自噬在As2O3诱导Burkitt淋巴瘤Raji细胞死亡中的作用及机制研究  

Role and mechanism of autophagy in the arsenic trioxide-induced death of Burkitt lymphoma Raji cells

作　　者：李彩丽[1,2] 陈静[1] 王蓓[1] 王菲菲[1] 田宝莹 谢蓓[1] 范临兰[1] 魏虎来[1] 

机构地区：[1]兰州大学基础医学院 甘肃省新药临床前研究重点实验室,甘肃兰州730000 [2]西北民族大学医学院生物化学与分子生物学教研室,甘肃兰州730030

出　　处：《中国药理学通报》2014年第5期719-724,共6页Chinese Pharmacological Bulletin

基　　金：西北民族大学中央高校基本科研业务费专项资助项目(No zyz2011094);甘肃省兰州市城关区科技局科技计划项目(No 2013-7-17)

摘　　要：目的研究自噬在As2O3诱导的Raji细胞死亡中的作用及机制。方法透射电镜和MDC荧光染色观察细胞自噬形态,MTT法测定细胞增殖活性;流式细胞术(FCM)及FITC-Annexin-V/PI染色检测细胞周期和细胞凋亡,Western blot检测细胞LC3-Ⅰ/Ⅱ表达及转换;RT-PCR检测细胞bcl-2 mRNA和p53 mRNA的表达水平。结果 As2O3明显抑制Raji细胞增殖,诱导其细胞周期G2/M阻滞和凋亡,抑制bcl-2基因和增强p53基因表达。同时,As2O3可同步诱发Raji细胞的自噬活性。自噬抑制剂3-甲基腺嘌呤(3-MA)抑制As2O3诱导的自噬活性,上调bcl-2基因和下调p53基因表达,抑制As2O3对Raji细胞的杀伤效应、降低Raji细胞对As2O3的敏感性。而自噬诱导剂雷帕霉素(Rapa)的作用则与3-MA的效应正好相反。结论细胞凋亡和自噬性细胞死亡共存于As2O3诱导的Raji淋巴瘤细胞死亡中,Bcl-2和p53可能起着关键的调控作用。Aim To investigate the role of autophagy and its mechanism in Raji cell death induced by arsenic trioxide. Methods Transmission electron microscopy （SEM） and MDC fluorescence staining were used to observe autophagy. MTT colorimetry was employed to assay the cellular proliferating activity. Cell apoptosis and cell cycle analysis were performed using FITC- Annexin-V/PI double staining and flow cytometry （FCM）. The expressions of LC3 and the conversion of LC3-I to LC3-II were measured by western bloting. The expression of bcl-2 mRNA and p53 mRNA were detected by reverse transcription-polymerase chain reaction （RT-PCR）. Results Arsenic trioxide could obviously inhibit the proliferation of Raji cells, arrest the cells at G2/M phase and induce apoptosis. Meanwhile, arsenic trioxide markedly inhibited the expression of bcl-2 mRNA and enhanced the expression of p53 mRNA in Raji cells. Arsenic trioxide also induced autophagy synchronously which paralleled with the induction of apoptosis in Raji cells, and 3-MA, an autophagy inhibitor, was able to reverse the arsenic trioxide-activated autophagic activity, up-regulate bcl-2,down-regulated p53 expression and suppress the lethal effect of arsenic trioxide on sensitivity to arsenic trioxide. Raji cells to reduce their In contrast, the Rapamycin, an autophagy inducer, possessed the completely opposite effects on Raji cells compared with 3-MA.Conclusions The apoptosis and autophagic cell death are coexistent in arsenic trioxide-triggered death of Raji lymphoma cells, and Bcl-2 and p53 may play a key regulating role in this process.

关 键 词：自噬 三氧化二砷 RAJI细胞 细胞凋亡 Bcl-2 p53 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]