Limits...
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.

Show MeSH

Related in: MedlinePlus

Effects of C3G and antioxidants on ethanol-induced ROS generation, cell invasion and ErbB2 phosphorylation. A: MCF7ErbB2 cells were exposed to ethanol (0 or 400 mg/dl) with/without C3G (10 μM), SOD (50 U/ml)/catalase (200 U/ml), NAC (5 mM) or vitamin C (20 μM) for 48 h. Intracellular ROS levels were measured by flow cytometry as described under the Materials and Methods. B: The invasive potential of MCF7ErbB2 cells was evaluated as described above and expressed relative to untreated controls. C: The phosphorylation of ErbB2 in MCF7ErbB2 cells was analyzed with immunoblotting. The experiment was replicated three times. * denotes a statistically significant difference from untreated controls. # denotes a significant difference from ethanol-treated groups. # denotes a significant difference from ethanol- and C3G-treated groups.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2984473&req=5

Figure 8: Effects of C3G and antioxidants on ethanol-induced ROS generation, cell invasion and ErbB2 phosphorylation. A: MCF7ErbB2 cells were exposed to ethanol (0 or 400 mg/dl) with/without C3G (10 μM), SOD (50 U/ml)/catalase (200 U/ml), NAC (5 mM) or vitamin C (20 μM) for 48 h. Intracellular ROS levels were measured by flow cytometry as described under the Materials and Methods. B: The invasive potential of MCF7ErbB2 cells was evaluated as described above and expressed relative to untreated controls. C: The phosphorylation of ErbB2 in MCF7ErbB2 cells was analyzed with immunoblotting. The experiment was replicated three times. * denotes a statistically significant difference from untreated controls. # denotes a significant difference from ethanol-treated groups. # denotes a significant difference from ethanol- and C3G-treated groups.

Mentions: Ethanol causes intracellular accumulation of reactive oxygen species (ROS) and induces oxidative stress [10,31]. Since C3G is a potent antioxidant, the inhibitory effect of C3G on ethanol-induced migration/invasion may be mediated by its antioxidant property. We evaluated the effect of other antioxidants at concentrations that had a similar ROS scavenging capacity as C3G. Superoxide dismutase (SOD) is a scavenger for O2• and catalase is a scavenger for hydrogen peroxide (H2O2). N-aceytlcysteine (NAC) (5 mM), vitamin C (20 μM) and SOD (50 U/ml) plus catalase (200 U/ml) had approximately the same antioxidant effect as C3G (10 μM) (Figure 8A). As shown in Figure 8B, C3G most effectively inhibited ethanol-enhanced invasion of breast cancer cells; NAC and vitamin C also provided significant inhibition, but to a lesser extent. On the other hand, SOD plus catalase had little effect on ethanol-enhanced cell invasion. A similar result regarding the effect of C3G and other antioxidants on ethanol-induced ErbB2 phosphorylation was observed (Figure 8C).


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 and antioxidants on ethanol-induced ROS generation, cell invasion and ErbB2 phosphorylation. A: MCF7ErbB2 cells were exposed to ethanol (0 or 400 mg/dl) with/without C3G (10 μM), SOD (50 U/ml)/catalase (200 U/ml), NAC (5 mM) or vitamin C (20 μM) for 48 h. Intracellular ROS levels were measured by flow cytometry as described under the Materials and Methods. B: The invasive potential of MCF7ErbB2 cells was evaluated as described above and expressed relative to untreated controls. C: The phosphorylation of ErbB2 in MCF7ErbB2 cells was analyzed with immunoblotting. The experiment was replicated three times. * denotes a statistically significant difference from untreated controls. # denotes a significant difference from ethanol-treated groups. # denotes a significant difference from ethanol- and C3G-treated groups.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2984473&req=5

Figure 8: Effects of C3G and antioxidants on ethanol-induced ROS generation, cell invasion and ErbB2 phosphorylation. A: MCF7ErbB2 cells were exposed to ethanol (0 or 400 mg/dl) with/without C3G (10 μM), SOD (50 U/ml)/catalase (200 U/ml), NAC (5 mM) or vitamin C (20 μM) for 48 h. Intracellular ROS levels were measured by flow cytometry as described under the Materials and Methods. B: The invasive potential of MCF7ErbB2 cells was evaluated as described above and expressed relative to untreated controls. C: The phosphorylation of ErbB2 in MCF7ErbB2 cells was analyzed with immunoblotting. The experiment was replicated three times. * denotes a statistically significant difference from untreated controls. # denotes a significant difference from ethanol-treated groups. # denotes a significant difference from ethanol- and C3G-treated groups.
Mentions: Ethanol causes intracellular accumulation of reactive oxygen species (ROS) and induces oxidative stress [10,31]. Since C3G is a potent antioxidant, the inhibitory effect of C3G on ethanol-induced migration/invasion may be mediated by its antioxidant property. We evaluated the effect of other antioxidants at concentrations that had a similar ROS scavenging capacity as C3G. Superoxide dismutase (SOD) is a scavenger for O2• and catalase is a scavenger for hydrogen peroxide (H2O2). N-aceytlcysteine (NAC) (5 mM), vitamin C (20 μM) and SOD (50 U/ml) plus catalase (200 U/ml) had approximately the same antioxidant effect as C3G (10 μM) (Figure 8A). As shown in Figure 8B, C3G most effectively inhibited ethanol-enhanced invasion of breast cancer cells; NAC and vitamin C also provided significant inhibition, but to a lesser extent. On the other hand, SOD plus catalase had little effect on ethanol-enhanced cell invasion. A similar result regarding the effect of C3G and other antioxidants on ethanol-induced ErbB2 phosphorylation was observed (Figure 8C).

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