摘要
目的:研究辣椒素(capsaicin)是否经内质网途径诱导耐药性白血病K562/ADM细胞凋亡。方法:以耐药性白血病K562/ADM细胞为靶细胞,采用MTT比色法测定细胞增殖活性;细胞形态学和AnnexinV/PI双染色法检测细胞凋亡,电镜观察凋亡细胞内质网形态结构变化;实时定量RT-PCR检测GRP78mRNA的表达;Western blot法检测GRP78蛋白的表达。结果:不同浓度的辣椒素显著抑制K562/ADM细胞的增殖活性,20、50μmol/L辣椒素诱导后K562/ADM细胞出现典型的凋亡形态学改变,细胞凋亡率明显增高,分别为39.67%和41.78%。辣椒素诱导凋亡过程中,K562/ADM细胞出现内质网明显扩张和脱颗粒现象,GRP78mRNA的表达增高,GRP78蛋白的表达量分别增高1.5和2.2倍,随时间延长有所降低。结论:辣椒素可能通过内质网应激反应性途径诱导K562/ADM细胞发生凋亡。
AIM：To investigate whether capsaicin induces apoptosis in drug-resistant leukemia K562/ADM cells mediated by endoplasmic reticulum stress-related pathway.METHODS：The drug-resistant leukemia K562/ADM cells were used as the target cells.The cell proliferating activity was assessed with MTT colorimetric assay.The apoptosis of K562/ADM cells was identified by double staining of FITC-Annexin V and propidium iodide（PI）and the cellular morphology.The ultrastructure of the cells and endoplasmic reticulum were observed by transmission electron microscopy.The expression of GRP78 gene mRNA and protein were analyzed by real-time reverse transcription-polymerase chain reaction（RT-PCR）and Western blotting,respectively.RESULTS：Capsaicin of different concentrations markedly inhibited the proliferation of K562/ADM cells and also induced the cells to apoptosis.After the treatment with 20 and 50 μmol/L capsaicin for 48 h,K562/ADM cells showed the typical morphological characteristics of apoptosis,and the apoptotic rate of the cells increased to 39.67% and 41.78%,respectively.During the apoptotic process of K562/ADM cells induced with capsaicin,the endoplasmic reticulum exhibited obvious expansion and degranulation.The expression of GRP78 gene mRNA of K562/ADM cells increased and then decreased slightly,the expression of GRP78 protein increased by 1.5 and 2.2 times respectively,and gradually reduced with the inducing time.CONCLUSION：Capsaicin may induce drug-resistant K562/ADM cells to apoptosis via endoplasmic reticulum stress-mediated pathway.
出处
《中国临床药理学与治疗学》
CAS
CSCD
2010年第8期856-860,共5页
Chinese Journal of Clinical Pharmacology and Therapeutics
基金
甘肃省科技支撑计划项目(1011FKCA145)
甘肃省新药临床前研究重点实验室(GSKFKT-0701)
甘肃省天然药物重点实验室(0804)开放基金资助项目
作者简介
侯娟娟，女，硕士研究生，研究方向：生物化学与分子生物学。Tel：09318915391E—mail：houjj08@lzu．cn
魏虎来，通信作者，男，教授，博士研究生导师，研究方向：细胞分子生物学。Tel：09318915082E-mail：weihulai@lzu．edu．cn
