首页  期刊介绍 征稿简则 编委会 期刊征订 广告服务 留言板 联系我们 English

中国应用生理学杂志 ›› 2019, Vol. 35 ›› Issue (4): 312-316.doi: 10.12047/j.cjap.5799.2019.066

• 研究论文 • 上一篇    下一篇

2-12烷基-6-甲氧基环己基-2,5-二烯-1,4-二酮(DMDD)抗弥漫大B淋巴瘤的作用及机制*

洪凯+, 蒋盼若+, 柯瑞君, 应佳豪+, 张肖艳+, 陈佳玉   

  1. 绍兴文理学院医学院, 浙江 绍兴 312000
  • 收稿日期:2018-12-24 出版日期:2019-07-28 发布日期:2019-11-06
  • 通讯作者: , Tel: 0575-88345826; E-mail: chenkc1969@163.com.
  • 基金资助:
    *国家自然科学基金资助项目(81402979);浙江省科技厅资助项目(2015C33270);浙江省卫生厅资助项目(2014KYA230);国家大学生创新创业训练计划项目(201810349028);浙江省大学生科技创新活动计划项目(2018R432016);绍兴市大学生创新项目(SXSDC201709)

Effects of 2-12alkyl-6-methoxycyclohexa-2, 5-diene-1, 4-dione(DMDD)on diffuse large B lymphoma and its mechanism

HONG Kai+, JIANG Pan-ruo+, KE Rui-jun, YING Jia-hao+, ZHANG Xiao-yan+, CHEN Jia-yu   

  1. Medical School, Shaoxing University, Shaoxing 312000, China
  • Received:2018-12-24 Online:2019-07-28 Published:2019-11-06

摘要: 目的:探讨2-12烷基-6-甲氧基环己基-2,5-二烯-1,4-二酮(DMDD)抗弥漫大B淋巴瘤(DLBCL)的作用及分子机制。方法:动物实验取4周龄BALB/C小鼠,分5组,20只/组,腹股沟注射DLBCL细胞株OCI-LY19细胞1 × 107 cells/ml 每只0.1 ml,两天后分别灌胃0、1、5、25、125 mg/kg剂量的DMDD,1次/2天,给药的第18日,杀10只小鼠,取瘤组织称重,记录剩余小鼠的生存期。细胞实验取OCI-LY19细胞加入96孔培养板,每孔100 μl 1×105 cells/ml,分别加入100 μl DMDD使其终浓度分别为0、1、5、25和125 μmol/L,作用0、24、48和72 h,设三复孔,MTS法检测细胞增殖活性;根据细胞增殖实验结果,选择0 μmol/L、5 μmol/L和25 μmol/L的DMDD作为后续用药浓度作用OCI-LY19细胞24 h,流式细胞仪分析凋亡率,hoechst染色观察细胞核型,JC-1染色观察线粒体膜电位,LDH释放实验评估药物细胞毒性,qPCR、Western blot分析基因转录和表达水平。结果:动物实验表明:与0 mg/kg用药组比,1~125 mg/kg DMDD能抑制小鼠瘤组织生长并延长其生存期(P<0.01)。细胞实验表明:DMDD用药组OCI-LY19细胞增殖活性明显降低、凋亡水平显著增加(P<0.01),细胞核出现碎裂、凝集和凋亡小体及线粒体膜电位下降,LDH释放率显著增加(P<0.01),细胞内caspase-3和bax基因的转录表达和IκBα的磷酸化水平显著上调,bcl-2、bcl-xL、jak2和stat3基因的转录和蛋白表达水平明显受抑(P<0.01)。结论:DMDD通过抑制JAK2/STAT3和NF-κB信号通路中JAK2、STAT3和p-IκBα的表达,下调BCL-2/BAX、活化Caspase-3,最终激活OCI-LY19细胞线粒体凋亡的内源性通路而促进了DLBCL细胞凋亡,抑制了OCI-LY19细胞增殖,具有抗DLBCL的作用。

关键词: 2-12烷基-6-甲氧基环己基-2,5-二烯-1,4-二酮, 弥漫大B细胞淋巴瘤, 细胞凋亡, 信号通路, 小鼠, 生存期(时间)

Abstract: Objective: To investigate the effects and molecular mechanisms of 2-12alkyl-6-methoxycyclohexa-2,5-diene-1,4-dione(DMDD) on diffuse large B lymphoma (DLBCL).Methods: In animal experiments, 4-week-aged BALB/C mice were divided into 5 groups, 20 mice in each group. Mice were inguinal injected with DLBCL cell line OCI-LY19 cells 0.1 ml at the concention of 1 × 107 /ml. Two days later, mice were treated with DMDD at the doses of 0, 1, 5, 25 and 125 mg/kg by intragastric administration respectively, once /2 days. Ten mice of each group were killed on the 18th day of administration, and the tumor tissues were weighed. The survival time of the remaining mice were recorded. In cell experiments, OCI-LY19 cells were added to 96-well culture plates, 100 μl 1×105 cells/ml per well, then 100 μl DMDD was added to the well and the final concentrations were 0, 1, 5, 25 and 125 μmol/L respectively. The cells were treated with DMDD for 0, 24, 48 and 72 h, three wells in each group. The cell proliferation activity was detected by MTS assay. According to the results of cell proliferation experiments, OCI-LY19 cells were treated with DMDD at the concentrations of 0 μmol/L, 5 μmol/L and 25 μmol/L for 24 h. The apoptosis rate was analyzed by flow cytometry, the nuclear type was observed by hoechst staining, the mitochondrial membrane potential was observed by JC-1 staining, cytotoxicity of drugs was evaluated by LDH release experiment, gene expression and transcription were analyzed by qPCR and Western blot.Results: Compared with 0 mg/kg drug group, DMDD at the dose of 1~125 mg/kg could inhibit the growth of tumor tissue in mice and prolong their survival time (P<0.01). Cell experiments showed: in DMDD group, the proliferation activity of OCI-LY19 cells was decreased significantly and the level of apoptosis was increased significantly (P<0.01), nuclear fragmentation, agglutination, apoptotic bodies occurred and mitochondrial membrane potential was decreased, the LDH release rate was increased significantly (P<0.01), the expressions of caspase-3 and bax genes and the phosphorylation level of Ikappa B alpha in cells were up-regulated significantly, the protein expression levels of bcl-2, bcl-xL, jak2 and stat3 were inhibited significantly (P<0.01).Conclusion: DMDD can inhibit the expressions of JAK2, STAT3 and p-Ikappa B alpha in JAK2/STAT3 and NF-kappa B signal pathways, down-regulate BCL-2/BAX and activate Caspase-3, finally, activate the endogenous pathway of mitochondrial apoptosis in OCI-LY19 cells and promote the apoptosis of DLBCL cells, inhibit proliferation of OCI-LY19 cells. It has inhibitive effects on DLBCL.

Key words: DMDD, diffuse large B lymphoma (DLBCL), cell apoptosis, signal pathway, mice, survival time

版权所有 © 2015 《中国应用生理学杂志》编辑部
京ICP备16058274号-1
地址:天津市和平区大理道1号,邮编:300050  电话:022-84655184  E-mail:tjzgyish@163.com
本系统由北京玛格泰克科技发展有限公司设计开发 技术支持:support@magtech.com.cn