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Triclosan potentiates epithelial-to-mesenchymal transition in anoikis-resistant human lung cancer cells.

Winitthana T, Lawanprasert S, Chanvorachote P - PLoS ONE (2014)

Bottom Line: Herein, we report the effect of triclosan, a widely used antibacterial agent found in many daily products, in enhancing the epithelial-to-mesenchymal transition (EMT) in aggressive anoikis resistant human H460 lung cancer cells.Also, the mesenchymal-like morphology and decrease in cell-to-cell adhesion were observed in triclosan-treated cells.Importantly, western blot analysis revealed that triclosan-treated cells exhibited decreased E-cadherin, while the levels of EMT markers, namely N-cadherin, vimentin, snail and slug were found to be significantly up-regulated.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology and Physiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand.

ABSTRACT
Alteration of cancer cell toward mesenchymal phenotype has been shown to potentiate tumor aggressiveness by increasing cancer cell metastasis. Herein, we report the effect of triclosan, a widely used antibacterial agent found in many daily products, in enhancing the epithelial-to-mesenchymal transition (EMT) in aggressive anoikis resistant human H460 lung cancer cells. EMT has been long known to increase abilities of the cells to increase migration, invasion, and survival in circulating system. The present study reveals that treatment of the cancer cells with triclosan at the physiologically related concentrations significantly increased the colony number of the cancer cells assessed by tumor formation assay. Also, the mesenchymal-like morphology and decrease in cell-to-cell adhesion were observed in triclosan-treated cells. Importantly, western blot analysis revealed that triclosan-treated cells exhibited decreased E-cadherin, while the levels of EMT markers, namely N-cadherin, vimentin, snail and slug were found to be significantly up-regulated. Furthermore, EMT induced by triclosan treatment was accompanied by the activation of focal adhesion kinase/ATP dependent tyrosine kinase (FAK/Akt) and Ras-related C3 botulinum toxin substrate 1 (Rac1), which enhanced the ability of the cells to migrate and invade. In conclusion, we demonstrated for the first time that triclosan may potentiate cancer cells survival in detached condition and motility via the process of EMT. As mentioned capabilities are required for success in metastasis, the present study provides the novel toxicological information and encourages the awareness of triclosan use in cancer patients.

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Related in: MedlinePlus

Cytotoxic effect and proliferative effect of TCS on anoikis resistant H460 cells.(A) The chemical structure of TCS. (B) Multicellular aggregation of anoikis resistant H460 cells. Scale bar is 1,000 µm. (C–D) After treatment with TCS (0–10 µM) for 24 h, the percentage of cell viability was determined by MTT assay and the percentage of apoptotic cells was detected by Hoechst33342 staining, respectively. Values are means of the three independent triplicate samples ± SE. *P<0.05 versus non-treated control. (E) After the indicated treatment, nuclear morphology of the cells was detected by Hoechst33342/PI co-staining assay and visualized under a fluorescence microscope. Scale bar is 50 µm. (F) Cells were treated with TCS (0–7.5 µM) for 24, 48 or 72 h. Cell viability was determined by MTT assay. The data represent the means of the three independent triplicate samples ± SE. *P<0.05 versus non-treated control at each indicated time. (G–H) Cells were treated with TCS (0–7.5 µM) for 48 h. Cell cycle of TCS-treated cells was determined by PI staining and flow cytometry. *P<0.05 versus non-treated control.
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pone-0110851-g001: Cytotoxic effect and proliferative effect of TCS on anoikis resistant H460 cells.(A) The chemical structure of TCS. (B) Multicellular aggregation of anoikis resistant H460 cells. Scale bar is 1,000 µm. (C–D) After treatment with TCS (0–10 µM) for 24 h, the percentage of cell viability was determined by MTT assay and the percentage of apoptotic cells was detected by Hoechst33342 staining, respectively. Values are means of the three independent triplicate samples ± SE. *P<0.05 versus non-treated control. (E) After the indicated treatment, nuclear morphology of the cells was detected by Hoechst33342/PI co-staining assay and visualized under a fluorescence microscope. Scale bar is 50 µm. (F) Cells were treated with TCS (0–7.5 µM) for 24, 48 or 72 h. Cell viability was determined by MTT assay. The data represent the means of the three independent triplicate samples ± SE. *P<0.05 versus non-treated control at each indicated time. (G–H) Cells were treated with TCS (0–7.5 µM) for 48 h. Cell cycle of TCS-treated cells was determined by PI staining and flow cytometry. *P<0.05 versus non-treated control.

Mentions: The well-known broad-spectrum anti-bacterial agent triclosan (2,4,4′ –trichloro-2′-hydroxydiphenyl ether; TCS) (Figure 1A) has been commercially used in a variety of products to inhibit the growth of bacteria, fungi, and mildew [1], [2]. TCS has been used under the regulation of the Food and Drug Administration (in cosmetics, deodorant, hand soaps, toothpaste) as well as the Environmental Protection Agency (in materials preservative incorporated into household plastics and textiles) [2], [3]. The concentrations used of TCS in different products may vary; however, its levels in most personal care products range from 0.1–2% [1], [3]. The fact that the significant levels of TCS are detectable in the plasma of TCS-exposed human at the concentration ranging from 0.02 and 20 µg/ml (0.069 and 69 µM) leads to the possible conception that this agent may possibly impact human physiology [4].


Triclosan potentiates epithelial-to-mesenchymal transition in anoikis-resistant human lung cancer cells.

Winitthana T, Lawanprasert S, Chanvorachote P - PLoS ONE (2014)

Cytotoxic effect and proliferative effect of TCS on anoikis resistant H460 cells.(A) The chemical structure of TCS. (B) Multicellular aggregation of anoikis resistant H460 cells. Scale bar is 1,000 µm. (C–D) After treatment with TCS (0–10 µM) for 24 h, the percentage of cell viability was determined by MTT assay and the percentage of apoptotic cells was detected by Hoechst33342 staining, respectively. Values are means of the three independent triplicate samples ± SE. *P<0.05 versus non-treated control. (E) After the indicated treatment, nuclear morphology of the cells was detected by Hoechst33342/PI co-staining assay and visualized under a fluorescence microscope. Scale bar is 50 µm. (F) Cells were treated with TCS (0–7.5 µM) for 24, 48 or 72 h. Cell viability was determined by MTT assay. The data represent the means of the three independent triplicate samples ± SE. *P<0.05 versus non-treated control at each indicated time. (G–H) Cells were treated with TCS (0–7.5 µM) for 48 h. Cell cycle of TCS-treated cells was determined by PI staining and flow cytometry. *P<0.05 versus non-treated control.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4199721&req=5

pone-0110851-g001: Cytotoxic effect and proliferative effect of TCS on anoikis resistant H460 cells.(A) The chemical structure of TCS. (B) Multicellular aggregation of anoikis resistant H460 cells. Scale bar is 1,000 µm. (C–D) After treatment with TCS (0–10 µM) for 24 h, the percentage of cell viability was determined by MTT assay and the percentage of apoptotic cells was detected by Hoechst33342 staining, respectively. Values are means of the three independent triplicate samples ± SE. *P<0.05 versus non-treated control. (E) After the indicated treatment, nuclear morphology of the cells was detected by Hoechst33342/PI co-staining assay and visualized under a fluorescence microscope. Scale bar is 50 µm. (F) Cells were treated with TCS (0–7.5 µM) for 24, 48 or 72 h. Cell viability was determined by MTT assay. The data represent the means of the three independent triplicate samples ± SE. *P<0.05 versus non-treated control at each indicated time. (G–H) Cells were treated with TCS (0–7.5 µM) for 48 h. Cell cycle of TCS-treated cells was determined by PI staining and flow cytometry. *P<0.05 versus non-treated control.
Mentions: The well-known broad-spectrum anti-bacterial agent triclosan (2,4,4′ –trichloro-2′-hydroxydiphenyl ether; TCS) (Figure 1A) has been commercially used in a variety of products to inhibit the growth of bacteria, fungi, and mildew [1], [2]. TCS has been used under the regulation of the Food and Drug Administration (in cosmetics, deodorant, hand soaps, toothpaste) as well as the Environmental Protection Agency (in materials preservative incorporated into household plastics and textiles) [2], [3]. The concentrations used of TCS in different products may vary; however, its levels in most personal care products range from 0.1–2% [1], [3]. The fact that the significant levels of TCS are detectable in the plasma of TCS-exposed human at the concentration ranging from 0.02 and 20 µg/ml (0.069 and 69 µM) leads to the possible conception that this agent may possibly impact human physiology [4].

Bottom Line: Herein, we report the effect of triclosan, a widely used antibacterial agent found in many daily products, in enhancing the epithelial-to-mesenchymal transition (EMT) in aggressive anoikis resistant human H460 lung cancer cells.Also, the mesenchymal-like morphology and decrease in cell-to-cell adhesion were observed in triclosan-treated cells.Importantly, western blot analysis revealed that triclosan-treated cells exhibited decreased E-cadherin, while the levels of EMT markers, namely N-cadherin, vimentin, snail and slug were found to be significantly up-regulated.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology and Physiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand.

ABSTRACT
Alteration of cancer cell toward mesenchymal phenotype has been shown to potentiate tumor aggressiveness by increasing cancer cell metastasis. Herein, we report the effect of triclosan, a widely used antibacterial agent found in many daily products, in enhancing the epithelial-to-mesenchymal transition (EMT) in aggressive anoikis resistant human H460 lung cancer cells. EMT has been long known to increase abilities of the cells to increase migration, invasion, and survival in circulating system. The present study reveals that treatment of the cancer cells with triclosan at the physiologically related concentrations significantly increased the colony number of the cancer cells assessed by tumor formation assay. Also, the mesenchymal-like morphology and decrease in cell-to-cell adhesion were observed in triclosan-treated cells. Importantly, western blot analysis revealed that triclosan-treated cells exhibited decreased E-cadherin, while the levels of EMT markers, namely N-cadherin, vimentin, snail and slug were found to be significantly up-regulated. Furthermore, EMT induced by triclosan treatment was accompanied by the activation of focal adhesion kinase/ATP dependent tyrosine kinase (FAK/Akt) and Ras-related C3 botulinum toxin substrate 1 (Rac1), which enhanced the ability of the cells to migrate and invade. In conclusion, we demonstrated for the first time that triclosan may potentiate cancer cells survival in detached condition and motility via the process of EMT. As mentioned capabilities are required for success in metastasis, the present study provides the novel toxicological information and encourages the awareness of triclosan use in cancer patients.

Show MeSH
Related in: MedlinePlus