中华皮肤科杂志
中華皮膚科雜誌
중화피부과잡지
Chinese Journal of Dermatology
2014年
8期
570-573
,共4页
秦绪艳%臧运书%潘敏%袁梦姝%郁博%吴惠惠
秦緒豔%臧運書%潘敏%袁夢姝%鬱博%吳惠惠
진서염%장운서%반민%원몽주%욱박%오혜혜
黑色素瘤%NF-κB%细胞外信号调节MAP激酶类%信号转导
黑色素瘤%NF-κB%細胞外信號調節MAP激酶類%信號轉導
흑색소류%NF-κB%세포외신호조절MAP격매류%신호전도
Melanoma%NF-kappa B%Extracellular signal-regulated MAP kinase%Signal transducting
目的 探讨黑素瘤细胞A375细胞外信号调节激酶(ERK)通路与核因子κB(NF-κB)信号转导通路的交互作用.方法 将培养的A375细胞分为对照组、选择性ERK信号通路抑制剂U0126(10 μmol/L、5 μmol/L)处理组和选择性NF-κB通路抑制剂BMS-345541(10 μmol/L、5μmol/L)处理组,分别提取蛋白质和mRNA,用Western印迹、RT-PCR观察U0126抑制ERK通路后,NF-κB P65、p-IκBα蛋白及NF-κB P65mRNA的表达,观察BMS-345541抑制NF-κB通路后ERK1/2、p-ERK1/2蛋白及ERK1 mRNA的表达.结果 应用10 μmol/L、5μmol/L U0126抑制ERK通路后,胞核NF-κB P65蛋白表达(分别为0.60±0.04、0.56±0.06)均较对照组(1.54±0.15)减少(P＜0.01),其表达量与药物浓度无显著相关性(P＞ 0.01);p-IκBα蛋白的表达(0.90±0.05、0.70±0.02)均较对照组(0.61±0.03)增加(P＜0.01),两药物浓度组之间的表达量差异有统计学意义(P＜ 0.01);NF-κB P65 mRNA的表达(0.79±0.05,0.75±0.04)均较对照组(0.86±0.05)减少(P＜0.01).应用10 μmol/L BMS-345541抑制NF-κB通路后,ERK1/2、p-ERK1/2蛋白及ERK1 mRNA的表达(0.73±0.07、0.75±0.09、1.51±0.02)较对照组减少(P＜0.01),BMS-345541浓度为5μmol/L时,ERK1/2 、p-ERK 1/2蛋白和ERK1 mRNA的表达(0.94±0.11、0.99±0.04、1.62±0.03)较对照组的差异无统计学意义(均P＞0.05).结论 NF-κB通路和ERK通路在黑素瘤A375细胞中存在交互作用.
目的 探討黑素瘤細胞A375細胞外信號調節激酶(ERK)通路與覈因子κB(NF-κB)信號轉導通路的交互作用.方法 將培養的A375細胞分為對照組、選擇性ERK信號通路抑製劑U0126(10 μmol/L、5 μmol/L)處理組和選擇性NF-κB通路抑製劑BMS-345541(10 μmol/L、5μmol/L)處理組,分彆提取蛋白質和mRNA,用Western印跡、RT-PCR觀察U0126抑製ERK通路後,NF-κB P65、p-IκBα蛋白及NF-κB P65mRNA的錶達,觀察BMS-345541抑製NF-κB通路後ERK1/2、p-ERK1/2蛋白及ERK1 mRNA的錶達.結果 應用10 μmol/L、5μmol/L U0126抑製ERK通路後,胞覈NF-κB P65蛋白錶達(分彆為0.60±0.04、0.56±0.06)均較對照組(1.54±0.15)減少(P＜0.01),其錶達量與藥物濃度無顯著相關性(P＞ 0.01);p-IκBα蛋白的錶達(0.90±0.05、0.70±0.02)均較對照組(0.61±0.03)增加(P＜0.01),兩藥物濃度組之間的錶達量差異有統計學意義(P＜ 0.01);NF-κB P65 mRNA的錶達(0.79±0.05,0.75±0.04)均較對照組(0.86±0.05)減少(P＜0.01).應用10 μmol/L BMS-345541抑製NF-κB通路後,ERK1/2、p-ERK1/2蛋白及ERK1 mRNA的錶達(0.73±0.07、0.75±0.09、1.51±0.02)較對照組減少(P＜0.01),BMS-345541濃度為5μmol/L時,ERK1/2 、p-ERK 1/2蛋白和ERK1 mRNA的錶達(0.94±0.11、0.99±0.04、1.62±0.03)較對照組的差異無統計學意義(均P＞0.05).結論 NF-κB通路和ERK通路在黑素瘤A375細胞中存在交互作用.
목적 탐토흑소류세포A375세포외신호조절격매(ERK)통로여핵인자κB(NF-κB)신호전도통로적교호작용.방법 장배양적A375세포분위대조조、선택성ERK신호통로억제제U0126(10 μmol/L、5 μmol/L)처리조화선택성NF-κB통로억제제BMS-345541(10 μmol/L、5μmol/L)처리조,분별제취단백질화mRNA,용Western인적、RT-PCR관찰U0126억제ERK통로후,NF-κB P65、p-IκBα단백급NF-κB P65mRNA적표체,관찰BMS-345541억제NF-κB통로후ERK1/2、p-ERK1/2단백급ERK1 mRNA적표체.결과 응용10 μmol/L、5μmol/L U0126억제ERK통로후,포핵NF-κB P65단백표체(분별위0.60±0.04、0.56±0.06)균교대조조(1.54±0.15)감소(P＜0.01),기표체량여약물농도무현저상관성(P＞ 0.01);p-IκBα단백적표체(0.90±0.05、0.70±0.02)균교대조조(0.61±0.03)증가(P＜0.01),량약물농도조지간적표체량차이유통계학의의(P＜ 0.01);NF-κB P65 mRNA적표체(0.79±0.05,0.75±0.04)균교대조조(0.86±0.05)감소(P＜0.01).응용10 μmol/L BMS-345541억제NF-κB통로후,ERK1/2、p-ERK1/2단백급ERK1 mRNA적표체(0.73±0.07、0.75±0.09、1.51±0.02)교대조조감소(P＜0.01),BMS-345541농도위5μmol/L시,ERK1/2 、p-ERK 1/2단백화ERK1 mRNA적표체(0.94±0.11、0.99±0.04、1.62±0.03)교대조조적차이무통계학의의(균P＞0.05).결론 NF-κB통로화ERK통로재흑소류A375세포중존재교호작용.
Objective To investigate the cross-talk between extracellular signal-regulated kinase (ERK) and nuclear factor κB (NF-κB) signal transduction pathways in A375 human melanoma cells.Methods Cultured A375 cells were randomly divided into 5 groups:control group receiving no treatment,two U0126 (a selective inhibitor of the ERK signaling pathway) groups treated with U0126 of 10 and 5 μmol/L,and two BMS-345541 groups treated with BMS-345541 of 10 and 5 μmol/L.After 24-hour treatment,Western blot and reverse transcription PCR were performed to measure the protein expressions of NF-κB P65,phosphorylated IκBα (p-IκBα),ERK1/2,as well as p-ERK1/2,and the mRNA expressions of NF-κB P65 and ERK1,respectively.One-way analysis of variance and least significant difference (LSD)-t test were carried out for statistical analysis.Results After 24 hours of treatment with U0126 of 10 and 5 μmol/L,a significant decrease was noted in the relative expression level of NF-κB p65 protein (0.60 ± 0.04 and 0.56 ± 0.06 vs.1.54 ± 0.15,both P＜ 0.01) and mRNA (0.79 ± 0.05 and 0.75 ± 0.04 vs.0.86 ± 0.05,both P ＜ 0.01),but a statistical increase in that of p-IκBα protein (0.90 ± 0.05 and 0.70 ± 0.02 vs.0.61 ± 0.03,both P ＜ 0.01) in the two U0126 groups compared with the control group; significant differences were observed in the expression level of p-IκBo protein (P ＜ 0.01) but not in that of NF-κB p65 protein (P ＞ 0.01) between the two U0126 groups.The relative expression levels of ERK1/2 and p-ERK1/2 proteins as well as ERK1 mRNA were significantly higher in the control A375 cells than those in the cells treated with BMS-345541 of 10 μmol/L (0.73 ± 0.07,0.75 ± 0.09,1.51 ± 0.02,all P ＜ 0.01),but similar to those treated with BMS-345541 of 5 μmol/L (0.94 ± 0.11,0.99 ± 0.04,1.62 ± 0.03,all P ＞ 0.05).Conclusion There is a cross-talk between ERK and NF-κB signal transduction pathways in A375 melanoma cells.
