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A Tetrameric Peptide Derived from Bovine Lactoferricin Exhibits Specific Cytotoxic Effects against Oral Squamous-Cell Carcinoma Cell Lines.

Solarte VA, Rosas JE, Rivera ZJ, Arango-Rodríguez ML, García JE, Vernot JP - Biomed Res Int (2015)

Bottom Line: Additionally, this tetrameric peptide showed the highest specificity towards tumorigenic cells among the tested peptides.Interestingly, this effect was very fast, with cell shrinkage, severe damage to cell membrane permeability, and lysis within one hour of treatment.Our results are consistent with a necrotic effect rather than an apoptotic one and suggest that this tetrameric peptide could be considered as a new candidate for the therapeutic treatment of OSCC.

View Article: PubMed Central - PubMed

Affiliation: Cellular and Molecular Physiology Group, Biomedical Research Institute, Faculty of Medicine, Universidad Nacional de Colombia, Bogotá 111321, Colombia.

ABSTRACT
Several short linear peptides derived from cyclic bovine lactoferricin were synthesized and tested for their cytotoxic effect against the oral cavity squamous-cell carcinoma (OSCC) cell lines CAL27 and SCC15. As a control, an immortalized and nontumorigenic cell line, Het-1A, was used. Linear peptides based on the RRWQWR core sequence showed a moderate cytotoxic effect and specificity towards tumorigenic cells. A tetrameric peptide, LfcinB(20-25)4, containing the RRWQWR motif, exhibited greater cytotoxic activity (>90%) in both OSCC cell lines compared to the linear lactoferricin peptide or the lactoferrin protein. Additionally, this tetrameric peptide showed the highest specificity towards tumorigenic cells among the tested peptides. Interestingly, this effect was very fast, with cell shrinkage, severe damage to cell membrane permeability, and lysis within one hour of treatment. Our results are consistent with a necrotic effect rather than an apoptotic one and suggest that this tetrameric peptide could be considered as a new candidate for the therapeutic treatment of OSCC.

No MeSH data available.


Related in: MedlinePlus

Cytotoxic effect of LfB and LfcinB25 in the OSCC tumorigenic cell lines CAL27 (a) and SCC15 (b) and the immortalized nontumorigenic keratinocytes cell line Het-1A (c). The cells were incubated for 24 h with the LfB protein and the LfcinB25 peptide. After treatment, cell viability was determined by MTT assay and calculated as the percentage of average absorbance of each treatment relative to the average absorbance of the negative control. The maximum concentration of the LfB protein was 1.25 μM (100 μg/mL) and of LfcinB25 32 μM (100 μg/mL). Each treatment was done in triplicate.
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fig1: Cytotoxic effect of LfB and LfcinB25 in the OSCC tumorigenic cell lines CAL27 (a) and SCC15 (b) and the immortalized nontumorigenic keratinocytes cell line Het-1A (c). The cells were incubated for 24 h with the LfB protein and the LfcinB25 peptide. After treatment, cell viability was determined by MTT assay and calculated as the percentage of average absorbance of each treatment relative to the average absorbance of the negative control. The maximum concentration of the LfB protein was 1.25 μM (100 μg/mL) and of LfcinB25 32 μM (100 μg/mL). Each treatment was done in triplicate.

Mentions: MTT cell viability assays showed that LfB was cytotoxic to CAL27 cells in a dose-dependent manner, showing 56.6% cytotoxicity at the highest concentration tested (1.25 μM equivalent to 100 μg/mL) (Figure 1(a), solid line). Only a 11.2% cytotoxicity was detected at the same concentration in the SCC15 cell line (Figure 1(b), solid line). In the nontumorigenic cell line Het-1A, the LfB cytotoxicity was 25.5% at the maximum concentration tested (Figure 1(c), solid line). Since it has been suggested that the LfB cytotoxic effect relies mainly on the N-terminal region of the protein, the LfcinB25 peptide was synthesized and its cytotoxic activity determined by the MTT assay. This N-terminal linear peptide exhibited a similar and dose-dependent cytotoxic effect both in OSCC cell lines (64.4% and 60% for CAL27 and SCC15, resp.) and in the nontumorigenic cell line Het-1A (60.5%) at the highest concentration tested (32 μM, equivalent to 100 μg/mL) (Figure 1, dotted line).


A Tetrameric Peptide Derived from Bovine Lactoferricin Exhibits Specific Cytotoxic Effects against Oral Squamous-Cell Carcinoma Cell Lines.

Solarte VA, Rosas JE, Rivera ZJ, Arango-Rodríguez ML, García JE, Vernot JP - Biomed Res Int (2015)

Cytotoxic effect of LfB and LfcinB25 in the OSCC tumorigenic cell lines CAL27 (a) and SCC15 (b) and the immortalized nontumorigenic keratinocytes cell line Het-1A (c). The cells were incubated for 24 h with the LfB protein and the LfcinB25 peptide. After treatment, cell viability was determined by MTT assay and calculated as the percentage of average absorbance of each treatment relative to the average absorbance of the negative control. The maximum concentration of the LfB protein was 1.25 μM (100 μg/mL) and of LfcinB25 32 μM (100 μg/mL). Each treatment was done in triplicate.
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Related In: Results  -  Collection

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fig1: Cytotoxic effect of LfB and LfcinB25 in the OSCC tumorigenic cell lines CAL27 (a) and SCC15 (b) and the immortalized nontumorigenic keratinocytes cell line Het-1A (c). The cells were incubated for 24 h with the LfB protein and the LfcinB25 peptide. After treatment, cell viability was determined by MTT assay and calculated as the percentage of average absorbance of each treatment relative to the average absorbance of the negative control. The maximum concentration of the LfB protein was 1.25 μM (100 μg/mL) and of LfcinB25 32 μM (100 μg/mL). Each treatment was done in triplicate.
Mentions: MTT cell viability assays showed that LfB was cytotoxic to CAL27 cells in a dose-dependent manner, showing 56.6% cytotoxicity at the highest concentration tested (1.25 μM equivalent to 100 μg/mL) (Figure 1(a), solid line). Only a 11.2% cytotoxicity was detected at the same concentration in the SCC15 cell line (Figure 1(b), solid line). In the nontumorigenic cell line Het-1A, the LfB cytotoxicity was 25.5% at the maximum concentration tested (Figure 1(c), solid line). Since it has been suggested that the LfB cytotoxic effect relies mainly on the N-terminal region of the protein, the LfcinB25 peptide was synthesized and its cytotoxic activity determined by the MTT assay. This N-terminal linear peptide exhibited a similar and dose-dependent cytotoxic effect both in OSCC cell lines (64.4% and 60% for CAL27 and SCC15, resp.) and in the nontumorigenic cell line Het-1A (60.5%) at the highest concentration tested (32 μM, equivalent to 100 μg/mL) (Figure 1, dotted line).

Bottom Line: Additionally, this tetrameric peptide showed the highest specificity towards tumorigenic cells among the tested peptides.Interestingly, this effect was very fast, with cell shrinkage, severe damage to cell membrane permeability, and lysis within one hour of treatment.Our results are consistent with a necrotic effect rather than an apoptotic one and suggest that this tetrameric peptide could be considered as a new candidate for the therapeutic treatment of OSCC.

View Article: PubMed Central - PubMed

Affiliation: Cellular and Molecular Physiology Group, Biomedical Research Institute, Faculty of Medicine, Universidad Nacional de Colombia, Bogotá 111321, Colombia.

ABSTRACT
Several short linear peptides derived from cyclic bovine lactoferricin were synthesized and tested for their cytotoxic effect against the oral cavity squamous-cell carcinoma (OSCC) cell lines CAL27 and SCC15. As a control, an immortalized and nontumorigenic cell line, Het-1A, was used. Linear peptides based on the RRWQWR core sequence showed a moderate cytotoxic effect and specificity towards tumorigenic cells. A tetrameric peptide, LfcinB(20-25)4, containing the RRWQWR motif, exhibited greater cytotoxic activity (>90%) in both OSCC cell lines compared to the linear lactoferricin peptide or the lactoferrin protein. Additionally, this tetrameric peptide showed the highest specificity towards tumorigenic cells among the tested peptides. Interestingly, this effect was very fast, with cell shrinkage, severe damage to cell membrane permeability, and lysis within one hour of treatment. Our results are consistent with a necrotic effect rather than an apoptotic one and suggest that this tetrameric peptide could be considered as a new candidate for the therapeutic treatment of OSCC.

No MeSH data available.


Related in: MedlinePlus