摘要
[目的]探讨苦木(PQ)提取物对耐药性肺癌细胞的促凋亡作用及其机制.[方法]用不同质量浓度PQ提取物(0,0.02,0.04,0.06,0.08,0.10 g/L)处理耐药性肺癌细胞24 h,通过MTT Assay、荧光显微镜照相、划痕实验及集落形成实验检测细胞毒性、细胞存活率、细胞内活性氧水平及细胞凋亡情况;PQ提取物(0.08 g/L)处理细胞不同时间(0,1,3,6,12,24 h),采用Western Blot检测凋亡相关蛋白表达情况.[结果]PQ提取物(0,0.02,0.04,0.06,0.08,0.10 g/L)处理细胞24 h可有效降低A549/PTX细胞增殖和迁移能力,抑制A549/PTX细胞活力(P<0.05,P<0.001);荧光显微照相观察结果显示,PQ提取物可增加细胞内线粒体活性氧聚积;Western Blot检测结果显示,PQ提取物处理后引发细胞自噬,同时时间依赖性地诱导线粒体依赖性细胞凋亡.[结论]PQ提取物通过升高细胞内活性氧、损伤线粒体膜电位、降低细胞增殖及迁移能力及发生自噬最终导致耐药性肺癌细胞发生凋亡.
OBJECTIVE To investigate the pro-apoptotic effects of Picrasma Quassioides(PQ)extracts on drug-resistant lung cancer cells.METHODS The paclitaxel resistant lung cancer cells were treated with the different concentrations of 0,0.02,0.04,0.06,0.08 and 0.10 g/L PQ extracts for incubation 24 h,then the cellular cytotoxicity,survival rate,intracellular reactive oxygen species levels,cellular apoptosis and apoptosis-related protein expression levels were determined by MTT Assay,fluorescence microscopy,scratch test,colony formation assay,the Western Blot analysis was used to detect apoptosis related proteins in the cells treated with 0.08 g/L extracts of PQ at the same time in a time-dependent manner(0,1,3,6,12,24 h).RESULTS The extracts of PQ(0,0.02,0.04,0.06,0.08,0.10 g/L)incubation cells for 24 h,the PQ could effectively inhibit the proliferation and migration of A549/PTX cells and reduce the activity of A549/PTX cells as well as induce the cellular autophagy(P<0.05,P<0.001).The results of fluorescence microscopy showed that extracts of PQ could increase the accumulation of reactive oxygen species in mitochondria;Western Blot showed that extracts of PQ could induce autophagy and induce mitochondrial dependent apoptosis in a time-dependent manner.All of these phenomenons due to increasing the accumulation of mitochondrial reactive oxygen species which in turn induces the mitochondria dependent apoptosis signaling pathway activation.CONCLUSION PQ extracts can induce apoptosis of drug-resistant lung cancer cells by increasing intracellular reactive oxygen species,damaging mitochondrial membrane potential,reducing cell proliferation and migration,and autophagy.
作者
金永哲
谢丹萍
孙虎男
JIN Yongzhe;XIE Danping;SUN Hunan(Yanbian University College of Nursing,Yanji 133002,Jilin,China;College of Life Science&Technology,Heilongjiang Bayi Agricultural University,Daqing 163319,Heilongjiang,China)
出处
《延边大学医学学报》
CAS
2020年第4期243-250,共8页
Journal of Medical Science Yanbian University
关键词
苦木
紫杉醇耐药肺癌细胞
活性氧
细胞凋亡
Picrasma Quassioides
paclitaxel resistant lung cancer cells
reactive oxygen species
apoptosis
作者简介
金永哲(1969—),男(朝鲜族),副教授,硕士,研究方向为抗癌药物筛选及活性评价;通信作者:孙虎男,Email:sunhunan76@163.com.
