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Biotransformed soybean extract (BSE) inhibits melanoma cell growth and viability in vitro: involvement of nuclear factor-kappa B signaling.

Vilela FM, Syed DN, Chamcheu JC, Calvo-Castro LA, Fortes VS, Fonseca MJ, Mukhtar H - PLoS ONE (2014)

Bottom Line: Further studies in A375 cells showed that decrease in cell viability with BSE treatment (1.5-1.9 mg/ml; 24 h) was associated with induction of apoptosis.Immunoblot analysis revealed that BSE treatment resulted in induction of PARP cleavage, activation of caspase-3, -7, and -8 and increased expression of TRAIL and its receptor DR4.BSE did not activate the intrinsic apoptotic pathway in A375 cells, as no change was observed in caspase-9 expression.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, Brazil; Department of Dermatology, University of Wisconsin, Madison, Wisconsin, United States of America.

ABSTRACT
Melanoma is recognized as one of the most aggressive cancers with a relatively high propensity for metastasis. The prognosis of melanoma remains poor in spite of treatment advances, emphasizing the importance of additional preventive measures. Isoflavonoids have become not only potential chemopreventive, but also important therapeutic natural agents. We evaluated the antiproliferative and proapoptotic properties of biotransformed soybean extract (BSE) in A375 melanoma cells. Previous analyses demonstrated that the concentration of daidzein, genistein and aminoacids/peptides present in BSE, fermented by Aspergillus awamori is much higher than in the non biotransformed extract (NBSE). Experiments comparing the efficacy of the extracts in preventing cancer cell growth showed that treatment (24 h) of aggressive melanoma cells (A375 and 451Lu) with BSE resulted in a dose-dependent inhibition of growth and viability. In contrast, treatment with similar doses of NBSE failed to inhibit melanoma cell viability. Further studies in A375 cells showed that decrease in cell viability with BSE treatment (1.5-1.9 mg/ml; 24 h) was associated with induction of apoptosis. Immunoblot analysis revealed that BSE treatment resulted in induction of PARP cleavage, activation of caspase-3, -7, and -8 and increased expression of TRAIL and its receptor DR4. BSE did not activate the intrinsic apoptotic pathway in A375 cells, as no change was observed in caspase-9 expression. The expression of Bcl-2 apoptotic proteins such as Bid and Bax remained unaffected with BSE treated cells. Interestingly, we also showed that BSE treatment increased the phosphorylation and activation of IKK, IκBα degradation and p65/NF-κB translocation to the nucleus, and that stimulation of the NF-???B pathway was required for BSE-induced apoptosis of A375 cells. Our findings indicate that the biotransformation of soybean plays a crucial role in the extract anti-cancer effect observed in melanoma cells. However, further studies are warranted to define the active anti-cancer agent(s) present in BSE.

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BSE and NBSE effect on melanoma cells growth and viability.451Lu and A375 cell lines were treated with NBSE and BSE for 24(DMSO)-treated cells were regarded as 100% viable. Data is represented as mean ± SE of 3 independent experiments. *p<0.05 versus NBSE.
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pone-0103248-g001: BSE and NBSE effect on melanoma cells growth and viability.451Lu and A375 cell lines were treated with NBSE and BSE for 24(DMSO)-treated cells were regarded as 100% viable. Data is represented as mean ± SE of 3 independent experiments. *p<0.05 versus NBSE.

Mentions: We first investigated the dose-dependent effect of BSE and NBSE treatment on the growth of 451Lu and A375 human melanoma cells using the MTT assay. BSE treatment of 451Lu (0–1.6 mg/mL) and A375 (0–2.2 mg/mL) melanoma cells for 24 h displayed a dose dependent decrease in cell growth/viability (Figure 1). The IC50 of BSE was estimated to be 0.8 mg/mL for 451LU and 1.7 mg/mL for A375 at 24 h, while NBSE treatment in both cell lines had no effect (in comparison to the untreated control) on cell viability at these doses. These data led to the selection of 1.5 to 1.9 mg/mL BSE doses for further mechanistic studies in the highly aggressive A375 melanoma cell line.


Biotransformed soybean extract (BSE) inhibits melanoma cell growth and viability in vitro: involvement of nuclear factor-kappa B signaling.

Vilela FM, Syed DN, Chamcheu JC, Calvo-Castro LA, Fortes VS, Fonseca MJ, Mukhtar H - PLoS ONE (2014)

BSE and NBSE effect on melanoma cells growth and viability.451Lu and A375 cell lines were treated with NBSE and BSE for 24(DMSO)-treated cells were regarded as 100% viable. Data is represented as mean ± SE of 3 independent experiments. *p<0.05 versus NBSE.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4114525&req=5

pone-0103248-g001: BSE and NBSE effect on melanoma cells growth and viability.451Lu and A375 cell lines were treated with NBSE and BSE for 24(DMSO)-treated cells were regarded as 100% viable. Data is represented as mean ± SE of 3 independent experiments. *p<0.05 versus NBSE.
Mentions: We first investigated the dose-dependent effect of BSE and NBSE treatment on the growth of 451Lu and A375 human melanoma cells using the MTT assay. BSE treatment of 451Lu (0–1.6 mg/mL) and A375 (0–2.2 mg/mL) melanoma cells for 24 h displayed a dose dependent decrease in cell growth/viability (Figure 1). The IC50 of BSE was estimated to be 0.8 mg/mL for 451LU and 1.7 mg/mL for A375 at 24 h, while NBSE treatment in both cell lines had no effect (in comparison to the untreated control) on cell viability at these doses. These data led to the selection of 1.5 to 1.9 mg/mL BSE doses for further mechanistic studies in the highly aggressive A375 melanoma cell line.

Bottom Line: Further studies in A375 cells showed that decrease in cell viability with BSE treatment (1.5-1.9 mg/ml; 24 h) was associated with induction of apoptosis.Immunoblot analysis revealed that BSE treatment resulted in induction of PARP cleavage, activation of caspase-3, -7, and -8 and increased expression of TRAIL and its receptor DR4.BSE did not activate the intrinsic apoptotic pathway in A375 cells, as no change was observed in caspase-9 expression.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, Brazil; Department of Dermatology, University of Wisconsin, Madison, Wisconsin, United States of America.

ABSTRACT
Melanoma is recognized as one of the most aggressive cancers with a relatively high propensity for metastasis. The prognosis of melanoma remains poor in spite of treatment advances, emphasizing the importance of additional preventive measures. Isoflavonoids have become not only potential chemopreventive, but also important therapeutic natural agents. We evaluated the antiproliferative and proapoptotic properties of biotransformed soybean extract (BSE) in A375 melanoma cells. Previous analyses demonstrated that the concentration of daidzein, genistein and aminoacids/peptides present in BSE, fermented by Aspergillus awamori is much higher than in the non biotransformed extract (NBSE). Experiments comparing the efficacy of the extracts in preventing cancer cell growth showed that treatment (24 h) of aggressive melanoma cells (A375 and 451Lu) with BSE resulted in a dose-dependent inhibition of growth and viability. In contrast, treatment with similar doses of NBSE failed to inhibit melanoma cell viability. Further studies in A375 cells showed that decrease in cell viability with BSE treatment (1.5-1.9 mg/ml; 24 h) was associated with induction of apoptosis. Immunoblot analysis revealed that BSE treatment resulted in induction of PARP cleavage, activation of caspase-3, -7, and -8 and increased expression of TRAIL and its receptor DR4. BSE did not activate the intrinsic apoptotic pathway in A375 cells, as no change was observed in caspase-9 expression. The expression of Bcl-2 apoptotic proteins such as Bid and Bax remained unaffected with BSE treated cells. Interestingly, we also showed that BSE treatment increased the phosphorylation and activation of IKK, IκBα degradation and p65/NF-κB translocation to the nucleus, and that stimulation of the NF-???B pathway was required for BSE-induced apoptosis of A375 cells. Our findings indicate that the biotransformation of soybean plays a crucial role in the extract anti-cancer effect observed in melanoma cells. However, further studies are warranted to define the active anti-cancer agent(s) present in BSE.

Show MeSH
Related in: MedlinePlus