外源性S100A6对黑色素瘤B16细胞增殖、凋亡的影响及其机制  

Effect of exogenous S100A6 on proliferation and apoptosis of melanoma B16 cells and relevant mechanism

作　　者：陈英华[1] 孙双双[1] 卫佳[1] 李星星[1] 游莉[1] 邹正渝[1] 黎玉叶[1] 何通川[1] 周兰[1] 

机构地区：[1]重庆医科大学检验医学院临床检验诊断学教育部重点实验室,重庆400016

出　　处：《中国生物制品学杂志》2012年第3期304-308,共5页Chinese Journal of Biologicals

基　　金：国家自然科学基金资助(30772548)

摘　　要：目的探讨外源性S100A6对黑色素瘤B16细胞增殖、凋亡的影响及其机制。方法采用MTT、台盼蓝拒染、Hoechst、Western blot、免疫细胞化学、免疫荧光和荧光素酶活性分析法检测S100A6对B16细胞增殖、凋亡、β-catenin的水平和分布、β-catenin/TCF4转录活性及经典Wnt信号途径(即Wnt/β-catenin信号)的下游靶基因c-myc表达的影响。结果 GST-hS100A6可抑制B16细胞的增殖,且呈时间-剂量依赖性;GST-hS100A6作用72 h后,B16细胞的凋亡率较GST对照组增加1.43倍;携带S100A6基因的重组腺病毒AdS100A6作用72 h后,B16细胞中β-catenin的表达量较对照腺病毒AdGFP组增加43.9%;GST-hS100A6可使B16细胞中β-catenin的表达增加,且以胞核增加为主;GST-hS100A6作用后,B16细胞的β-catenin/TCF4转录活性增至GST对照组的7.4倍,且该信号途径的靶基因之一c-myc的表达也增加70.4%。上述结果与GST对照组之间的差异均有统计学意义(P均<0.05或0.01)。结论 S100A6具有抑制黑色素瘤增殖、促进凋亡和上调其Wnt/β-catenin信号途径活性的作用,且上调Wnt/β-catenin信号途径活性可能是S100A6对黑色素瘤抑制性作用的机制之一。Objective To investigate the effect of exogenous S100A6 on proliferation and apoptosis of melanoma B16 cells as well as the relevant mechanism.Methods The effects of S100A6 on the proliferation and apoptosis of B16 cells,the level and distribution of β-catenin,the transcription activity of β-catenin / TCF4 as well as expression of downstream target gene c-myc of typical Wnt（Wnt / β-catenin） signaling pathway were investigated by MTT method,trypan blue dye exclusion assay,Hoechst staining,Western blot,immunocytochemical assay,IFA and luciferase activity assay.Results GST-hS100A6 showed time-and dose-dependent inhibitory effect on proliferation of B16 cells.The apoptosis rate of B16 cells 72 h after treatment with GST-hS100A6 increased by 1.43 folds as compared with those after treatment with GST.The expression level of β-catenin in B16 cells 72 h after treatment with recombinant adenovirus AdS100A6 carrying S100A6 gene increased by 43.9% as compared with those after treatment with control adenovirus AdGFP.GST-hS100A6 increased the expression of β-catenin in B16 cells especially in nucleus.The transcription activity of β-catenin / TCF4 in B16 cells after treatment with GST-hS100A6 was 7.4 times of that in control cells,while the expression level of c-myc gene increased by 70.4%.All the results showed significant difference with those in GST control group（P 0.05 or P 0.01）.Conclusion S100A6 inhibited the proliferation and promoted the apoptosis of melanoma cells by possible mechanism of up-regulating the activity of Wnt / β-catenin signaling pathway.

关 键 词：S100蛋白质类 黑色素瘤 细胞增殖 细胞凋亡 Wnt/β-catenin信号途径 

分 类 号：R739.5[医药卫生—肿瘤]