Isogarcinol对人红白血病细胞HEL的抗肿瘤作用及机制  

The Effect and Mechanism of Isogarcinol on Apoptosis and Proliferation of Human Erythroleukemia Cell Line HEL

作　　者：陈丽 杨珏 邱剑飞 苑春茂 吴昌学[3] 李艳梅 郝小江 CHEN Li;YANG Jue;QIU Jianfei;YUAN Chunmao;WU Changxue;LI Yanmei;HAO Xiaojiang(Guizhou University of Chinese Medicine,Guiyang 550002,Guizhou,China;The Key Laboratory of Chemistry for Natural Products of Guizhou Province and Chinese Academic of Sciences,Guiyang 550014,Guizhou,China;Guizhou Medical University,Guiyang 550025,Guizhou,China)

机构地区：[1]贵州中医药大学,贵州贵阳550002 [2]贵州省中科院天然产物化学重点实验室,贵州贵阳550014 [3]贵州医科大学,贵州贵阳550025

出　　处：《贵州医科大学学报》2019年第1期35-41,47,共8页Journal of Guizhou Medical University

基　　金：国家自然科学基金(81700169);贵州省科技合作计划项目[黔科合LG字(2012)009]

摘　　要：目的:研究Isogarcinol对人红白血病细胞HEL的凋亡、增殖的影响及其作用机制。方法:采用不同浓度的Isogarcinol化合物处理HEL细胞,分别采用MTT测定细胞活力,Annexin V/PI双染流式细胞术检测细胞凋亡,细胞计数绘制细胞生长曲,Western blot和qRT-PCR法检测胞内信号通路相关基因在蛋白和基因水平的表达,利用SCID白血病小鼠模型体内评价该化合物的作用。结果:Isogarcinol抑制人红白血病细胞HEL的活性,IC505.4μmol/L可显著抑制人红白血病细胞HEL的增殖及诱导细胞凋亡; qRT-PCR结果显示促凋亡基因Bax的表达增加,而Caspase-3、Caspase-8的表达发生下调,抗凋亡基因Bcl-2、c-Myc的表达下调; Western blot结果显示Bad,Bax,Cleaved caspase-3,Cleaved caspase-8和Cleaved PARP的相对表达量呈浓度依赖性上调,而JAK2,PStat3、Bcl-2、Caspase3、Caspase-8、PARP、P-ERK 1/2和c-Myc的相对表达量呈浓度依赖性下调;小鼠体内实验表明Isogarcinol显著延长了小鼠的存活时间,并且增加了血红细胞比。结论:Isogarcinol诱导人红白血病细胞HEL细胞凋亡和抑制HEL细胞增殖,可能通过抑制JAK2/STAT3信号通路而实现的。Objective:To study the effect and mechanism of Isogarcinol on apoptosis and proliferation of human erythroleukemia cell line HEL.Methods:MTT assay was used to measure the inhibition rate of different concentration of Isogarcinol on HEL cells.Apoptosis was detected by Annexin V/PI staining using flow cytometry.Cell growth curve was drawn based on cell counting data.Expressions of intracellular signal pathway-related proteins and genes were detected by Western blot and qRT-PCR.HEL tumor-bearing SCID mice were used to eval uate antitumor effect of this compound.Results:Isogarcinol significantly inhibited HEL cell viability and induced HEL cellular apoptosis,and its IC50 was 5.4μmol/L.qRT-PCR results showed that the expression of proapoptotic gene Bax was increased,while the expression levels of Caspase-3 and Caspase-8 were downregulated.In addition,the expression levels of anti-apoptotic genes Bcl-2 and c-Myc were downregulated.Western blot analysis showed that the expression levels of Bad,Bax,Cleaved caspase-3,Cleaved caspase-8 and Cleaved PARP were upregulated in dose-dependent manner;while the expression levels of JAK2,P-Stat3,Bcl-2,Caspase3,Caspase-8,PARP,P-ERK 1/2 and c-Myc were down-regulated in dose-dependent manner.Moreover,Isogarcinol significantly prolonged the survival of HEL tumor-bearing mice and increased the hematocrit ratio.Conclusion:Isogarcinol induces apoptosis of HEL cells and inhibits HEL cell proliferation,which may be achieved by inhibiting JAK2/STAT3 signaling pathway.

关 键 词：Isogarcinol 人红白血病细胞 细胞增殖 细胞凋亡 JAK2/STAT3信号通路 

分 类 号：R966[医药卫生—药理学]