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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 activity of LfcinB(20–25)4 in long-term treatments. (a) SCC15 and CAL27 cells were incubated with the peptide for 24, 48, 72, and 96 h. (b) SCC15 and CAL27 cells were treated for 24 h with the peptide and washed, and cells were incubated for 0, 24, 48, and 72 h in fresh culture medium. After the treatments, cell viability was determined by the MTT assay and calculated as the percentage of average absorbance of each treatment relative to the average absorbance of the negative control. Cell viability was evaluated 24, 48, and 72 h after treatments and was calculated as the percentage of average absorbance of treatments in relation to average absorbance of negative control. The concentrations of LfcinB(20–25)4 and STA used were 22.25 μM (100 μg/mL) and 0.86 μM (0.4 μg/mL), respectively. Treatments were done in triplicate.
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fig3: Cytotoxic activity of LfcinB(20–25)4 in long-term treatments. (a) SCC15 and CAL27 cells were incubated with the peptide for 24, 48, 72, and 96 h. (b) SCC15 and CAL27 cells were treated for 24 h with the peptide and washed, and cells were incubated for 0, 24, 48, and 72 h in fresh culture medium. After the treatments, cell viability was determined by the MTT assay and calculated as the percentage of average absorbance of each treatment relative to the average absorbance of the negative control. Cell viability was evaluated 24, 48, and 72 h after treatments and was calculated as the percentage of average absorbance of treatments in relation to average absorbance of negative control. The concentrations of LfcinB(20–25)4 and STA used were 22.25 μM (100 μg/mL) and 0.86 μM (0.4 μg/mL), respectively. Treatments were done in triplicate.

Mentions: As has been previously observed with other therapeutic compounds, not all cells died after the various peptide treatments. The reasons for this are unknown and certainly varied. In order to explore this issue, we determined the proliferation capacity of the remaining cells after longer treatment periods with LfcinB(20–25)4 (Figure 3). The cytotoxic effect of LfcinB(20–25)4 in CAL27 was 98% after 24 h of incubation and progressively declined to 85% (after 72 h) and 62% (after 96 h) (Figure 3(a)). This progressive reduction was not observed in SCC15 cells, showing a rapid (from 24 h to 48 h period) recovery of viability (near 64%) which is maintained hereinafter; the reasons for this difference are unknown, but since SCC15 cells have a slower proliferation rate (42 h for SCC15 compared to 28 h for CAL27), this could explain in part the lack of progressive recovery; also the SSC15 cell line seems to have a higher resistance to STA treatment (Figure 3(a)). These results suggest that LfcinB(20–25)4 could exert its maximum effect before 24 h of treatment. In order to further investigate this, OSCC cells were treated for 24 h with the peptide as described previously, washed, and then cultured with fresh medium without peptides (Figure 3(b)). These results were very similar to those found in long-term treatments, confirming that its effect is produced before 24 h of treatment. In fact, LfcinB(20–25)4 exhibited a significant cytotoxic effect from the first hour of treatment in CAL27 (80%) and SCC15 (95%) (Figures 4(a) and 4(b)). A lower effect (42%) was found in the Het-1A cell line (Figure 4(c)) after 1 h of treatment with LfcinB(20–25)4 and this could not be differentiated from the STA treatment (Figure 4(c)); this suggests some specificity of the tetrameric peptide towards the tumorigenic cell lines.


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 activity of LfcinB(20–25)4 in long-term treatments. (a) SCC15 and CAL27 cells were incubated with the peptide for 24, 48, 72, and 96 h. (b) SCC15 and CAL27 cells were treated for 24 h with the peptide and washed, and cells were incubated for 0, 24, 48, and 72 h in fresh culture medium. After the treatments, cell viability was determined by the MTT assay and calculated as the percentage of average absorbance of each treatment relative to the average absorbance of the negative control. Cell viability was evaluated 24, 48, and 72 h after treatments and was calculated as the percentage of average absorbance of treatments in relation to average absorbance of negative control. The concentrations of LfcinB(20–25)4 and STA used were 22.25 μM (100 μg/mL) and 0.86 μM (0.4 μg/mL), respectively. Treatments were done in triplicate.
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fig3: Cytotoxic activity of LfcinB(20–25)4 in long-term treatments. (a) SCC15 and CAL27 cells were incubated with the peptide for 24, 48, 72, and 96 h. (b) SCC15 and CAL27 cells were treated for 24 h with the peptide and washed, and cells were incubated for 0, 24, 48, and 72 h in fresh culture medium. After the treatments, cell viability was determined by the MTT assay and calculated as the percentage of average absorbance of each treatment relative to the average absorbance of the negative control. Cell viability was evaluated 24, 48, and 72 h after treatments and was calculated as the percentage of average absorbance of treatments in relation to average absorbance of negative control. The concentrations of LfcinB(20–25)4 and STA used were 22.25 μM (100 μg/mL) and 0.86 μM (0.4 μg/mL), respectively. Treatments were done in triplicate.
Mentions: As has been previously observed with other therapeutic compounds, not all cells died after the various peptide treatments. The reasons for this are unknown and certainly varied. In order to explore this issue, we determined the proliferation capacity of the remaining cells after longer treatment periods with LfcinB(20–25)4 (Figure 3). The cytotoxic effect of LfcinB(20–25)4 in CAL27 was 98% after 24 h of incubation and progressively declined to 85% (after 72 h) and 62% (after 96 h) (Figure 3(a)). This progressive reduction was not observed in SCC15 cells, showing a rapid (from 24 h to 48 h period) recovery of viability (near 64%) which is maintained hereinafter; the reasons for this difference are unknown, but since SCC15 cells have a slower proliferation rate (42 h for SCC15 compared to 28 h for CAL27), this could explain in part the lack of progressive recovery; also the SSC15 cell line seems to have a higher resistance to STA treatment (Figure 3(a)). These results suggest that LfcinB(20–25)4 could exert its maximum effect before 24 h of treatment. In order to further investigate this, OSCC cells were treated for 24 h with the peptide as described previously, washed, and then cultured with fresh medium without peptides (Figure 3(b)). These results were very similar to those found in long-term treatments, confirming that its effect is produced before 24 h of treatment. In fact, LfcinB(20–25)4 exhibited a significant cytotoxic effect from the first hour of treatment in CAL27 (80%) and SCC15 (95%) (Figures 4(a) and 4(b)). A lower effect (42%) was found in the Het-1A cell line (Figure 4(c)) after 1 h of treatment with LfcinB(20–25)4 and this could not be differentiated from the STA treatment (Figure 4(c)); this suggests some specificity of the tetrameric peptide towards the tumorigenic cell lines.

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