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Cyanidin-3-glucoside inhibits ethanol-induced invasion of breast cancer cells overexpressing ErbB2.

Xu M, Bower KA, Wang S, Frank JA, Chen G, Ding M, Wang S, Shi X, Ke Z, Luo J - Mol. Cancer (2010)

Bottom Line: C3G decreased ethanol-mediated cell adhesion to the extracellular matrix (ECM) as well as the amount of focal adhesions and the formation of lamellipodial protrusion.It inhibited ethanol-stimulated phosphorylation of ErbB2, cSrc, FAK and p130(Cas), as well as interactions among these proteins.C3G may be beneficial in preventing/reducing ethanol-induced breast cancer metastasis.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Internal Medicine, University of Kentucky College of Medicine, Lexington, KY 40536, USA.

ABSTRACT

Background: Ethanol is a tumor promoter. Both epidemiological and experimental studies suggest that ethanol may enhance the metastasis of breast cancer cells. We have previously demonstrated that ethanol increased the migration/invasion of breast cancer cells expressing high levels of ErbB2. Amplification of ErbB2 is found in 20-30% of breast cancer patients and is associated with poor prognosis. We sought to identify agents that can prevent or ameliorate ethanol-induced invasion of breast cancer cells. Cyanidin-3-glucoside (C3G), an anthocyanin present in many vegetables and fruits, is a potent natural antioxidant. Ethanol exposure causes the accumulation of intracellular reactive oxygen species (ROS). This study evaluated the effect of C3G on ethanol-induced breast cancer cell migration/invasion.

Results: C3G attenuated ethanol-induced migration/invasion of breast cancer cells expressing high levels of ErbB2 (BT474, MDA-MB231 and MCF7(ErbB2)) in a concentration dependent manner. C3G decreased ethanol-mediated cell adhesion to the extracellular matrix (ECM) as well as the amount of focal adhesions and the formation of lamellipodial protrusion. It inhibited ethanol-stimulated phosphorylation of ErbB2, cSrc, FAK and p130(Cas), as well as interactions among these proteins. C3G abolished ethanol-mediated p130(Cas)/JNK interaction.

Conclusions: C3G blocks ethanol-induced activation of the ErbB2/cSrc/FAK pathway which is necessary for cell migration/invasion. C3G may be beneficial in preventing/reducing ethanol-induced breast cancer metastasis.

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Effects of C3G on ethanol-mediated formation of focal adhesions. MDA-MB231 cells were treated with ethanol (0 or 400 mg/dl) with/without C3G (40 μM) for 48 h. Cells were seeded on fibronectin-coated coverslips for 3 h. A: The expression of paxillin (Alexa Fluor 488) and phosphorylated FAK (Tyr861) (Alexa Fluor 594) were detected by immunofluorescent staining. Arrows indicate the co-localization of p-FAK (Tyr861) and paxillin. Scale bar = 5 μm. B: Focal adhesions were counted randomly on 10 or more cells. The number of focal adhesions per cell was calculated. Each datum point was the mean ± SEM of three independent experiments. * denotes a significant difference from untreated controls. # denotes a significant difference from ethanol-treated groups.
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Figure 7: Effects of C3G on ethanol-mediated formation of focal adhesions. MDA-MB231 cells were treated with ethanol (0 or 400 mg/dl) with/without C3G (40 μM) for 48 h. Cells were seeded on fibronectin-coated coverslips for 3 h. A: The expression of paxillin (Alexa Fluor 488) and phosphorylated FAK (Tyr861) (Alexa Fluor 594) were detected by immunofluorescent staining. Arrows indicate the co-localization of p-FAK (Tyr861) and paxillin. Scale bar = 5 μm. B: Focal adhesions were counted randomly on 10 or more cells. The number of focal adhesions per cell was calculated. Each datum point was the mean ± SEM of three independent experiments. * denotes a significant difference from untreated controls. # denotes a significant difference from ethanol-treated groups.

Mentions: The initiation of cell migration requires the development of membrane protrusion, the lamellipodium and the assembly of dynamic focal adhesions with the ECM [29]. We sought to determine whether C3G affected the formation of the lamellipodium and focal adhesions. We used MDA-MB231 cells for this experiment because these cells displayed more prominent lamellipodium and focal adhesions during the migration process. Figure 6A shows that actin filament distribution was concentrated at the leading edge/lamellipodia in ethanol-treated MDA-MB231 cells. Ethanol caused an approximate 3-fold increase in the number of lamellipodia (Figure 6B). C3G inhibited ethanol-induced lamellipodia formation; however, the inhibition was not concentration-dependent and C3G at 10 or 40 μM had a similar effect (Figure 6B). We demonstrated an accumulation of p-FAK(Tyr861) at the leading area in ethanol-treated cells (Figures 6A and 7A). Ethanol also caused redistribution of paxillin, and more paxillin was localized at the leading edge following ethanol exposure (Figure 7A). Since the activation of FAK leads to the recruitment of paxillin and p130Cas to focal adhesion sites [27,30], we examined the effect of ethanol on focal adhesions. Ethanol enhanced the assembly of focal adhesions and C3G significantly inhibited ethanol-induced formation of focal adhesions (Figure 7B).


Cyanidin-3-glucoside inhibits ethanol-induced invasion of breast cancer cells overexpressing ErbB2.

Xu M, Bower KA, Wang S, Frank JA, Chen G, Ding M, Wang S, Shi X, Ke Z, Luo J - Mol. Cancer (2010)

Effects of C3G on ethanol-mediated formation of focal adhesions. MDA-MB231 cells were treated with ethanol (0 or 400 mg/dl) with/without C3G (40 μM) for 48 h. Cells were seeded on fibronectin-coated coverslips for 3 h. A: The expression of paxillin (Alexa Fluor 488) and phosphorylated FAK (Tyr861) (Alexa Fluor 594) were detected by immunofluorescent staining. Arrows indicate the co-localization of p-FAK (Tyr861) and paxillin. Scale bar = 5 μm. B: Focal adhesions were counted randomly on 10 or more cells. The number of focal adhesions per cell was calculated. Each datum point was the mean ± SEM of three independent experiments. * denotes a significant difference from untreated controls. # denotes a significant difference from ethanol-treated groups.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Figure 7: Effects of C3G on ethanol-mediated formation of focal adhesions. MDA-MB231 cells were treated with ethanol (0 or 400 mg/dl) with/without C3G (40 μM) for 48 h. Cells were seeded on fibronectin-coated coverslips for 3 h. A: The expression of paxillin (Alexa Fluor 488) and phosphorylated FAK (Tyr861) (Alexa Fluor 594) were detected by immunofluorescent staining. Arrows indicate the co-localization of p-FAK (Tyr861) and paxillin. Scale bar = 5 μm. B: Focal adhesions were counted randomly on 10 or more cells. The number of focal adhesions per cell was calculated. Each datum point was the mean ± SEM of three independent experiments. * denotes a significant difference from untreated controls. # denotes a significant difference from ethanol-treated groups.
Mentions: The initiation of cell migration requires the development of membrane protrusion, the lamellipodium and the assembly of dynamic focal adhesions with the ECM [29]. We sought to determine whether C3G affected the formation of the lamellipodium and focal adhesions. We used MDA-MB231 cells for this experiment because these cells displayed more prominent lamellipodium and focal adhesions during the migration process. Figure 6A shows that actin filament distribution was concentrated at the leading edge/lamellipodia in ethanol-treated MDA-MB231 cells. Ethanol caused an approximate 3-fold increase in the number of lamellipodia (Figure 6B). C3G inhibited ethanol-induced lamellipodia formation; however, the inhibition was not concentration-dependent and C3G at 10 or 40 μM had a similar effect (Figure 6B). We demonstrated an accumulation of p-FAK(Tyr861) at the leading area in ethanol-treated cells (Figures 6A and 7A). Ethanol also caused redistribution of paxillin, and more paxillin was localized at the leading edge following ethanol exposure (Figure 7A). Since the activation of FAK leads to the recruitment of paxillin and p130Cas to focal adhesion sites [27,30], we examined the effect of ethanol on focal adhesions. Ethanol enhanced the assembly of focal adhesions and C3G significantly inhibited ethanol-induced formation of focal adhesions (Figure 7B).

Bottom Line: C3G decreased ethanol-mediated cell adhesion to the extracellular matrix (ECM) as well as the amount of focal adhesions and the formation of lamellipodial protrusion.It inhibited ethanol-stimulated phosphorylation of ErbB2, cSrc, FAK and p130(Cas), as well as interactions among these proteins.C3G may be beneficial in preventing/reducing ethanol-induced breast cancer metastasis.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Internal Medicine, University of Kentucky College of Medicine, Lexington, KY 40536, USA.

ABSTRACT

Background: Ethanol is a tumor promoter. Both epidemiological and experimental studies suggest that ethanol may enhance the metastasis of breast cancer cells. We have previously demonstrated that ethanol increased the migration/invasion of breast cancer cells expressing high levels of ErbB2. Amplification of ErbB2 is found in 20-30% of breast cancer patients and is associated with poor prognosis. We sought to identify agents that can prevent or ameliorate ethanol-induced invasion of breast cancer cells. Cyanidin-3-glucoside (C3G), an anthocyanin present in many vegetables and fruits, is a potent natural antioxidant. Ethanol exposure causes the accumulation of intracellular reactive oxygen species (ROS). This study evaluated the effect of C3G on ethanol-induced breast cancer cell migration/invasion.

Results: C3G attenuated ethanol-induced migration/invasion of breast cancer cells expressing high levels of ErbB2 (BT474, MDA-MB231 and MCF7(ErbB2)) in a concentration dependent manner. C3G decreased ethanol-mediated cell adhesion to the extracellular matrix (ECM) as well as the amount of focal adhesions and the formation of lamellipodial protrusion. It inhibited ethanol-stimulated phosphorylation of ErbB2, cSrc, FAK and p130(Cas), as well as interactions among these proteins. C3G abolished ethanol-mediated p130(Cas)/JNK interaction.

Conclusions: C3G blocks ethanol-induced activation of the ErbB2/cSrc/FAK pathway which is necessary for cell migration/invasion. C3G may be beneficial in preventing/reducing ethanol-induced breast cancer metastasis.

Show MeSH
Related in: MedlinePlus