阻断NF-κB和ERK通路对黑素瘤细胞增殖及凋亡的影响

发布时间：2018-08-20 13:06

【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(P0.01),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(P0.01)。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。
[Abstract]:Objective to investigate the effects of blocking nuclear factor 魏 B (NF- 魏 B) and extracellular signal-regulated kinase (ERK) on proliferation and apoptosis of melanoma A375 cells. Methods the cultured melanoma cells A375 were randomly divided into two groups: the control group treated with BMS-345541 and the BMS-345541 U0126 treated with BMS-345541 U0126. The proliferation inhibition rate of each group was measured by thiazolyl blue (MTT) assay. Flow cytometry was used to detect the cell cycle and apoptosis rate. Western blot was used to detect the expression of anti-apoptotic protein Bcl-2 in the cells treated with two reagents. Results BMS-345541 and U0126 significantly inhibited the proliferation of A375 cells. The inhibitory rate of proliferation in BMS-345541 U0126 treatment group was (77.27 卤2.70) at 24h, which was significantly higher than that in BMS-345541 group (35.41 卤1.38)% and U0126 group (30.64 卤2.86). The apoptotic rate was increased by 13.96% and 38.91%, respectively. BMS-345541 and U0126 could decrease the expression of Bcl-2 protein in A375 cells. The expression of Bcl-2 protein decreased significantly after combined treatment. All the above differences were statistically significant (P0.01). Conclusion blocking NF- 魏 B and ERK signaling pathways can inhibit the proliferation and induce apoptosis of A375 cells.
【作者单位】：青岛市市立医院皮肤科;青岛大学附属医院皮肤科;
【基金】：山东省自然科学基金(ZR2010HM022) 山东省科技发展计划项目(2011YD18012)
【分类号】：R739.5

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