吉非替尼联合安罗替尼对表皮生长因子受体敏感及耐药细胞株的作用研究  

Effects of gefitinib combined with anlotinib on epidermal growthfactor receptor sensitive and resistant cells

作　　者：段方方[1] 周寒丽[1] 刘丹娜[2] 苏田丽 赵晓丽 陈露[1] 刘萌萌[3] 孔天东[1] DUAN Fang-fang;ZHOU Han-li;LIU Dan-na;SU Tian-li;ZHAO Xiao-li;CHEN Lu;LIU Meng-meng;KONG Tian-dong(Department of Medical Oncology,the Third People’s Hospital of Zhengzhou,the Cancer Hospital of Henan University,Zhengzhou 450000,Henan Province,China;Drug Clinical Trial Center,the Third People’s Hospital of Zhengzhou,the Cancer Hospital of Henan University,Zhengzhou 450000,Henan Province,China;Department of Molecular Pathology,the Third People’s Hospital of Zhengzhou,the Cancer Hospital of Henan University,Zhengzhou 450000,Henan Province,China)

机构地区：[1]郑州市第三人民医院、河南大学肿瘤医院,肿瘤内科,河南郑州450000 [2]郑州市第三人民医院、河南大学肿瘤医院药学部,河南郑州450000 [3]郑州市第三人民医院、河南大学肿瘤医院分子病理室,河南郑州450000

出　　处：《中国临床药理学杂志》2023年第8期1162-1166,共5页The Chinese Journal of Clinical Pharmacology

基　　金：河南省科技厅重点科技研发计划基金资助项目(202102310456);河南省医学科技联合共建计划基金资助项目(LHGJ20191028)。

摘　　要：目的研究安罗替尼联合吉非替尼对HCC827、HCC827GR肺癌细胞的杀伤作用。方法细胞分为对照组(不加入任何药物),吉非替尼组(1000 nmol·L^(-1)吉非替尼处理),安罗替尼组(2.0μmol·L^(-1)安罗替尼处理),联合组(先加入2.0μmol·L^(-1)安罗替尼,6 h后再加入1000 nmol·L^(-1)吉非替尼)。以细胞计数法-8(CCK-8)法测定细胞增殖活性;用流式细胞术及Transwell法测定对HCC827、HCC827GR细胞凋亡、侵袭的影响,用蛋白质印迹法测定凋亡相关蛋白的表达。结果HCC827细胞中,对照组、非替尼组、安罗替组尼和联合组的细胞增殖率分别为(97.43±1.65)%、(50.78±2.30)%、(52.54±3.66)%和(35.47±2.55)%;吉非替尼、安罗替尼均能明显降低HCC827细胞增殖,且两者联合使用抑制作用更明显。对耐药的HCC827GR细胞,对照组、非替尼组、安罗替组尼和联合组的细胞增殖率分别为(97.16±2.74)%、(94.14±3.56)%、(58.69±2.64)%和(40.43±2.43)%;单药吉非替尼无明显抑制作用,但联合安罗替尼则能明显降低HCC827GR细胞的增殖和侵袭。HCC827GR细胞中,吉非替尼组、安罗替尼组和联合组凋亡率分别为(10.63±2.24)%、(37.52±4.33)%和(46.93±8.56)%,差异有统计学意义;HCC827GR细胞中,对照组、吉非替尼、安罗替尼和联合组细胞侵袭数分别为148.67±3.21、144.00±5.29、99.33±3.51和44.67±4.73,与对照组相比,吉非替尼组细胞侵袭数无明显变化(P>0.05),而安罗替尼组及联合组均能明显减少侵袭细胞数,联合组更明显。凋亡蛋白比较,对HCC827细胞,吉非替尼组、安罗替尼组和联合组均能降低磷酸化表皮生长因子受体(p-EGFR)、磷酸化蛋白激酶(p-Akt)、磷酸化细胞外调节蛋白激酶(p-ERK)蛋白表达水平,尤其是联合组表达水平降低更明显(P<0.05)。对于HCC827GR细胞,安罗替尼组能降低p-Akt、p-ERK蛋白表达,联合组p-EGFR、p-Akt、p-ERK蛋白表达水平明显降低(P<0.05)。结论吉非替尼联合安罗替尼可协同�Objective To investigate the killing effect of anlotinib combined with gefitinib on HCC827 and HCC827GR lung cancer cells.Methods The cells were divided into control group(without any drug),gefitinib group(1000 nmol·L^(-1) gefitinib),anlotinib group(2.0μmol·L^(-1) anlotinib),and Gefi+Anlo group(2.0μmol·L^(-1) anlotinib was added at first,1000 nmol·L^(-1) gefitinib was added 6 h l ate r).Cell proliferation activity was determined by cell counting kit-8(CCK-8)method.Flow cytometry and Transwell methods were used to determine the effect of apoptosis and invasion on HCC827 and HCC827GR cells,and Western blotting was used to determine the expression of apoptosis-related proteins.Results The proliferation rate of HCC827 cells in control group,gefitinib group,anlotinib group and Gefi+Anlo group were(97.43±1.65)%,(50.78±2.30)%,(52.54±3.66)%and(35.47±2.55)%,respectively,both gefitinib and anrotinib could significantly reduce the proliferation of HCC827 cells,and the combined use of them had more obvious inhibitory effects.For drug-resistant HCC827GR cells,the cell proliferation rates of the four groups were(97.16±2.74)%,(94.14±3.56)%,(58.69±2.64)%and(40.43±2.43)%;gefitinib alone had no significant inhibitory effect,but combined with anrotinib could significantly reduce the proliferation and invasion of HCC827GR cells.The apoptosis rate of HCC827GR cells in gefitinib group,anlotinib group and Gefi+Anlo group were(10.63±2.24)%,(37.52±4.33)%,(46.93±8.56)%,and the difference was statistically significant.The invasion numbers of control,gefitinib,anlotinib,and Gefi+Anlo groups of HCC827GR cells were 148.67±3.21,144.00±5.29,99.33±3.51,44.67±4.73,compared with control group,the invasion number of gefitinib group had no significant change(P>0.05),while anlotinib group and Gefi+Anlo group could significantly reduce the invasion number of cells,the Gefi+Anlo group was more obvious.Compared with apoptosis proteins,the expressions of phosphorylated epidermal growth factor receptor(p-EGFR),phosphorylated protein k

关 键 词：吉非替尼 安罗替尼 表皮生长因子受体 肺癌 凋亡 

分 类 号：R979.1[医药卫生—药品]