As_2O_3介导弥漫大B细胞淋巴瘤细胞凋亡的JNK磷酸化机制研究

The Involvement of JNK Phosphorylation in Diffuse Large-B Cell Lymphoma Apoptosis Mediated by As_2O_3

目的探讨JNK磷酸化在As2O3诱导的弥漫大B细胞淋巴瘤细胞凋亡过程中的参与机制。方法人弥漫大B细胞淋巴瘤细胞株LY1随机分为5组:对照组、三氧化二砷低剂量、中剂量、高剂量组及JNK阻断剂SP600125组。对照组和三氧化二砷组细胞给予DMSO或三氧化二砷(1Μm,5μM和25μM)孵育20 h。SP600125组在三氧化二砷(25μM)孵育的同时加入SP600125(10μM)孵育。CCK-8测定各组LY1细胞活力情况,q PCR和Western Blot分析各组细胞中细胞凋亡蛋白Bax、Bcl2、Caspase9及JNK和p-JNK蛋白的表达变化情况。结果人弥漫大B细胞淋巴瘤细胞株LY1随着三氧化二砷作用浓度升高而活力逐渐降低,且各组之间具有显著的统计学差异性(P<0.01)。q PCR和Western Blot结果显示三氧化二砷各组细胞凋亡蛋白Bax和Caspase9表达升高,而Bcl2表达降低。同时,三氧化二砷组中p-JNK1/2表达明显增高。而SP600125组上述异常均得到部分恢复。结论 JNK磷酸化过程参与了三氧化二砷诱导的弥漫大B细胞淋巴瘤的细胞凋亡过程。

Objective To explore the participation of JNK phosphorylation in the apoptosis of diffuse large-B cell lymphoma induced by As2O3. Methods Human DLBCL cell line LY1 was divided into the control group,As2O3 group and JNK inhibitor SP600125 group. And As2O3 group were divided into the low dose,middle dose and high dose group. The control group and As2O3 group were incubated with DMSO or As2O3( 1 Μm,5 μM and 25 μM) for 20 h. SP600125 cells were incubated with As2O3( 25 μM) and SP600125( 10 μM). The cellular viability was calculated by CCK-8 and apoptotic proteins Bax、Bcl2、Caspase9,JNK and p-JNK were detected by q PCR and Western Blot. Results The viability of LY1 decreased with the increase of As2O3 concentration,there had significant difference( P < 0. 01). q PCR and Western Blot showed that apoptotic proteins Bax and Caspase9 increased in As2O3 group while Bcl2 decreased. p-JNK in As2O3 group increased dramatically. SP600125 can alleviate the aforementioned parameters greatly. Conclusion The apoptosis of LY1 cell line induced by As2O3 is regulated by apoptotic proteins that mediated by JNK phosphorylation.