'spost-hoc test. Values with different letters (a,b,c) in each column were statistically different at P<0.05.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4048230&req=5

pone-0098921-g002: Viability of Caco-2 cells after exposure to fractions from immature (A) and mature (B) seed extracts for 72 hr measured by 3-(4,5-Dimethylthiazol-2yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay.Data are expressed as IC50 or dose (µg/mL) inhibiting cell viability to 50% (mean ± SEM) of 3 independent experiments. Statistical analysis was determined by one-way ANOVA with Tukey'spost-hoc test. Values with different letters (a,b,c) in each column were statistically different at P<0.05.
Mentions: The cytotoxic effects of seed fractions on Caco-2 cells were determined by the MTT assay (Figure 2). Caco-2 cells were exposed to seed fractions for 72 hr and the data expressed as IC50, defined as the dose of each compound that inhibited cell viability to 50%, representing the mean value of three independent experiments. All fractions showed a degree of toxicity as indicated by decreased absorbance values. The IC50value (mean of the three independent experiments) for immature seed fractions (F1-F6) ranged from 17.7–70.2 µg/mL (Figure 2A). The number of viable cells was positively correlated with the mDP of immature seed fractions (r2 = 0.48, P<0.05) i.e., the fractions with highest mDP (F5 and F6) were less toxic to Caco-2 cells. Similar trends in terms of cytotoxic effects were also observed for the application of mature seed fractions (Figure 2B), where the number of viable cells was similarly correlated with the mDP of the fractions (r2 = 0.50, P<0.05). Mature seed fractions exhibited stronger cytotoxicity compared to immature seed fractions (Figure 2A and 2B).

Bottom Line: All isolated fractions significantly reduced Caco-2 cell viability compared to the control (P<0.05), but F2 and F3 (mDP 2-6) were the most active fractions (immature F2 = 32% mDP 2.4, F3 = 35% mDP 5.8 and mature F2 = 13% mDP 3.6 and F3 = 17% mDP 5.9; percentage of viable cells remaining) on Caco-2 cells.When combined with 5-FU, immature fractions F1-F3 enhanced the cell toxicity effects of 5-FU by 27-73% (P<0.05).Moreover, some fractions alone were more potent at decreasing viability in Caco-2 cells (P<0.05; immature fractions = 65-68% and mature fractions = 83-87%) compared to 5-FU alone (37%).

View Article: PubMed Central - PubMed

Affiliation: Wine Science and Business Group, School of Agriculture, Food and Wine, Waite Campus, The University of Adelaide, PMB 1, Glen Osmond, Australia; Centre for Paediatric and Adolescent Gastroenterology, Children, Youth and Women's Health Service, North Adelaide, Australia.

ABSTRACT

Objective: Grape seed procyanidins (PC) are flavan-3-ol oligomers and polymers known for their biological activity in the gut. Grape seed extract (GSE) have been reported to reduce intestinal injury in a rat model of mucositis. We sought to investigate effects of purified PC fractions differing in mean degree of polymerization (mDP) combined with 5-Fluorouracil (5-FU) chemotherapy on the viability of colon cancer cells (Caco-2).

Design: SixPC fractions (F1-F6) were isolated from Cabernet Sauvignon seeds at two ripeness stages: pre-veraison unripe (immature) and ripe (mature), utilizing step gradient, low-pressure chromatography on a Sephadex LH-20 resin. Fractions were tested on Caco-2 cells, alone and in combination with 5-FU. Eluted fractions were characterized by phloroglucinolysis and gel permeation chromatography. Cell viability was determined by the 3-(4,5-Dimethylthiazol-2yl)-2,5-diphenyl-tetrazolium bromide) (MTT) assay.

Results: All isolated fractions significantly reduced Caco-2 cell viability compared to the control (P<0.05), but F2 and F3 (mDP 2-6) were the most active fractions (immature F2 = 32% mDP 2.4, F3 = 35% mDP 5.8 and mature F2 = 13% mDP 3.6 and F3 = 17% mDP 5.9; percentage of viable cells remaining) on Caco-2 cells. When combined with 5-FU, immature fractions F1-F3 enhanced the cell toxicity effects of 5-FU by 27-73% (P<0.05). Mature seed PC fractions (F1-F4) significantly enhanced the toxicity of 5-FU by 60-83% against Caco-2 cells (P<0.05). Moreover, some fractions alone were more potent at decreasing viability in Caco-2 cells (P<0.05; immature fractions = 65-68% and mature fractions = 83-87%) compared to 5-FU alone (37%).

Conclusions: PCs of mDP 2-6 (immature F1-F3 and mature F1 and F4)not only enhanced the impact of 5-FU in killing Caco-2 cells, but also surpassed standard 5-FU chemotherapy as an anti-cancer agent.The bioactivity of PC is therefore attributed primarily to lower molecular weight PCs.

Show MeSH
Related in: MedlinePlus