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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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Effects of TCS-mediated EMT on migratory-related proteins.(A) Anoikis resistant H460 cells were treated with TCS at 0–7.5 µM for 24 h in detached condition and then attached on conventional culture dishes for 4 h. The level of pFAK (Tyr 397), FAK, pAkt (Ser 473), Akt, activated Rac1 (Rac1-GTP) and activated RhoA (RhoA-GTP) were determined by western blotting. Blots were reprobed with β-actin to confirm equal loading. (B) The immunoblot signals were quantified by densitometry and mean data from independent experiments were normalized to the results. Values are means of the three triplicate independent samples ± SE. *P<0.05 versus non-treated control.
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pone-0110851-g006: Effects of TCS-mediated EMT on migratory-related proteins.(A) Anoikis resistant H460 cells were treated with TCS at 0–7.5 µM for 24 h in detached condition and then attached on conventional culture dishes for 4 h. The level of pFAK (Tyr 397), FAK, pAkt (Ser 473), Akt, activated Rac1 (Rac1-GTP) and activated RhoA (RhoA-GTP) were determined by western blotting. Blots were reprobed with β-actin to confirm equal loading. (B) The immunoblot signals were quantified by densitometry and mean data from independent experiments were normalized to the results. Values are means of the three triplicate independent samples ± SE. *P<0.05 versus non-treated control.

Mentions: Furthermore, the down-stream effector proteins which are responsible for cell motility were determined using western blotting. The cells were treated with indicated concentrations of TCS for 24 h and subjected to western blot analysis. The expression levels of migratory-related proteins including activated FAK (phosphorylated FAK, Tyr 397), FAK, activated Akt (phosphorylated Akt, Ser 473), Akt, activated Rac1 (Rac1-GTP), and activated RhoA (RhoA-GTP) were investigated. Figures 6A and B show that TCS treatment significantly increased the level of phosphorylated FAK, activated Akt, and active Rac1-GTP. However, TCS possessed no significant effect on activated RhoA level. These results suggested that TCS-induced EMT promoted the motility of anoikis resistant H460 cells through the activation of FAK/Akt signaling pathway as well as Rac1 activation.


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

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

Effects of TCS-mediated EMT on migratory-related proteins.(A) Anoikis resistant H460 cells were treated with TCS at 0–7.5 µM for 24 h in detached condition and then attached on conventional culture dishes for 4 h. The level of pFAK (Tyr 397), FAK, pAkt (Ser 473), Akt, activated Rac1 (Rac1-GTP) and activated RhoA (RhoA-GTP) were determined by western blotting. Blots were reprobed with β-actin to confirm equal loading. (B) The immunoblot signals were quantified by densitometry and mean data from independent experiments were normalized to the results. Values are means of the three triplicate independent samples ± SE. *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-g006: Effects of TCS-mediated EMT on migratory-related proteins.(A) Anoikis resistant H460 cells were treated with TCS at 0–7.5 µM for 24 h in detached condition and then attached on conventional culture dishes for 4 h. The level of pFAK (Tyr 397), FAK, pAkt (Ser 473), Akt, activated Rac1 (Rac1-GTP) and activated RhoA (RhoA-GTP) were determined by western blotting. Blots were reprobed with β-actin to confirm equal loading. (B) The immunoblot signals were quantified by densitometry and mean data from independent experiments were normalized to the results. Values are means of the three triplicate independent samples ± SE. *P<0.05 versus non-treated control.
Mentions: Furthermore, the down-stream effector proteins which are responsible for cell motility were determined using western blotting. The cells were treated with indicated concentrations of TCS for 24 h and subjected to western blot analysis. The expression levels of migratory-related proteins including activated FAK (phosphorylated FAK, Tyr 397), FAK, activated Akt (phosphorylated Akt, Ser 473), Akt, activated Rac1 (Rac1-GTP), and activated RhoA (RhoA-GTP) were investigated. Figures 6A and B show that TCS treatment significantly increased the level of phosphorylated FAK, activated Akt, and active Rac1-GTP. However, TCS possessed no significant effect on activated RhoA level. These results suggested that TCS-induced EMT promoted the motility of anoikis resistant H460 cells through the activation of FAK/Akt signaling pathway as well as Rac1 activation.

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