阻断NF-κB和ERK通路对黑素瘤细胞增殖及凋亡的影响  

The Effects of Blocking NF-κB and ERK Signaling Transduction Pathway on the Proliferation and Apopatosis in Human Melanoma Cell Line A375

作　　者：刘宁[1] 肖君刚[2] 潘敏[2] 郁博[2] 

机构地区：[1]青岛市市立医院皮肤科,山东青岛266000 [2]青岛大学附属医院皮肤科,山东青岛266003

出　　处：《中国皮肤性病学杂志》2014年第7期667-671,共5页The Chinese Journal of Dermatovenereology

基　　金：山东省自然科学基金(ZR2010HM022);山东省科技发展计划项目(2011YD18012)

摘　　要：目的研究分别或联合阻断核因子κB(NF-κB)和细胞外信号调节激酶ERK两条信号通路对黑素瘤A375细胞增殖及凋亡的影响。方法将培养的黑素瘤细胞A375随机分为对照组、NF-κB通路抑制剂BMS-345541组、ERK通路抑制剂U0126处理组及BMS-345541+U0126处理组,采用噻唑蓝法(MTT)测定各组细胞的增殖抑制率;流式细胞术检测各组细胞周期及凋亡率;Western blot检测两试剂处理后细胞中抗凋亡蛋白Bcl-2的表达。结果 BMS-345541和U0126作用后对A375细胞增殖有明显抑制作用,且呈浓度和时间依赖性(P<0.01),作用时间24h时,BMS-345541+U0126处理组的增殖抑制率为(77.27±2.70)%,明显高于BMS-345541组(35.41±1.38)%及U0126组(30.64±2.86)%,且联合作用具有协同性;BMS-345541组、U0126组和BMS-345541+U0126处理组与对照组相比S期分别降低14.20%,18.40%及22.64%,而凋亡率分别提高24.98%,13.96%及38.91%;BMS-345541和U0126均可降低A375细胞中Bcl-2蛋白的表达,二者联合作用后Bcl-2蛋白的表达显著降低。以上差异均有统计学意义(P<0.01)。结论阻断NF-κB和ERK两条信号通路对A375细胞具有增殖抑制和诱导凋亡作用,两者联合具有协同效应。Objective To investigate the effects of blocking the NF-KB and ERK signaling pathway on proliferation and apoptosis in human melanoma cell line A375 . Methods Cultured A375 cells were randomly classified into 4 groups ： control group, BMS-345541 （ NF-KB inhibitor） group, U0126 （ ERK inhibitor） group and BMS-345541 ＋ U0126 group,respectively. The proliferation inhibition rate of cells was measured by MTT method. Flow cytometry was performed to detect the cell cycle and apoptosis rate of A375 cells. The protein expression of Bcl-2 was detected by Western blot. Results The proliferation inhibition rate on A375 cells treated by BMS-345541, U0126 and combination group after 24h was （35.41 ± 1.38 ）%, （30.64± 2.86） % and （ 77.27 ± 2.70 ） % （ P 〈 0.01 ）, respectively. The inhibition of proliferation was in a dose and time dependent fashion. Comparing to the control group, phase S cells number decreased 14.20%, 18.40% and 22.64% in BMS-345541 group, U0126 group and combination group respectively （P 〈 0. 01 ）. The apoptosis rate of BMS-345541 group , U0126 group and combination group was increased （24.98±1.03）% ,（13.96 ±0.96）% and （38.91 ±1.46）% ,in contrast with the control group （P〈 0. 01 ）. Both BMS-345541 and U0126 can remarkably induce the expression of Bcl-2 in cell line A375. BMS-345541 combined with U0126 is more effective （ P 〈 0.01 ）. Conclusion Blocking NF-kB and ERK signaling pathway can inhibit the cell proliferation and induce apoptosis, and the combination of them exerts a synergistic effect in the cell line A375.

关 键 词：NF-KB ERK 黑素瘤细胞 

分 类 号：R739.5[医药卫生—肿瘤]