Isogarcinol对人红白血病细胞HEL的抗肿瘤作用及机制

摘要

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.%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.