CAJ | 학술논문

目的:研究XIAP抑制剂Embelin体外对人T淋巴瘤细胞Jurkat增殖的影响,并探讨其作用机制.方法:应用MTS法分析不同浓度Embelin对人T淋巴瘤细胞增殖的影响;光学显微镜下观察经Embelin处理后细胞形态学的改变;经Annexin V/PI双染后用流式细胞术检测 Embelin对Jurkat细胞凋亡的影响;Western blot方法检测Embelin 作用后细胞XIAP、PARP、Caspase-3、Cas?pase-8、Caspase-9、促凋亡蛋白Bax和抗凋亡蛋白Bcl-xl、Bcl-2表达的变化.结果:Emebeli对人白血病细胞具有显著增殖抑制作用(P<0.05);Caspase-3抑制剂z-DEVD-fmk,Caspase-9抑制剂Ac-LEHD-CHO能显著下调这种增殖抑制作用.经不同浓度Eme?belin处理24 h后,Jurkat细胞凋亡率明显增高,与未处理组比较有显著性差异(P<0.01);经Emebelin处理24 h后,Jurkat 细胞出现PARP、Caspase3、9裂解片段,且Bax 蛋白表达上调,Bcl-2和Bcl-xL蛋白表达水平下调.结论:Embelin在体外可明显抑制人T淋巴瘤细胞 Jurkat 的增殖,诱导细胞凋亡,其机制可能是通过拮抗 XIAP 的作用,激活 Caspase 依赖性的细胞凋亡内源途径而诱导Jurkat细胞发生凋亡.
목적:연구XIAP억제제Embelin체외대인T림파류세포Jurkat증식적영향,병탐토기작용궤제.방법:응용MTS법분석불동농도Embelin대인T림파류세포증식적영향;광학현미경하관찰경Embelin처리후세포형태학적개변;경Annexin V/PI쌍염후용류식세포술검측 Embelin대Jurkat세포조망적영향;Western blot방법검측Embelin 작용후세포XIAP、PARP、Caspase-3、Cas?pase-8、Caspase-9、촉조망단백Bax화항조망단백Bcl-xl、Bcl-2표체적변화.결과:Emebeli대인백혈병세포구유현저증식억제작용(P<0.05);Caspase-3억제제z-DEVD-fmk,Caspase-9억제제Ac-LEHD-CHO능현저하조저충증식억제작용.경불동농도Eme?belin처리24 h후,Jurkat세포조망솔명현증고,여미처리조비교유현저성차이(P<0.01);경Emebelin처리24 h후,Jurkat 세포출현PARP、Caspase3、9렬해편단,차Bax 단백표체상조,Bcl-2화Bcl-xL단백표체수평하조.결론:Embelin재체외가명현억제인T림파류세포 Jurkat 적증식,유도세포조망,기궤제가능시통과길항 XIAP 적작용,격활 Caspase 의뢰성적세포조망내원도경이유도Jurkat세포발생조망.
Objective: This work aimed to determine the effect of the XIAP inhibitor embelin on the proliferation of human acute T-cell lymphoblastic leukemia in Jurkat cells and to elucidate the mechanisms involved. Methods: The growth suppression of Jurkat cells by embelin was analyzed using the MTS [3(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt] assay. The morphological changes of the cancer cells were observed by light microscopy. The apoptotic rate was evaluated by flow cytometry after annexin/propidium iodide double staining. Western blot analysis was performed to assess the expression of the X-linked inhibitor of apoptosis (XIAP), poly adenosine diphosphate diphosphate phosphoribosyl transferase (PARP), caspase-3, caspase-8, caspase-9, and the pro-apoptotic factor Bax as well as the pro-survival factors Bcl-xL and Bcl-2. Results: Embelin significantly inhibited the proliferation of the human leukemia cells (P<0.05). However, the caspase-3 and caspase-9 inhibitors, z-DEVD-fmk and Ac-LEHD-CHO, respectively, could reverse the action of embelin. The apoptosis of Jurkat cells was significantly increased after 24 h of embelin treatment as compared with that of the control group (P<0.05). Furthermore, the cleavage of PARP, caspase-3, and caspase-9 was observed (P<0.05). Moreover, Bax expression was upregulated while Bcl-2 and Bcl-xL levels were decreased. Conclusion: Embelin can significantly inhibit the proliferation of human T-cell lymphoma in the Jurkat cell line by antagonizing the endogenous XIAP pathway that activates caspase-dependent apoptosis in Jurkat cells.