摘要
目的探究芍药苷介导SAT1对非小细胞肺癌细胞对培美曲塞敏感性的影响并分析其机制。方法不同浓度芍药苷处理A549细胞,CCK-8检测细胞活力并确定后续实验药物浓度。将细胞分为对照组(Control组)、芍药苷组(Paeoniflorin组)、芍药苷+敲减对照组(Paeoniflorin+sh-NC组)和芍药苷+敲减SAT1组(Paeoniflorin+sh-SAT1组),Western blot验证转染效率;采用CCK-8检测不同浓度培美曲塞处理对各组A549细胞的细胞活力的影响;ROS探针染色检测各组A549细胞中ROS水平;亚铁离子含量检测试剂盒检测各组A549细胞中铁含量;ELISA检测各组A549细胞中MDA和4-HNE含量以及SOD活性。结果芍药苷剂量依赖降低A549细胞的细胞活力,且20μmol/L芍药苷处理的A549细胞的细胞活力最低。芍药苷处理上调A549细胞中SAT1表达,且敲减SAT1逆转芍药苷对培美曲塞处理后A549细胞的细胞活力的下调作用。芍药苷处理上调非小细胞肺癌细胞中ALOX15表达,增加细胞中ROS水平、铁含量以及MDA和4-HNE含量,并降低细胞中SOD活性。敲减SAT1后,芍药苷处理的非小细胞肺癌中ALOX15表达下调,ROS水平、铁含量以及MDA和4-HNE含量降低,SOD活性升高。结论芍药苷通过上调SAT1表达增加非小细胞肺癌细胞对培美曲塞的敏感性。
Objective To explore the effect of paeoniflorin-mediated SAT1 on the sensitivity of non small cell lung cancer cells to pemetrexed and analyze its mechanism.Methods A549 cells were treated with different concentrations of paeoniflorin,and CCK-8 was used to detect the cell viability and determine the drug concentration in the follow up experiment.Cells were divided into control group(Control group),paeoniflorin group(Paeoniflorin group),paeoniflorin+knock-down control group(Paeoniflorin+sh-NC group)and paeoniflorin+knock-down SAT1 group(Paeoniflorin+sh-SAT1 group).Western blot was used to verify transfection efficiency.CCK-8 was used to detect the effects of different concentrations of pemetrexed on the cell viability of A549 cells in each group.ROS probe staining was used to detect the level of ROS in A549 cells in each group.Ferrous ion content detection kit was used to detect the iron content in A549 cells in each group.The contents of MDA and 4-HNE and SOD activity in A549 cells of each group were detected by ELISA.Results Paeoniflorin decreased the cell viability of A549 cells in a dose-dependent manner,and the cell viability of A549 cells treated with 20μmol/L paeoniflorin was the lowest.Paeoniflorin treatment up-regulated the expression of SAT1 in A549 cells,and knocked down SAT1 reversed the down-regulation of paeoniflorin on the cell viability of A549 cells treated with pemetrexed.Paeoniflorin treatment up regulated the expression of ALOx15 in non-small cell lung cancer cells,increased the levels of ROS,iron,MDA and 4-HNE in cells,and decreased the SOD activity in cells.After knocking down SAT1,the expression of ALOX15 in non small cell lung cancer treated with paeoniflorin was down regulated,the levels of ROS,iron,MDA and 4-HNE were decreased,and the activity of SOD was increased.Conclusion Paeoniflorin increases the sensitivity of non small cell lung cancer cells to pemetrexed by up regulating SAT1 expression.
作者
赵睿暄
杨微
陈妍
侯丽丽
ZHAO Rui-xuan;YANG Wei;CHEN Yan;HOU Li-li(Department of Respiratory and Critical Care Medicine,General Hospital of Northern Theater Command of PLA,Shenyang 110016,China)
出处
《解剖科学进展》
2025年第5期639-642,646,共5页
Progress of Anatomical Sciences
基金
辽宁省科学技术计划项目(2021JH2/10300102)。
