小白菊内酯对人小细胞肺癌NCI-H446细胞增殖及凋亡作用研究  

Effects of parthenolide on the proliferation and apoptosis of small cell lung cancer cell line NCI-H446

作　　者：刘捷[1] 徐玲[1] Liu Jie;Xu Ling(Department of Respiratory, The First Affiliated Hospital of Chongqing Medical Universit)

机构地区：[1]重庆医科大学附属第一医院呼吸内科,重庆400016

出　　处：《重庆医科大学学报》2018年第3期414-419,共6页Journal of Chongqing Medical University

基　　金：重庆市卫生和计生委中医药科技资助项目(编号:zy201402106)

摘　　要：目的:研究小白菊内酯(parthenolide,PTL)对人小细胞肺癌(small cell lung cancer,SCLC)细胞株NCI-H446增殖及凋亡作用的影响及其可能的分子机制。方法:NCI-H446细胞株由武汉大学细胞库提供,PTL购于Gene Operation公司,将NCIH446细胞株分为对照组和实验组(1.5μg/m L组、2.0μg/m L组、2.5μg/m L组和3.0μg/m L组),向实验组细胞培养基中加入对应浓度PTL处理细胞,对照组中加入等体积无血清培养基。MTT比色法观测不同浓度PTL对NCI-H446细胞增殖的影响;Hochest 33258染色后,荧光显微镜下观察细胞核形态学变化;流式细胞术(flow cytometry,FCM)分析细胞凋亡率;RT-PCR分别检测不同浓度组PTL处理48 h后细胞中NF-κB、Caspase-3、Caspase-8、Caspase-9基因的表达状态,Wsetern blot检测NF-κB、Caspase-3、Caspase-8、Caspase-9蛋白的表达状态。结果:PTL抑制小细胞肺癌NCI-H446细胞增殖并呈浓度依赖性;Hochest33258染色结果显示,PTL作用NCI-H446细胞48 h后,细胞数量减少,细胞核固缩碎裂,形成凋亡小体,导致细胞凋亡;流式细胞术检测显示,PTL处理NCI-H446细胞后,细胞凋亡率与药物浓度梯度呈浓度依赖性,差异有统计学意义(P<0.05);RT-PCR及Wsetern blot结果显示,与对照组比较,经PTL处理的NCI-H446细胞中NF-κB m RNA及蛋白表达下调,同时Caspase-3、Caspase-8、Caspase-9 m RNA及蛋白表达上调,差异具有统计学意义(P<0.05)。结论:PTL能抑制NCI-H446细胞的增殖并诱导其发生凋亡,PTL的作用机制可能与抑制NF-κB信号通路及活化Caspase系统有关。Objective：To investigate the effects of parthenotide （PTL） on the proliferation and apoptpsis of human proliferation and apoptosis of human small cell lung cancer cell line NCI-H446 and its mechanisms. Methods ：Cell line provided by the cell library of Wuhan University. PTL was bought in company of gene operation. NCI-H446 ceils were divided into control group and treatment group（ 1.5 μg/mL, 2.0μg/mL, 2.5μg/mL, 3.0μg/mL）. Treatment group were treated with different concentrations of PTL by adding corresponding concentration of PTL into culture medium,while control group was added equal volume culture medium without fetal calf serum. The cellular proliferative activity was determined by MTlr assay. Morphologic changes were observed by fluorescence microscope. The apoptosis rates of cell were examined by flow eytometry. The expression of NF-KB, Caspase-3, Caspase-8 and Caspase- 9 genes of NCI-H446 cells was detected by RT-PCR assay. The expression of NF-KB, Caspase-3, Caspase-8 and Caspase-9 proteins of NCI-H446 ceils was evaluated by Western blot. Results ：Parthenolide inhibited the proliferation of NCI-H446 cells in a dose-de- pendent manner. Fluorescent microscopy showed that the number of NCI-H446 cells was reduced under Hoechst 33258 staining;the nucleus pyenosis fracture and apoptotic bodies were observed. Flow eytometry showed that the cell apoptosis rate was positively cor- related with the concentration of PTL,with statistically significant differences（P〈0.05）. Compared with those of control group, RT- PCR and Western blot revealed that the expressions of mRNA and protein of NF-KB in NCI-H446 cells were reduced,at the same time, the expressions of Caspase-3 ,-8 ,-9 mRNA and proteins were increased with statistically significant differences（P〈0.05 ）.Conclusion：PTL could repress cellular proliferation activity and induce apoptosis of NCI-H446 ceils. The mechanisms could relate to inhibition of NF-KB signaling pathway and activation of caspases system.

关 键 词：小白菊内酯 小细胞肺癌HCI-H446 细胞增殖 细胞凋亡 

分 类 号：R347[医药卫生—基础医学]