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Ciprofloxacin Improves the Stemness of Human Dermal Papilla Cells.

Kiratipaiboon C, Tengamnuay P, Chanvorachote P - Stem Cells Int (2015)

Bottom Line: We found that ciprofloxacin exerted its effect through ATP-dependent tyrosine kinase/glycogen synthase kinase3β dependent mechanism which in turn upregulated β-catenin.The effects of ciprofloxacin in preserving stem cell features were confirmed in the primary dermal papilla cells directly obtained from human hair follicles.Together, these results revealed a novel application of ciprofloxacin for stem cell maintenance and provided the underlying mechanisms that are responsible for the stemness in dermal papilla cells.

View Article: PubMed Central - PubMed

Affiliation: Pharmaceutical Technology (International) Program, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand.

ABSTRACT
Improvement in the expansion method of adult stem cells may augment their use in regenerative therapy. Using human dermal papilla cell line as well as primary dermal papilla cells as model systems, the present study demonstrated that ciprofloxacin treatment could prevent the loss of stemness during culture. Clonogenicity and stem cell markers of dermal papilla cells were shown to gradually decrease in the culture in a time-dependent manner. Treatment of the cells with nontoxic concentrations of ciprofloxacin could maintain both stem cell morphology and clonogenicity, as well as all stem cells markers. We found that ciprofloxacin exerted its effect through ATP-dependent tyrosine kinase/glycogen synthase kinase3β dependent mechanism which in turn upregulated β-catenin. Besides, ciprofloxacin was shown to induce epithelial-mesenchymal transition in DPCs as the transcription factors ZEB1 and Snail were significantly increased. Furthermore, the self-renewal proteins of Wnt/β-catenin pathway, namely, Nanog and Oct-4 were significantly upregulated in the ciprofloxacin-treated cells. The effects of ciprofloxacin in preserving stem cell features were confirmed in the primary dermal papilla cells directly obtained from human hair follicles. Together, these results revealed a novel application of ciprofloxacin for stem cell maintenance and provided the underlying mechanisms that are responsible for the stemness in dermal papilla cells.

No MeSH data available.


Related in: MedlinePlus

Effects of CIP on Wnt/β-catenin signaling, EMT, and self-renewal transcription factors in DPCs. (a) Cells were treated with CIP (1–10 μg/mL) for 72 h. After indicated treatment, the levels of Wnt/β-catenin signaling (Akt, p-Akt (Ser 473), GSK3β, p-GSK3β (Ser 9), and β-catenin) were analyzed by western blot analysis. The immunoblot signals were quantified by densitometry and mean data from independent experiments were normalized to the results. (b) EMT and self-renewal transcription factors (ZEB1, Oct-4, Nanog, Slug, and Snail) were determined by western blot analysis. (c) Downstream targets of Snail including N-cadherin, vimentin, total Erk, and p-Erk (Thr 202/Tyr 204) were analyzed by western blot analysis. β-actin was used as the loading control. The western blot signals were quantified by densitometry and mean data from independent experiments were normalized to the results. The data represent the means of four independent samples ± SD. ∗P < 0.05 versus untreated control.
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fig4: Effects of CIP on Wnt/β-catenin signaling, EMT, and self-renewal transcription factors in DPCs. (a) Cells were treated with CIP (1–10 μg/mL) for 72 h. After indicated treatment, the levels of Wnt/β-catenin signaling (Akt, p-Akt (Ser 473), GSK3β, p-GSK3β (Ser 9), and β-catenin) were analyzed by western blot analysis. The immunoblot signals were quantified by densitometry and mean data from independent experiments were normalized to the results. (b) EMT and self-renewal transcription factors (ZEB1, Oct-4, Nanog, Slug, and Snail) were determined by western blot analysis. (c) Downstream targets of Snail including N-cadherin, vimentin, total Erk, and p-Erk (Thr 202/Tyr 204) were analyzed by western blot analysis. β-actin was used as the loading control. The western blot signals were quantified by densitometry and mean data from independent experiments were normalized to the results. The data represent the means of four independent samples ± SD. ∗P < 0.05 versus untreated control.

Mentions: The results showed that activated Akt was significantly enhanced by treatment of the cells with CIP at 2.5–10 μg/mL in a dose-dependent manner (Figure 4(a)). Consequently, the downstream GSK3β was inactivated by the treatment of CIP as indicated by an increase in phosphorylated GSK3β (Figure 4(a)). As GSK3β was shown to inhibit the degradation process of β-catenin, we found corresponding results that phosphorylated GSK3β leads to an increase of cellular β-catenin in CIP treated DPCs (Figure 4(a)). These data suggested that CIP maintains stemness in DPCs at least in part by increased cellular β-catenin via Akt/GSK3β pathway.


Ciprofloxacin Improves the Stemness of Human Dermal Papilla Cells.

Kiratipaiboon C, Tengamnuay P, Chanvorachote P - Stem Cells Int (2015)

Effects of CIP on Wnt/β-catenin signaling, EMT, and self-renewal transcription factors in DPCs. (a) Cells were treated with CIP (1–10 μg/mL) for 72 h. After indicated treatment, the levels of Wnt/β-catenin signaling (Akt, p-Akt (Ser 473), GSK3β, p-GSK3β (Ser 9), and β-catenin) were analyzed by western blot analysis. The immunoblot signals were quantified by densitometry and mean data from independent experiments were normalized to the results. (b) EMT and self-renewal transcription factors (ZEB1, Oct-4, Nanog, Slug, and Snail) were determined by western blot analysis. (c) Downstream targets of Snail including N-cadherin, vimentin, total Erk, and p-Erk (Thr 202/Tyr 204) were analyzed by western blot analysis. β-actin was used as the loading control. The western blot signals were quantified by densitometry and mean data from independent experiments were normalized to the results. The data represent the means of four independent samples ± SD. ∗P < 0.05 versus untreated control.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4663358&req=5

fig4: Effects of CIP on Wnt/β-catenin signaling, EMT, and self-renewal transcription factors in DPCs. (a) Cells were treated with CIP (1–10 μg/mL) for 72 h. After indicated treatment, the levels of Wnt/β-catenin signaling (Akt, p-Akt (Ser 473), GSK3β, p-GSK3β (Ser 9), and β-catenin) were analyzed by western blot analysis. The immunoblot signals were quantified by densitometry and mean data from independent experiments were normalized to the results. (b) EMT and self-renewal transcription factors (ZEB1, Oct-4, Nanog, Slug, and Snail) were determined by western blot analysis. (c) Downstream targets of Snail including N-cadherin, vimentin, total Erk, and p-Erk (Thr 202/Tyr 204) were analyzed by western blot analysis. β-actin was used as the loading control. The western blot signals were quantified by densitometry and mean data from independent experiments were normalized to the results. The data represent the means of four independent samples ± SD. ∗P < 0.05 versus untreated control.
Mentions: The results showed that activated Akt was significantly enhanced by treatment of the cells with CIP at 2.5–10 μg/mL in a dose-dependent manner (Figure 4(a)). Consequently, the downstream GSK3β was inactivated by the treatment of CIP as indicated by an increase in phosphorylated GSK3β (Figure 4(a)). As GSK3β was shown to inhibit the degradation process of β-catenin, we found corresponding results that phosphorylated GSK3β leads to an increase of cellular β-catenin in CIP treated DPCs (Figure 4(a)). These data suggested that CIP maintains stemness in DPCs at least in part by increased cellular β-catenin via Akt/GSK3β pathway.

Bottom Line: We found that ciprofloxacin exerted its effect through ATP-dependent tyrosine kinase/glycogen synthase kinase3β dependent mechanism which in turn upregulated β-catenin.The effects of ciprofloxacin in preserving stem cell features were confirmed in the primary dermal papilla cells directly obtained from human hair follicles.Together, these results revealed a novel application of ciprofloxacin for stem cell maintenance and provided the underlying mechanisms that are responsible for the stemness in dermal papilla cells.

View Article: PubMed Central - PubMed

Affiliation: Pharmaceutical Technology (International) Program, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand.

ABSTRACT
Improvement in the expansion method of adult stem cells may augment their use in regenerative therapy. Using human dermal papilla cell line as well as primary dermal papilla cells as model systems, the present study demonstrated that ciprofloxacin treatment could prevent the loss of stemness during culture. Clonogenicity and stem cell markers of dermal papilla cells were shown to gradually decrease in the culture in a time-dependent manner. Treatment of the cells with nontoxic concentrations of ciprofloxacin could maintain both stem cell morphology and clonogenicity, as well as all stem cells markers. We found that ciprofloxacin exerted its effect through ATP-dependent tyrosine kinase/glycogen synthase kinase3β dependent mechanism which in turn upregulated β-catenin. Besides, ciprofloxacin was shown to induce epithelial-mesenchymal transition in DPCs as the transcription factors ZEB1 and Snail were significantly increased. Furthermore, the self-renewal proteins of Wnt/β-catenin pathway, namely, Nanog and Oct-4 were significantly upregulated in the ciprofloxacin-treated cells. The effects of ciprofloxacin in preserving stem cell features were confirmed in the primary dermal papilla cells directly obtained from human hair follicles. Together, these results revealed a novel application of ciprofloxacin for stem cell maintenance and provided the underlying mechanisms that are responsible for the stemness in dermal papilla cells.

No MeSH data available.


Related in: MedlinePlus