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Glaucine inhibits breast cancer cell migration and invasion by inhibiting MMP-9 gene expression through the suppression of NF-κB activation.

Kang H, Jang SW, Pak JH, Shim S - Mol. Cell. Biochem. (2015)

Bottom Line: We further show that glaucine significantly blocks phorbol 12-myristate 13-acetate (PMA)-induced MMP-9 expression and activity in a dose-dependent manner.Moreover, glaucine attenuates PMA-induced IκBα degradation and nuclear translocation of NF-κB.Taken together, our findings indicate that the MMP-9 inhibitory activity of glaucine and its abilities to attenuate IκBα and NF-κB activities may be therapeutically useful as a novel means of controlling breast cancer growth and invasiveness.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical Sciences, University of Ulsan College of Medicine, Seoul, 138-736, Republic of Korea.

ABSTRACT
Matrix metalloproteinase-9 (MMP-9) plays a central role in the invasion and metastasis of various types of cancer cells. Here, we demonstrate that glaucine, an alkaloid isolated from the plant Corydalis turtschaninovii tuber (Papaveraceae), can inhibit the migration and invasion of human breast cancer cells. We further show that glaucine significantly blocks phorbol 12-myristate 13-acetate (PMA)-induced MMP-9 expression and activity in a dose-dependent manner. Results from reporter gene and electrophoretic mobility shift assays revealed that glaucine inhibits MMP-9 expression by suppressing activation of the nuclear transcription factor nuclear factor-κB (NF-κB). Moreover, glaucine attenuates PMA-induced IκBα degradation and nuclear translocation of NF-κB. Finally, we also found that glaucine inhibits invasion and MMP-9 expression in the highly metastatic MDA-MB-231 breast cancer cell line. Taken together, our findings indicate that the MMP-9 inhibitory activity of glaucine and its abilities to attenuate IκBα and NF-κB activities may be therapeutically useful as a novel means of controlling breast cancer growth and invasiveness.

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Glaucine inhibits the NF-κB activity in PMA-stimulated MCF-7 cells. a MCF-7 cells were pretreated with the indicated concentration of glaucine for 30 min and incubated with 100 nM PMA for 30 min. Nuclear extracts were prepared and incubated with radiolabeled oligonucleotides containing the NF-κB motif in the MMP-9 promoter. b MCF-7 cells were pretreated with the indicated concentration of glaucine for 30 min and incubated with 100-nM PMA for 30 min. Cells were harvested and fractionated into the cytoplasm and the nucleus. Lysates were then separated on a 10 % SDS–polyacrylamide gel and subjected to western blotting with anti-p65, anti-p-IκBα, and anti-IκBα antibodies. The analysis was repeated in three times, and α-tubulin and PCNA were used as markers for the cytoplasmic and nuclear fractions. The NF-κB protein level that was translocated to the nucleus was quantified by densitometric analyses. The bars represent the mean ± SD. One-way ANOVA was performed to determine statistical significance (*P < 0.05)
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Fig5: Glaucine inhibits the NF-κB activity in PMA-stimulated MCF-7 cells. a MCF-7 cells were pretreated with the indicated concentration of glaucine for 30 min and incubated with 100 nM PMA for 30 min. Nuclear extracts were prepared and incubated with radiolabeled oligonucleotides containing the NF-κB motif in the MMP-9 promoter. b MCF-7 cells were pretreated with the indicated concentration of glaucine for 30 min and incubated with 100-nM PMA for 30 min. Cells were harvested and fractionated into the cytoplasm and the nucleus. Lysates were then separated on a 10 % SDS–polyacrylamide gel and subjected to western blotting with anti-p65, anti-p-IκBα, and anti-IκBα antibodies. The analysis was repeated in three times, and α-tubulin and PCNA were used as markers for the cytoplasmic and nuclear fractions. The NF-κB protein level that was translocated to the nucleus was quantified by densitometric analyses. The bars represent the mean ± SD. One-way ANOVA was performed to determine statistical significance (*P < 0.05)

Mentions: We next performed EMSAs to determine the effect of glaucine on the DNA-binding activity of NF-κB. As shown in Fig. 5a, PMA induced NF-κB DNA-binding activity, which was dose-dependently reduced by glaucine. To confirm this observation, we analyzed p65 levels in the cytosolic and nuclear fractions. The results showed that PMA increased IκBα phosphorylation in the cytoplasm, leading to increased nuclear translocation of the NF-κB p65 subunit (Fig. 5b). However, glaucine inhibited PMA-induced phosphorylation of IκBα in a dose-dependent manner (Fig. 5b) and decreased nuclear NF-κB p65 subunit levels (Fig. 5b, right panel). These results indicate that glaucine inhibits NF-κB activation by suppressing IκBα phosphorylation and the subsequent nuclear translocation of NF-κB in PMA-treated MCF-7 cells.Fig. 5


Glaucine inhibits breast cancer cell migration and invasion by inhibiting MMP-9 gene expression through the suppression of NF-κB activation.

Kang H, Jang SW, Pak JH, Shim S - Mol. Cell. Biochem. (2015)

Glaucine inhibits the NF-κB activity in PMA-stimulated MCF-7 cells. a MCF-7 cells were pretreated with the indicated concentration of glaucine for 30 min and incubated with 100 nM PMA for 30 min. Nuclear extracts were prepared and incubated with radiolabeled oligonucleotides containing the NF-κB motif in the MMP-9 promoter. b MCF-7 cells were pretreated with the indicated concentration of glaucine for 30 min and incubated with 100-nM PMA for 30 min. Cells were harvested and fractionated into the cytoplasm and the nucleus. Lysates were then separated on a 10 % SDS–polyacrylamide gel and subjected to western blotting with anti-p65, anti-p-IκBα, and anti-IκBα antibodies. The analysis was repeated in three times, and α-tubulin and PCNA were used as markers for the cytoplasmic and nuclear fractions. The NF-κB protein level that was translocated to the nucleus was quantified by densitometric analyses. The bars represent the mean ± SD. One-way ANOVA was performed to determine statistical significance (*P < 0.05)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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Fig5: Glaucine inhibits the NF-κB activity in PMA-stimulated MCF-7 cells. a MCF-7 cells were pretreated with the indicated concentration of glaucine for 30 min and incubated with 100 nM PMA for 30 min. Nuclear extracts were prepared and incubated with radiolabeled oligonucleotides containing the NF-κB motif in the MMP-9 promoter. b MCF-7 cells were pretreated with the indicated concentration of glaucine for 30 min and incubated with 100-nM PMA for 30 min. Cells were harvested and fractionated into the cytoplasm and the nucleus. Lysates were then separated on a 10 % SDS–polyacrylamide gel and subjected to western blotting with anti-p65, anti-p-IκBα, and anti-IκBα antibodies. The analysis was repeated in three times, and α-tubulin and PCNA were used as markers for the cytoplasmic and nuclear fractions. The NF-κB protein level that was translocated to the nucleus was quantified by densitometric analyses. The bars represent the mean ± SD. One-way ANOVA was performed to determine statistical significance (*P < 0.05)
Mentions: We next performed EMSAs to determine the effect of glaucine on the DNA-binding activity of NF-κB. As shown in Fig. 5a, PMA induced NF-κB DNA-binding activity, which was dose-dependently reduced by glaucine. To confirm this observation, we analyzed p65 levels in the cytosolic and nuclear fractions. The results showed that PMA increased IκBα phosphorylation in the cytoplasm, leading to increased nuclear translocation of the NF-κB p65 subunit (Fig. 5b). However, glaucine inhibited PMA-induced phosphorylation of IκBα in a dose-dependent manner (Fig. 5b) and decreased nuclear NF-κB p65 subunit levels (Fig. 5b, right panel). These results indicate that glaucine inhibits NF-κB activation by suppressing IκBα phosphorylation and the subsequent nuclear translocation of NF-κB in PMA-treated MCF-7 cells.Fig. 5

Bottom Line: We further show that glaucine significantly blocks phorbol 12-myristate 13-acetate (PMA)-induced MMP-9 expression and activity in a dose-dependent manner.Moreover, glaucine attenuates PMA-induced IκBα degradation and nuclear translocation of NF-κB.Taken together, our findings indicate that the MMP-9 inhibitory activity of glaucine and its abilities to attenuate IκBα and NF-κB activities may be therapeutically useful as a novel means of controlling breast cancer growth and invasiveness.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical Sciences, University of Ulsan College of Medicine, Seoul, 138-736, Republic of Korea.

ABSTRACT
Matrix metalloproteinase-9 (MMP-9) plays a central role in the invasion and metastasis of various types of cancer cells. Here, we demonstrate that glaucine, an alkaloid isolated from the plant Corydalis turtschaninovii tuber (Papaveraceae), can inhibit the migration and invasion of human breast cancer cells. We further show that glaucine significantly blocks phorbol 12-myristate 13-acetate (PMA)-induced MMP-9 expression and activity in a dose-dependent manner. Results from reporter gene and electrophoretic mobility shift assays revealed that glaucine inhibits MMP-9 expression by suppressing activation of the nuclear transcription factor nuclear factor-κB (NF-κB). Moreover, glaucine attenuates PMA-induced IκBα degradation and nuclear translocation of NF-κB. Finally, we also found that glaucine inhibits invasion and MMP-9 expression in the highly metastatic MDA-MB-231 breast cancer cell line. Taken together, our findings indicate that the MMP-9 inhibitory activity of glaucine and its abilities to attenuate IκBα and NF-κB activities may be therapeutically useful as a novel means of controlling breast cancer growth and invasiveness.

Show MeSH
Related in: MedlinePlus