贵州医科大学学报

2019, v.44;No.220(01) 35-41+47

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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

陈丽;杨珏;邱剑飞;苑春茂;吴昌学;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;The Key Laboratory of Chemistry for Natural Products of Guizhou Province and Chinese Academic of Sciences;Guizhou Medical University;

摘要(Abstract):

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 evaluate antitumor effect of this compound.Results:Isogarcinol significantly inhibited HEL cell viability and induced HEL cellular apoptosis,and its IC50 was 5.4 μmol/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(KeyWords): Isogarcinol;人红白血病细胞;细胞增殖;细胞凋亡;JAK2/STAT3信号通路
isogarcinol;human erythroleukemia cell line;cell proliferation;apoptosis;JAK2/STAT3 signaling pathway

Abstract:

Keywords:

基金项目(Foundation): 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]

作者(Author): 陈丽;杨珏;邱剑飞;苑春茂;吴昌学;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;The Key Laboratory of Chemistry for Natural Products of Guizhou Province and Chinese Academic of Sciences;Guizhou Medical University;

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DOI: 10.19367/j.cnki.1000-2707.2019.01.007

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