IL-6/JAK/STAT3介导苦参碱对K562细胞的增殖抑制作用  

Growth inhibition effect of matrine on K562 cells mediated by IL-6/JAK/STAT3 signaling pathway

作　　者：马玲娣[1] 朱志超[1] 孙晓[1] 蒋丽佳[1] 白煜[1] 卢绪章[2] 周民[2] 钱思轩[3] 李建勇[3] 

机构地区：[1]南京医科大学附属常州市第二人民医院中心实验室,常州213000 [2]南京医科大学附属常州市第二人民医院血液科,常州213000 [3]南京医科大学附属第一医院、江苏省人民医院血液科

出　　处：《中华血液学杂志》2015年第5期422-426,共5页Chinese Journal of Hematology

基　　金：国家自然科学基金（81101647）;常州市医学卫生创新人才项目;南京医科大学博士后课题

摘　　要：目的 探讨JAK/STAT3介导的信号途径在苦参碱对K562细胞增殖抑制作用中的分子机制.方法 以不同浓度苦参碱处理K562细胞,Western blot法分析JAK/STAT3通路分子JAK2、STAT3及其磷酸化蛋白表达的改变;荧光实时定量RT-PCR及Western blot法分析STAT3下游调控基因Bcl-xL、Cyclin D1、c-Myc的表达;荧光实时定量RT-PCR及ELISA法检测K562细胞IL-6表达的改变.以IL-6预处理K562细胞,Western blot法分析苦参碱对细胞内JAK2、STAT3及其磷酸化蛋白表达的作用.结果 0.5 mg/ml苦参碱处理48 h,K562细胞内STAT3及JAK2总蛋白表达无明显变化,磷酸化STAT3（p-Tyr705 STAT3、p-Ser727 STAT3）和磷酸化JAK2 （p-JAK2）表达水平分别由处理前的0.690±0.119、1.150±0.263和0.670±0.137下降至0.370±0.024、0.700±0.172和0.049±0.057.苦参碱对STAT3下游调控基因Bcl-xL、Cyclin D1及c-Myc的转录及蛋白表达均有抑制作用,并可显著抑制IL-6的表达和分泌,0.5和0.8 mg/ml苦参碱处理后细胞培养上清中IL-6浓度分别为（10.74±1.83）pg/ml和（8.66±1.24） pg/ml,较处理前[（35.1±1.93） pg/ml]显著降低（P＜0.05）.IL-6预处理K562细胞后,STAT3、JAK2、p-STAT3及p-JAK2表达均有增加,苦参碱可抑制IL-6对上述分子表达的上调.结论 苦参碱抑制K562细胞增殖与JAK/STAT3介导的信号通路抑制有关,IL-6表达抑制可能是苦参碱调控JAK/STAT3通路活性的始动机制.Objective To investigate the molecular mechanism of the growth inhibitory effect of matrine on K562 cells in JAK/STAT3 mediated signal pathway.Methods Western blot analyses were performed to investigate the differential expression of JAK2,STAT3,phospho-STAT3 （Tyr705 ＆amp; Ser727） and phospho-JAK2 proteins after matrine treatment in K562 cells with or without human recombinant interleukin 6 （IL-6） pretreatment.The expressions of STAT3 response gene products such as Bcl-xL,Cyclin D1 and c-Myc,were investigated by Western blot and quantitative real time RT-PCR （qRT-PCR）.Expression of IL-6,a potent upstream activating factor of JAK/STAT3 pathway,was analyzed by both real time qRT-PCR and ELISA.Results Western blot revealed that matrine treatment resulted in a strong down-regulation of phospho-STAT3 both in Tyr705 and Ser727 sites or phospho-JAK2 proteins expression without significant effects on the total STAT3 and JAK2 proteins.The expression of phospho-Tyr705 STAT3 and phospho-Ser727 STAT3 was decreased to 0.370±0.024 and 0.700±0.172 in K562 cells treated with 0.5 mg/ml matrine for 48 h,respectively,from 0.690±0.119 and 1.150±0.263 in control cells,accompanied with a dramatical down-regulation of phospho-JAK2 from 0.670±0.137 to 0.049±0.057 （P〈 0.05）.In addition,it was found that the expression of Bcl-xL,Cyclin D1,c-Myc was decreased both at the transcriptional and protein level in K562 cells after matrine treatment.Matrine treatment resulted in a significant decrease in the expression level of IL-6 in K562 cells from （35.1± 1.93） to （10.74±1.83） and （8.66± 1.24） pg/ml at the dose of 0.5 and 0.8 mg/ml,respectively （P〈0.05）.Matrine treatment could diminish the up-regulation of STAT3,JAK2,phospho-STAT3 and phospho-JAK2 protein following pretreatment with IL-6 in K562 cells.Conclusion Matrine exerts its anti-leukemia effect by interfering with the JAK2/STAT3 signaling pathway.The inhibition of IL-6 expression may play a pivotal role in the disruption of JAK/STAT pathway b

关 键 词：苦参碱 K562细胞 白细胞介素6 JAK/STAT3信号通路 

分 类 号：R285[医药卫生—中药学]