冬凌草甲素对耐顺铂鼻咽癌细胞株HNE1/DDP凋亡的影响  

Effects of Oridonin on Apoptosis of Cisplatin-Resistant Nasopharyngeal Carcinoma HNE1/DDP Cells

作　　者：鲁星月 时宗芬 陈长江 朱晨露 张配 刘浩 赵素容 Lu Xingyue;Shi Zongfen;Chen Changjiang;Zhu Chenlu;Zhang Pei;Liu Hao;Zhao Surong(School of Pharmacy,Bengbu Medical College/Anhui Provincial Engineering Technology Research Center of Biochemical Pharmaceuticals,Anhui Bengbu 233030)

机构地区：[1]蚌埠医学院药学院/安徽省生化药物工程技术研究中心,蚌埠233030

出　　处：《中药药理与临床》2020年第1期68-73,共6页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金项目(81603155);安徽省自然科学基金青年基金项目(1708085QH212);安徽省高等学校自然科学研究重点项目(KJ2016A486);安徽省大学生创新创业训练项目(201710367061)。

摘　　要：目的:探讨冬凌草甲素对耐顺铂鼻咽癌细胞株HNE1/DDP凋亡的影响。方法:MTT实验检测冬凌草甲素对HNE1/DDP细胞增殖能力的影响;集落克隆实验检测细胞集落形成能力;DAPI荧光染色法检测HNE1/DDP细胞核形态的变化;PI单染流式细胞术分析细胞的凋亡情况;检测试剂盒测定细胞内ROS的水平;Western blot检测细胞凋亡相关蛋白表达水平。结果:MTT实验结果表明,冬凌草甲素20μmol/L^50μmol/L可显著减弱HNE1/DDP细胞活力,抑制细胞增殖;相比于模型对照组,3μmol/L^5μmol/L的冬凌草甲素对HNE1/DDP细胞的集落形成能力有明显的抑制作用(P <0. 05或P <0. 01);且随着冬凌草甲素给药浓度的增加,细胞核发生明显皱缩,核碎裂增加;细胞内ROS水平明显增加;抑凋亡蛋白Mcl-1、Bcl-2表达明显下调,促凋亡蛋白Bax、Bak表达明显上调(P <0. 05或P <0. 01)。结论:冬凌草甲素能显著抑制耐顺铂鼻咽癌HNE1/DDP细胞的增殖,并诱导其发生凋亡,其作用可能是通过诱导细胞内ROS的产生,对Mcl-1、Bcl-2、Bax、Bak等凋亡相关蛋白进行调控实现的。Objective: To investigate the effect of oridonin on apoptosis of cisplatin-resistant nasopharyngeal carcinoma HNE1/DDP cells. Methods: MTT assay was used to detect the inhibitory effect of oridonin on the proliferation of HNE1/DDP cells. Colony formation assay was usedto detect the forming ability of HNE1/DDP cells. Nuclear morphological changes were detected by DAPI fluorescence staining assay. The apoptosis of cells was analyzed by flow cytometry with PI staining assay. Reactive oxygen species( ROS) detection kit was applied to detect the level of intracellular ROS. The expression of apoptosis-related protein in HNE1/DDP cells was detected by Western blot. Results: MTT assay showed that oridonin significantly reduced the cell viability and inhibited the proliferation of HNE1/DDP cells. Compared with the control group,3 μmol/L ~ 5 μmol/L oridonin significantly inhibited the colony formation ability of HNE1/DDP cells. With the increase of concentration of oridonin,the nucleus of the cells shrank and the nucleus fragmentation was increased,the intracellular ROS level was obviously enhanced,the expression of anti-apoptotic protein Mcl-1 and Bcl-2 was down-regulated and the expression of pro-apoptotic protein Bax and Bak was up-regulated. Conclusion: Oridonin can significantly inhibit the proliferation of cisplatin-resistant nasopharyngeal carcinoma HNE1/DDP cells and induce apoptosis,which may be related to inducing intracellular ROS production and regulating the expression levels of Mcl-1,Bcl-2,Bax and Bak.

关 键 词：冬凌草甲素 鼻咽癌 细胞增殖 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]