摘要
目的探讨苦参碱通过Hedgehog信号通路对肺癌A549细胞顺铂(cisplatin,DDP)化疗敏感性的影响。方法用噻唑蓝(thiazolyl blue,MTT)检测苦参碱的浓度对A549细胞增殖的影响,计算20%致死浓度(20%lethal concentration,LC20);将对数期生长A549细胞分为对照组、苦参碱组(LC20苦参碱干预)、Hedgehog信号通路抑制剂(Hedgehog pathway inhibitor,HPI-4)组(5μmol·L^(−1) HPI-4干预)、DDP组(5 mg·L^(−1) DDP干预)、DDP+苦参碱组(5 mg·L^(−1) DDP和LC20苦参碱共同干预)、DDP+苦参碱+HPI-4组(5 mg·L^(−1) DDP、LC20苦参碱、5μmol·L^(−1) HPI-4共同干预),每组设置3个复孔。用MTT法和流式细胞术检测干预48 h后各组细胞的存活率和凋亡率,用蛋白印迹法(Western blotting)检测各组细胞Hedgehog信号通路相关蛋白音猬因子(sonic hedgehog,SHH)、补缀同源物1(patched1,Ptch1)、神经胶质瘤相关癌基因同源蛋白1(glioma-associated oncogene homolog 1,Gli1)的表达。结果与对照组比较,不同质量浓度的苦参碱干预组细胞存活率均降低(P<0.05),且细胞存活率随苦参碱质量浓度升高而降低(P<0.05)。与对照组比较,苦参碱组、HPI-4组、DDP组、DDP+苦参碱组、DDP+苦参碱+HPI-4组的细胞存活率均降低,且DDP+苦参碱+HPI-4组<DDP+苦参碱组<DDP组<苦参碱组、HPI-4组(P<0.05);细胞凋亡率结果与细胞存活率结果相反。与对照组比较,苦参碱组、HPI-4组、DDP组、DDP+苦参碱、DDP+苦参碱+HPI-4组Hedgehog信号通路SHH、Ptch1、Glis1蛋白的相对表达量均降低,且DDP+苦参碱+HPI-4组<HPI-4组<DDP+苦参碱组<DDP组、苦参碱组(P<0.05)。结论苦参碱可增加肺癌A549细胞DDP化疗敏感性,其作用机制可能与抑制Hedgehog信号通路有关。
Objective To investigate the effect of matrine on the chemotherapy sensitivity to cisplatin(DDP)in lung cancer A549 cells through Hedgehog signaling pathway.Methods Thiazolyl blue(MTT)method was used to detect the effect of matrine at different concentrations on the proliferation of A549 cells,and the 20%lethal concentration(LC20)was calculated.A549 cells grown in log phase were divided into control group,matrine group(treated with LC20 matrine),Hedgehog signaling pathway inhibitor group(HPI-4)group(treated with 5μmol·L^(-1) HPI-4),DDP group(treated with 5 mg·L^(-1) DDP),DDP+matrine group(treated with 5 mg·L^(-1) DDP and LC20 matrine)and DDP+matrine+HPI-4 group(treated with 5 mg·L^(-1) DDP,LC20 matrine and 5μmol·L^(-1) HPI-4),each group had 3 replicate holes.MTT method and flow cytometry were used to detect the survival rate and apoptosis rate of cells in each group after 48 hours of intervention.Western blotting was used to detect the expression of Hedgehog signaling pathway related proteins sonic hedgehog(SHH),patched1(Ptch1)and glioma-associated oncogene homolog 1(Glis1)in each group.Results Compared with the control group,the cell survival rate of the intervention group with different concentrations of matrine was reduced(P<0.05),and the cell survival rate decreased with the increase of matrine dose(P<0.05).Compared with the control group,the cell survival rate of the matrine group,HPI-4 group,DDP group,DDP+matrine group and DDP+matrine+HPI-4 group decreased,and the DDP+matrine+HPI-4 group<DDP+Matrine group<DDP group<matrine group and HPI-4 group(P<0.05).The result of apoptosis rate was opposite to the above.Compared with the control group,the relative expressions of Hedgehog signaling pathway SHH,Ptch1,Glis1 proteins in the matrine group,HPI-4 group,DDP group,DDP+matrine group,and DDP+matrine+HPI-4 group were all decreased,and DDP+matrine+HPI-4 group<HPI-4 group<DDP+matrine group<DDP group and matrine group(P<0.05).Conclusion Matrine can increase the sensitivity of lung cancer A549 cells to DDP chemotherapy,and its mechanism of action may be related to the inhibition of Hedgehog signaling pathway.
作者
赵迎杰
夏松长
聂书慧
王继芳
ZHAO Yingjie;XIA Songchang;NIE Shuhui;WANG Jifang(Pharmacy Department of Traditional Chinese Medicine,Xiangcheng Hospital of Traditional Chinese Medicine,Xiangcheng 466200,China;Department of Respiratory,He’nan Chest Hospital,Zhengzhou 450000,China)
出处
《西北药学杂志》
CAS
2024年第5期61-67,共7页
Northwest Pharmaceutical Journal
基金
2019年河南省医学科技攻关计划项目(编号:LHGJ20190727)。
