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5-Lipoxygenase and cysteinyl leukotriene receptor 1 regulate epidermal growth factor-induced cell migration through Tiam1 upregulation and Rac1 activation.

Magi S, Takemoto Y, Kobayashi H, Kasamatsu M, Akita T, Tanaka A, Takano K, Tashiro E, Igarashi Y, Imoto M - Cancer Sci. (2014)

Bottom Line: In this study, we found that 5-lipoxygenase (5-LOX) is activated in the process of EGF-induced cell migration, and that leukotriene C4 (LTC4 ) produced by 5-LOX mediated the second wave of Rac1 activation, as well as cell migration.Furthermore, these effects caused by LTC4 were found to be blocked in the presence of the antagonist of cysteinyl leukotriene receptor 1 (CysLT1).We also found that 5-LOX inhibitors, CysLT1 antagonists and the knockdown of CysLT1 inhibited EGF-induced T cell lymphoma invasion and metastasis-inducing protein 1 (Tiam1) expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Biosciences and Informatics, Faculty of Science and Technology, Keio University, Yokohama, Japan.

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5-lipoxygenase (5-LOX) inhibitors decrease the second epidermal growth factor (EGF)-induced wave of lamellipodia formation and leukotriene synthesis in A431 cells. (a, b) A431 cells were pretreated with the indicated concentrations of compounds for 15 min and stimulated with EGF. After 5 min (first wave; a) or 12 h (second wave; b), the cells were observed by confocal microscopy. The data represent the mean ± SD (n = 3). (c) A431 cells were pretreated with various concentrations of BU-4664L in the presence or absence of LTB4, LTC4 or LTD4 for 15 min, and stimulated with 30 ng/mL of EGF. After 12 h incubation, cells were observed by confocal microscopy. The data represent the mean ± SD (n = 3). (d) Time course experiments of LTC4 production induced by EGF. A431 cells were stimulated with EGF for the indicated periods, and then extracellular levels of LTC4 were measured. The data represent the mean ± SD (n = 3). (e) The effect of 5-LOX inhibitors on the EGF-induced production of LTC4. A431 cells were pretreated with each inhibitor for 15 min and stimulated with EGF for 12 h. Subsequently, the extracellular LTC4 were measured. The data represent the mean ± SD (n = 3).
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fig02: 5-lipoxygenase (5-LOX) inhibitors decrease the second epidermal growth factor (EGF)-induced wave of lamellipodia formation and leukotriene synthesis in A431 cells. (a, b) A431 cells were pretreated with the indicated concentrations of compounds for 15 min and stimulated with EGF. After 5 min (first wave; a) or 12 h (second wave; b), the cells were observed by confocal microscopy. The data represent the mean ± SD (n = 3). (c) A431 cells were pretreated with various concentrations of BU-4664L in the presence or absence of LTB4, LTC4 or LTD4 for 15 min, and stimulated with 30 ng/mL of EGF. After 12 h incubation, cells were observed by confocal microscopy. The data represent the mean ± SD (n = 3). (d) Time course experiments of LTC4 production induced by EGF. A431 cells were stimulated with EGF for the indicated periods, and then extracellular levels of LTC4 were measured. The data represent the mean ± SD (n = 3). (e) The effect of 5-LOX inhibitors on the EGF-induced production of LTC4. A431 cells were pretreated with each inhibitor for 15 min and stimulated with EGF for 12 h. Subsequently, the extracellular LTC4 were measured. The data represent the mean ± SD (n = 3).

Mentions: We first investigated the effect of 5-LOX inhibitors on cell migration following cytoskeletal remodeling. Earlier reports show that EGF-induced actin remodeling is regulated by 5-LOX and its products in epidermoid carcinoma A431cells.21 As shown in Figure1, BU-4664L24,25 and AA-861 inhibited EGF-induced cell migration of A431 cells at IC50 values of 0.66 μg/mL and 9.0 μM, respectively, without affecting cell viability. We previously found that EGF induced two waves of lamellipodia formation, at 5 min and 12 h after stimulation;6 therefore, we evaluated the effect of these 5-LOX inhibitors on each wave of lamellipodia formation. We found that BU-4664L and AA-861 did not inhibit the first wave of lamellipodia formation (Fig.2a), but they inhibited the second wave at IC50 values of 0.69 μg/mL and 11.5 μM, respectively (Fig.2b). These results indicate that 5-LOX played an important role in the second EGF-induced wave of lamellipodia formation in A431 cells.


5-Lipoxygenase and cysteinyl leukotriene receptor 1 regulate epidermal growth factor-induced cell migration through Tiam1 upregulation and Rac1 activation.

Magi S, Takemoto Y, Kobayashi H, Kasamatsu M, Akita T, Tanaka A, Takano K, Tashiro E, Igarashi Y, Imoto M - Cancer Sci. (2014)

5-lipoxygenase (5-LOX) inhibitors decrease the second epidermal growth factor (EGF)-induced wave of lamellipodia formation and leukotriene synthesis in A431 cells. (a, b) A431 cells were pretreated with the indicated concentrations of compounds for 15 min and stimulated with EGF. After 5 min (first wave; a) or 12 h (second wave; b), the cells were observed by confocal microscopy. The data represent the mean ± SD (n = 3). (c) A431 cells were pretreated with various concentrations of BU-4664L in the presence or absence of LTB4, LTC4 or LTD4 for 15 min, and stimulated with 30 ng/mL of EGF. After 12 h incubation, cells were observed by confocal microscopy. The data represent the mean ± SD (n = 3). (d) Time course experiments of LTC4 production induced by EGF. A431 cells were stimulated with EGF for the indicated periods, and then extracellular levels of LTC4 were measured. The data represent the mean ± SD (n = 3). (e) The effect of 5-LOX inhibitors on the EGF-induced production of LTC4. A431 cells were pretreated with each inhibitor for 15 min and stimulated with EGF for 12 h. Subsequently, the extracellular LTC4 were measured. The data represent the mean ± SD (n = 3).
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Related In: Results  -  Collection

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fig02: 5-lipoxygenase (5-LOX) inhibitors decrease the second epidermal growth factor (EGF)-induced wave of lamellipodia formation and leukotriene synthesis in A431 cells. (a, b) A431 cells were pretreated with the indicated concentrations of compounds for 15 min and stimulated with EGF. After 5 min (first wave; a) or 12 h (second wave; b), the cells were observed by confocal microscopy. The data represent the mean ± SD (n = 3). (c) A431 cells were pretreated with various concentrations of BU-4664L in the presence or absence of LTB4, LTC4 or LTD4 for 15 min, and stimulated with 30 ng/mL of EGF. After 12 h incubation, cells were observed by confocal microscopy. The data represent the mean ± SD (n = 3). (d) Time course experiments of LTC4 production induced by EGF. A431 cells were stimulated with EGF for the indicated periods, and then extracellular levels of LTC4 were measured. The data represent the mean ± SD (n = 3). (e) The effect of 5-LOX inhibitors on the EGF-induced production of LTC4. A431 cells were pretreated with each inhibitor for 15 min and stimulated with EGF for 12 h. Subsequently, the extracellular LTC4 were measured. The data represent the mean ± SD (n = 3).
Mentions: We first investigated the effect of 5-LOX inhibitors on cell migration following cytoskeletal remodeling. Earlier reports show that EGF-induced actin remodeling is regulated by 5-LOX and its products in epidermoid carcinoma A431cells.21 As shown in Figure1, BU-4664L24,25 and AA-861 inhibited EGF-induced cell migration of A431 cells at IC50 values of 0.66 μg/mL and 9.0 μM, respectively, without affecting cell viability. We previously found that EGF induced two waves of lamellipodia formation, at 5 min and 12 h after stimulation;6 therefore, we evaluated the effect of these 5-LOX inhibitors on each wave of lamellipodia formation. We found that BU-4664L and AA-861 did not inhibit the first wave of lamellipodia formation (Fig.2a), but they inhibited the second wave at IC50 values of 0.69 μg/mL and 11.5 μM, respectively (Fig.2b). These results indicate that 5-LOX played an important role in the second EGF-induced wave of lamellipodia formation in A431 cells.

Bottom Line: In this study, we found that 5-lipoxygenase (5-LOX) is activated in the process of EGF-induced cell migration, and that leukotriene C4 (LTC4 ) produced by 5-LOX mediated the second wave of Rac1 activation, as well as cell migration.Furthermore, these effects caused by LTC4 were found to be blocked in the presence of the antagonist of cysteinyl leukotriene receptor 1 (CysLT1).We also found that 5-LOX inhibitors, CysLT1 antagonists and the knockdown of CysLT1 inhibited EGF-induced T cell lymphoma invasion and metastasis-inducing protein 1 (Tiam1) expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Biosciences and Informatics, Faculty of Science and Technology, Keio University, Yokohama, Japan.

Show MeSH
Related in: MedlinePlus