阿美替尼对非小细胞肺癌细胞增殖、侵袭、上皮间质转化和干性的作用  

Effects of almonertinib on proliferation,invasion,epithelial mesenchymalt ransformation and stemness of non-small cell lung cancer cells

作　　者：谢友琴 高振 张亮[1] XIE You-qin;GAO Zhen;ZHANG Liang(Department of Oncology,Nantong First People sHospital,Nantong 226000,Jiangsu Province,China;Department of Radiotherapy,Nantong First People sHospital,Nantong 226000,Jiangsu Province,China)

机构地区：[1]南通市第一人民医院肿瘤科,江苏南通226000 [2]南通市第一人民医院放疗科,江苏南通226000

出　　处：《中国临床药理学杂志》2025年第5期660-664,共5页The Chinese Journal of Clinical Pharmacology

基　　金：南通市卫生健康委员会科研基金资助项目(MS2023030)。

摘　　要：目的研究阿美替尼通过调控微小核糖核酸-22(miR-22)靶向泛素结合酶E2O(UBE2O)对非小细胞肺癌(NSCLC)细胞的增殖、侵袭、上皮间质转化和干性的影响。方法将人肺腺癌细胞系NCI-H1975随机分为对照组(无任何处理正常培养)、中剂量组(4μmol·L^(-1)阿美替尼处理)、抑制药阴性对照(转染NC inhibitor)组(4μmol·L^(-1)阿美替尼+miR-22抑制药阴性对照处理)、miR-22抑制药(miR-22 inhibitor)组(4μmol·L^(-1)阿美替尼+miR-22抑制药处理)、miR-22 inhibitor+短发卡阴性对照(sh-NC)组(4μmol·L^(-1)阿美替尼+miR-22抑制药+短发卡阴性对照处理)、miR-22 inhibitor+短发卡UBE2O(sh-UBE2O)组(4μmol·L^(-1)阿美替尼+miR-22抑制药+短发卡UBE2O处理)。用实时荧光定量聚合酶链反应(qRT-PCR)检测miR-22表达,用蛋白质印迹法检测八聚体结合转录因子4(OCT4)、性别决定区域Y盒基因2(SOX2)和Nanog同源框的表达水平,用5-乙炔基-2′-脱氧尿嘧啶核苷(EdU)实验检测细胞增殖率,用Transwell实验检测细胞侵袭能力。结果对照组和中剂量组的miR-22相对表达水平分别为1.00±0.16和2.07±0.34;对照组、中剂量组、NC inhibitor组和miR-22 inhibitor组的OCT4蛋白相对表达水平分别为1.00±0.16、0.63±0.09、0.67±0.08和0.94±0.12,SOX2蛋白相对表达水平分别为1.00±0.14、0.48±0.06、0.54±0.07和0.81±0.11,Nanog蛋白相对表达水平分别为1.00±0.18、0.45±0.08、0.52±0.11和0.89±0.15。对照组、中剂量组、NC inhibitor组、miR-22 inhibitor组、miR-22 inhibitor+sh-NC组和miR-22 inhibitor+sh-UBE2O组的细胞增殖率分别为(87.29±10.73)%、(41.65±7.82)%、(48.47±6.54)%、(79.31±10.26)%、(84.50±11.98)%和(36.82±6.79)%,细胞侵袭数目分别为(156.92±24.81)、(72.53±9.06)、(64.41±10.75)、(143.27±26.19)、(137.58±21.46)和(75.23±11.94)个。对照组的上述指标与中剂量组,NC inhibitor组的上述指标与miR-22 inhibitor组,miR-22 inhibitor+sh-NC组的上述指标与miR-22 inhibitor+sh-UBObjective To investigate the effects of almonertinib on the proliferation,invasion,epithelial-mesenchymal transition(EMT),and stemness of non-small-cell lung cancer(NSCLC)cells by regulating microRNA-22(miR-22)targeting ubiquitin-conjugating enzyme E2O(UBE2O).Methods Human lung adenocarcinoma cell line NCI-H1975 was randomly divided into control group(normal culture with no treatment),medium-dose group(4μmol·L^(-1)almonertinib),inhibitor negative control(NC inhibitor)group(4μmol·L^(-1)almonertinib+miR-22 inhibition negative control),miR-22 inhibitor group(4μmol·L^(-1)almonertinib+miR-22 inhibition),miR-22 inhibitor+short hairpin negative control(sh-NC)group(4μmol·L^(-1)almonertinib+miR-22 inhibition+short hairpin negative control),and miR-22 inhibitor+short hairpin UBE2O(sh-UBE2O)group(4μmol·L^(-1)almonertinib+miR-22 inhibition+short hairpin UBE2O).The expression ofmiR-22 was detected by quantitative real-time polymerase chain reaction(qRT-PCR);Western blot analysis was performed to detect the expression of octamer-binding transcription factor 4(OCT4),SRY-Box transcription factor 2(SOX2),and Nanog;the cell proliferation rate was assessed using the 5-ethynyl-2-deoxyuridine(EdU)assay;and Transwell assays were conducted to evaluate cell invasion ability.Results The expression levels ofmiR-22 in the control group,medium-dose group were 1.00±0.16,2.07±0.34.The expression levels of OCT4 in the control group,medium-dose group,NC inhibitor group,and miR-22 inhibitor group were 1.00±0.16,0.63±0.09,0.67±0.08 and 0.94±0.12,respectively;SOX2 expression levels were 1.00±0.14,0.48±0.06,0.54±0.07and 0.81±0.11;and Nanog expression levels were 1.00±0.18,0.45±0.08,0.52±0.11 and 0.89±0.15.The cell proliferation rates in the control group,medium-dose group,NC inhibitor group,miR-22 inhibitor group,miR-22 inhibitor+sh-NC group,and miR-22 inhibitor+sh-UBE2O group were(87.29±10.73)%,(41.65±7.82)%,(48.47±6.54)%,(79.31±10.26)%,(84.50±11.98)%and(36.82±6.79)%,respectively.The numbers of invading cells were(156

关 键 词：阿美替尼 微小核糖核酸-22 泛素结合酶E2O 非小细胞肺癌 增殖 侵袭 上皮间质转化 干性标志物 

分 类 号：R979.1[医药卫生—药品]