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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 on anchorage-independent growth of anoikis resistant H460 cells.(A) Cells were pretreated with TCS (0–7.5 µM) for 24 h and subjected to soft agar colony formation assay, as described in “Materials and Methods”. Representative fields from three independent experiments were photographed after the cells were cultured for 7 and 10 days. Scale bar is 1,000 µm. (B–C) Colony number and colony size were determined by image analyzer on the 10th day of culture. Values are means of the three independent triplicate samples ± SE. *P<0.05 versus non-treated control.
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pone-0110851-g002: Effects of TCS on anchorage-independent growth of anoikis resistant H460 cells.(A) Cells were pretreated with TCS (0–7.5 µM) for 24 h and subjected to soft agar colony formation assay, as described in “Materials and Methods”. Representative fields from three independent experiments were photographed after the cells were cultured for 7 and 10 days. Scale bar is 1,000 µm. (B–C) Colony number and colony size were determined by image analyzer on the 10th day of culture. Values are means of the three independent triplicate samples ± SE. *P<0.05 versus non-treated control.

Mentions: Because anchorage-independent growth of the cancer cells has been shown to augment metastasis, we next investigated the effect of TCS on cancer cell growth in such condition. In doing this, anoikis resistant cells H460 were treated with TCS for 24 h before they were subjected to colony formation assay. The cells were then seeded in agarose layer to prevent cell-cell interaction and attachment. Colony number and colony size were obtained by photographing and counting after the cells were cultured for 7 and 10 days. The colony formation was shown in Figure 2A. Colony number and colony size of each treatment were calculated as a percentage of the control group and shown in Figure 2B and C, respectively. The results indicated that TCS at the concentrations of 5 and 7.5 µM significantly increased colony formation of anoikis resistant H460 cells. However, TCS at both concentrations significantly reduced colony size in comparison to that of non-treated control group. Such observations indicated that TCS promoted anchorage-independent survival of the cells, but decreased the growth rate of the cells in detached condition. Our results consists with the previous findings that the increase in anchorage-independent survival with low proliferative ability has been observed in the cancer cells undergoing EMT [11], [12], [41], [42].


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 on anchorage-independent growth of anoikis resistant H460 cells.(A) Cells were pretreated with TCS (0–7.5 µM) for 24 h and subjected to soft agar colony formation assay, as described in “Materials and Methods”. Representative fields from three independent experiments were photographed after the cells were cultured for 7 and 10 days. Scale bar is 1,000 µm. (B–C) Colony number and colony size were determined by image analyzer on the 10th day of culture. Values are means of the three independent triplicate 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-g002: Effects of TCS on anchorage-independent growth of anoikis resistant H460 cells.(A) Cells were pretreated with TCS (0–7.5 µM) for 24 h and subjected to soft agar colony formation assay, as described in “Materials and Methods”. Representative fields from three independent experiments were photographed after the cells were cultured for 7 and 10 days. Scale bar is 1,000 µm. (B–C) Colony number and colony size were determined by image analyzer on the 10th day of culture. Values are means of the three independent triplicate samples ± SE. *P<0.05 versus non-treated control.
Mentions: Because anchorage-independent growth of the cancer cells has been shown to augment metastasis, we next investigated the effect of TCS on cancer cell growth in such condition. In doing this, anoikis resistant cells H460 were treated with TCS for 24 h before they were subjected to colony formation assay. The cells were then seeded in agarose layer to prevent cell-cell interaction and attachment. Colony number and colony size were obtained by photographing and counting after the cells were cultured for 7 and 10 days. The colony formation was shown in Figure 2A. Colony number and colony size of each treatment were calculated as a percentage of the control group and shown in Figure 2B and C, respectively. The results indicated that TCS at the concentrations of 5 and 7.5 µM significantly increased colony formation of anoikis resistant H460 cells. However, TCS at both concentrations significantly reduced colony size in comparison to that of non-treated control group. Such observations indicated that TCS promoted anchorage-independent survival of the cells, but decreased the growth rate of the cells in detached condition. Our results consists with the previous findings that the increase in anchorage-independent survival with low proliferative ability has been observed in the cancer cells undergoing EMT [11], [12], [41], [42].

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