摘要
目的探讨紫花牡荆素对肺癌A549细胞（简称肺癌细胞）增殖及凋亡的影响,为其用于临床治疗肺癌提供依据。方法取对数生长期的肺癌细胞,分别加入终浓度为0.1、0.5、1、5、10、25、50、100μmol/L紫花牡荆素溶液,分别作用24、48 h,采用SRB法检测增殖抑制率,计算50%抑制浓度（IC50）。取对数生长期的肺癌细胞,分别加入最终浓度为0、5、15μmol/L紫花牡荆素溶液,Hoechst33258染色后共聚焦显微镜下观察细胞凋亡形态;采用流式细胞术检测细胞周期时相分布。结果随紫花牡荆素浓度升高,作用24、48 h的肺癌细胞增殖抑制率均呈上升趋势。经0.5～100μmol/L紫花牡荆素作用48 h的肺癌细胞增殖抑制率均明显高于同一浓度作用24 h的肺癌细胞（P均〈0.05）。紫花牡荆素作用24、48 h时的IC50分别为14.74、8.16μmol/L。随紫花牡荆素浓度升高,作用24 h的肺癌细胞核染色质固缩偏向一边、核裂解和细胞碎片等细胞凋亡形态学改变越来越明显;与同浓度紫花牡荆素作用24 h比较,作用48 h的肺癌细胞凋亡形态学改变更明显。经0、5、15μmol/L紫花牡荆素作用24、48 h的肺癌细胞凋亡率均逐渐升高,处于G1、S期的比例均逐渐降低,处于G2/M期的比例均逐渐升高（P均〈0.05）。与同一浓度紫花牡荆素作用24 h的肺癌细胞比较,经5、15μmol/L紫花牡荆素作用48 h的肺癌细胞凋亡率均升高,处于G1、S期的比例均降低,处于G2/M期的比例均升高（P均〈0.05）。结论紫花牡荆素可将肺癌细胞阻滞于G2/M期,具有增殖抑制作用和凋亡促进作用,并呈时间、浓度依赖性。
Objective To investigate the influence of casticin on the proliferation and apoptosis of lung cancer A549cells（ lung cancer cells for short） and to provide basis for the clinical treatment of lung cancer. Methods We chose the lung cancer cells in the logarithmic phase and separately treated them with different concentrations of casticin（ 0. 1,0. 5,1,5,10,25,50 and 100 μmol / L）. The SRB method was used to detect the anti-proliferative rate of A549 cells in order to finally calculate its half inhibitory concentration（ IC50）. Furthermore,A549 lung cancer cells in the logarithmic phase were treated with final concentrations of casticin（ 0,5 and 15 μmol / L） and meanwhile we observed their morphology of apoptosis by laser scanning confocal microscope after being treated with Hoechst33258 staining. The apoptosis rate of A549 cells was detected by flow cytometry. Results The anti-proliferative rates of A549 cells at 24 and 48 h were significantly increased with the increasing concentrations of casticin. In addition,after being treated with casticin from 0. 5 to 100μmol /L,the anti-proliferative rates of A549 cells at 48 h were significantly higher than those at 24 h（ all P〈0. 05）. The IC50 of casticin were separately 14. 74 μmol / L at 24 h and 8. 16 μmol / L at 48 h. With the increasing concentrations of casticin,cell morphology at 24 h showed obvious changes,such as cell shrinkage,chromatin condensation,nucleus fragment and cellular debris. The morphology changes of A549 cells at 48 h were more obviously than those at 24 h. The apoptosis rates of A549 cells which were separately treated by casticin with concentrations of 0,5 and 15 μmol / L for 24 and 48 h were all gradually increased. Furthermore,the proportion of cells at G1 and S periods were all decreased and cells at G2/ M period were increased（ all P〈0. 05）. The apoptosis rates of A549 cells treated by 5 and 15 μmol / L casticin at 48 h wereall increased as compared to those at 24 h. With the same comparison,the proportion of cells at G1 and S periods were both decreased and the cells at G2/ M were increased（ all P〈0. 05）. Conclusion Casticin may block the cell cycle of A549 cells at G2/ M period and is capable of anti-proliferation as well as promoting apoptosis,which is in a time- and dose-dependent manner.
出处
《山东医药》
CAS
北大核心
2016年第36期13-16,I0002,共5页
Shandong Medical Journal
基金
新疆生产建设兵团社会发展科技攻关与成果转化计划项目(2015AD007)
国家级大学生创新创业训练计划项目(201510759066)
作者简介
张春春（1988-），女，硕士研究生，研究方向为药物新制剂与新剂型研究。E—mail：1271403480@qq．com
通信作者简介：应雪（1981-），女，博士、副教授，研究方向为药物新剂型研究。E-mail：yingxue2011@qq．com
