三氧化二砷诱导套细胞淋巴瘤细胞株凋亡及机制研究

Effect of Arsenic Trioxide on Induction of Apoptosis in MCL Cell Line and Its Possible Mechanisms

本研究旨在探讨三氧化二砷(ATO)诱导套细胞淋巴瘤(MCL)细胞株凋亡的效应及其机制。以不同浓度的ATO处理细胞,再通过MTT法检测细胞MCL细胞株(jeko-1,mino,JVM-2)增殖;用AnnexinⅤ-FITC/PI双染流式细胞术检测jeko-1细胞的凋亡;用DiOC6(3)染色流式细胞术检测jeko-1细胞的线粒体跨膜电位丢失;用Western blot检测细胞周期蛋白D1(cyclin D1)及凋亡相关蛋白MCL-1,BCL-2,PUMA,NOXA,cCaspase-3,cCaspase-9,cPARP在ATO处理前后的表达变化。结果表明:ATO抑制MCL细胞增殖,诱导MCL细胞凋亡,并且引起MCL细胞线粒体跨膜电位丢失,引起MCL-1,PUMA,cyclin D1蛋白表达水平降低,cPARP,cCaspase-3,cCaspase-9表达水平增加,不影响BCL-2,NOXA表达。结论:ATO能有效抑制MCL细胞增殖,诱导MCL细胞株凋亡,其中细胞凋亡的线粒体途径起着重要的作用。

This study was aimed to explore the effect of arsenic trioxide （ATO） on proliferation and apoptosis of mantle cell lymphoma（MCL） cell lines and the underling mechanisms of the apoptosis. MCL cell lines （jeko-1, mino, JVM-2） were treated with different concentrations of ATO, then growth profile of these cells were detected by MTT. Apoptosis of ATO-treated jeko-1 cells were detected by flow cytometry with Annexin V-FITC/PI double staining. The loss of mitochondrial membrane potential of ATO-treated jeko-1 cells were detected by FCM with DiOC6 （ 3 ） staining. The expressions of cyclin D1 and apoptosis related proteins MCL-1, BCL-2, PUMA, NOXA, cCaspase-3 （cleaved caspase-3 ） ,cCaspase-9 （cleaved caspase-9）, cPARP （cleaved PARP） were detected by Western blot. The results indicated that ATO inhibited cell growth, induced apoptosis of MCL cells and disrupted mitochondrial membrane potential. ATO could decrease expressions of MCL-1, PUMA and cyclin D1, increase expressions of cPARP, cCaspase-3, cCaspase-9 and the expressions of BLC-2 and NOXA were not changed. It is concluded that ATO can induce cell growth arrest and apoptosis of MCL ceils. The mitochondrial pathway plays a very important role in cell apoptosis.