冬凌草乙素联合放疗对肺癌A549细胞凋亡的协同作用及miR-16-5p/WEE1信号轴的影响  

Synergistic Effect of Ponicidin Combined with Radiotherapy on the Apoptosis of Lung Cancer A549 Cells and Its Influence on MiR-16-5p/WEE1 Signal Axis

作　　者：王朔[1] 翟士媛 王昆 李鹏 纪春梅[1] Wang Shuo;Zhai Shiyuan;Wang Kun;Li Peng;Ji Chunmei(Department of Respiratory Medicine,Tangshan Union Medical College Hospital,Hebei Tangshan 063000,China;Department of Critical Medicine,Affiliated Hospital of North China University of Technology)

机构地区：[1]唐山市协和医院呼吸内科,河北唐山063000 [2]华北理工大学附属医院重症医学科

出　　处：《中国药师》2021年第5期824-828,共5页China Pharmacist

基　　金：河北省医学科学研究重点课题计划项目(编号:20181300)。

摘　　要：目的:探讨冬凌草乙素联合放疗对肺癌A549细胞凋亡的协同作用及微小RNA-16-5p(miR-16-5p)/丝氨酸/苏氨酸蛋白激酶1(WEE1)信号轴的影响。方法:设A549细胞组、放疗组(6 Gy·min^(-1)的固定剂量率发射,照射孵育24 h)、冬凌草乙素组(20 mg·L^(-1))、放疗+冬凌草乙素组(冬凌草乙素浓度为20 mg·L^(-1),并以6 Gy·min^(-1)的固定剂量率发射,照射孵育24 h);培养结束后,CCK-8法测定细胞增殖水平,Transwell腔室系统测定细胞侵袭水平,流式细胞仪测定细胞凋亡水平,实时荧光逆转录聚合酶链反应(RT-PCR)及蛋白免疫印迹法测定细胞miR-16-5p、WEE1 mRNA和蛋白水平。结果:与A549细胞组比较,放疗组、冬凌草乙素组、放疗+冬凌草乙素组光密度(OD)值、存活率、穿膜数、WEE1 mRNA和蛋白表达显著降低(P<0.05),凋亡率、miR-16-5p表达显著升高(P<0.05);与放疗组、冬凌草乙素组比较,放疗+冬凌草乙素组OD值、存活率、穿膜数、WEE1 mRNA和蛋白表达显著降低(P<0.05),凋亡率、miR-16-5p表达显著升高(P<0.05)。结论:冬凌草乙素联合放疗对肺癌A549细胞凋亡具有协同作用,其机制可能与冬凌草乙素促进miR-16-5p的表达进而抑制WEE1表达有关。Objective:To investigate the synergistic effect of ponicidin combined with radiotherapy on the apoptosis of lung cancer A549 cells and the effect on micro RNA-16-5p(miR-16-5p)/WEE1 signal axis.Methods:A549 cell group,radiotherapy group(6 Gy·min^(-1)fixed dose rate radiation,incubation for 24 hours),ponicidin group(20 mg·L^(-1))and radiotherapy+ponicidin group(ponicidin concentration of 20 mg·L^(-1),and 6 Gy·min^(-1))were set up.At the end of culture,cell proliferation was measured by CCK-8 method,invasion level was measured by Transwell chamber system,apoptosis level was measured by flow cytometry,and the levels of miR-16-5p and WEE1 mRNA and protein were determined by real-time fluorescent reverse transcription(RT-PCR)and Western blot.Results:Compared with those in A549 cell group,the OD value,survival rate,penetration number,WEE1 mRNA and protein expressions in radiotherapy group,ponicidin group and radiotherapy+ponicidin group were decreased(P<0.05),while the apoptosis rate and miR-16-5p expression were increased(P<0.05).Compared with those in radiotherapy group and ponicidin group,the OD value,survival rate,penetration number,WEE1 mRNA and protein expressions in radiotherapy+ponicidin group were decreased(P<0.05),while the apoptosis rate and miR-16-5p expression were increased(P<0.05).Conclusion:Ponicidin combined with radiotherapy has synergistic effect on the apoptosis of lung cancer A549 cells,and the mechanism may be related to promoting the expression of miR-16-5p and then inhibiting the expression of WEE1 by ponicidin.

关 键 词：冬凌草乙素 肺癌 放疗 微小RNA-16-5p/丝氨酸/苏氨酸蛋白激酶1信号轴 

分 类 号：R965.1[医药卫生—药理学]