CAJ | 학술논문

目的:研究表皮生长因子(epidermal growth factor,EGF)诱导小鼠黑素瘤B16细胞移行及其信号通路机制.方法:蛋白质免疫印迹方法分析各蛋白的表达;细胞迁移试验(phagokinetic track motility)测定法观察细胞的迁移.结果:EGF可诱导表皮生长因子受体(EGFR)和细胞外信号调节激酶(ERK)磷酸化,EGF处理后5 min,EGFR和ERK的磷酸化达到峰值,EGFR激酶抑制剂(PD153035)和ERK抑制剂(U0126)可阻断EGF诱导的ERK磷酸化;EGF可显著提高细胞中水通道蛋白-3(aqua-porins-3,AQP3)的活性,并随时间延长活性增强,24 h达到较高水平,AQP3活性亦随浓度升高而增强,当EGF浓度为100μg/L时达到较高水平:PD153035、U0126和CuSO_4可抑制AQP3的活性以及细胞的移行.结论:在小鼠黑素瘤B16细胞中,EGF通过磷酸化EGFR,激活ERK,进而使AQP3表达上调,促进B16细胞迁移.这一通路可被EGFR激酶抑制剂、ERK和AQP3抑制剂阻断,这些涉及的信号通路可能形成潜在的治疗黑素瘤的靶点.
목적:연구표피생장인자(epidermal growth factor,EGF)유도소서흑소류B16세포이행급기신호통로궤제.방법:단백질면역인적방법분석각단백적표체;세포천이시험(phagokinetic track motility)측정법관찰세포적천이.결과:EGF가유도표피생장인자수체(EGFR)화세포외신호조절격매(ERK)린산화,EGF처리후5 min,EGFR화ERK적린산화체도봉치,EGFR격매억제제(PD153035)화ERK억제제(U0126)가조단EGF유도적ERK린산화;EGF가현저제고세포중수통도단백-3(aqua-porins-3,AQP3)적활성,병수시간연장활성증강,24 h체도교고수평,AQP3활성역수농도승고이증강,당EGF농도위100μg/L시체도교고수평:PD153035、U0126화CuSO_4가억제AQP3적활성이급세포적이행.결론:재소서흑소류B16세포중,EGF통과린산화EGFR,격활ERK,진이사AQP3표체상조,촉진B16세포천이.저일통로가피EGFR격매억제제、ERK화AQP3억제제조단,저사섭급적신호통로가능형성잠재적치료흑소류적파점.
Objective: To investigate the migration induced by epidermal growth factor (EGF) and its signal pathway in cultured mouse melanoma B16 cells. Methods:Cultured B16 cells were treated with EGF and/or various reagents and subjected to cell migration assay by phagokinetic track motility or biochemical analysis for expression or activation of proteins by SDS-PAGE/Westem blot analysis. Results:Aquaporins-3 (AQP3) was expressed in cultured mouse melanoma B16 cells. AQP3 expression and migration were up-regulated by EGF. EGF-induced cell migration was inhibited by AQP3 water transport inhibitor CuSO_4, and the effects of EGF mediated, at least in part, by its inhibitory effects on EGFR and downstream ERK activation. Conclusions:EGF enhances AQP3 expression and cell migration in cultured mouse melanoma B16 cells. EGFR/ERK activation is involved in this process. These results may provide a new potential therapeutic target in melanoma.