不同浓度华蟾素与吉非替尼联合应用对人非小细胞肺腺癌细胞株A549增殖、凋亡的影响及其机制  

Effect of different concentrations of cinobufacini combined with gefitinib on proliferation and apoptosis of human non-small-cell lung adenocarcinoma cell line A549 and its mechanism

作　　者：马荣辉 董春岚 韩有溪 曹国磊[1] 罗琴[1] MA Ronghui;DONG Chunlan;HAN Youxi;CAO Guolei;LUO Qin(The Third Affiliated Tumor Hospital of Xinjiang Medical University,Urumchi 830000,China)

机构地区：[1]新疆医科大学第三附属肿瘤医院

出　　处：《山东医药》2019年第36期27-31,共5页Shandong Medical Journal

基　　金：新疆维吾尔自治区自然科学基金资助项目(2017D01C411)

摘　　要：目的观察不同浓度华蟾素联合吉非替尼对人非小细胞肺腺癌细胞株A549增殖、凋亡的影响,并探讨其作用机制。方法将A549细胞分为吉非替尼组、华蟾素组、联合药物组和对照组,吉非替尼组分别加入1、5、10、20、40μmol/L的吉非替尼,记为A1、A2、A3、A4、A5组;华蟾素组分别加入0.005、0.01、0.05、0.1、0.5 mg/mL的华蟾素,记为B1、B2、B3、B4、B5组;联合药物组分别加入1μmol/L吉非替尼+0.005 mg/mL华蟾素、5μmol/L吉非替尼+0.01 mg/mL华蟾素、10μmol/L吉非替尼+0.05 mg/mL华蟾素、20μmol/L吉非替尼+0.01 mg/mL华蟾素、40μmol/L吉非替尼+0.5 mg/mL华蟾素,记为C1、C2、C3、C4、C5组;对照组加入等量DMSO。培养24、48、72 h时,采用MTT法观察各组细胞增殖能力(以OD值表示细胞增殖能力),采用流式细胞术测算A5、B5、C5和对照组细胞凋亡率,采用Western Blotting法检测A5、B5、C5和对照组细胞c-met蛋白。结果A1、A2、A3、A4、A5组细胞培养24 h时的OD值分别为0.410±0.030、0.371±0.037、0.322±0.010、0.284±0.025、0.196±0.020,B1、B2、B3、B4、B5组分别为0.463±0.021、0.454±0.017、0.472±0.013、0.460±0.009、0.452±0.028,C1、C2、C3、C4、C5组分别为0.412±0.016、0.359±0.023、0.306±0.035、0.249±0.020、0.174±0.024,对照组为0.458±0.025;其中,吉非替尼组、联合药物组与对照组相比,P均<0.05;吉非替尼组、联合药物组内各浓度组间相比,P均<0.05;C2、C3、C4、C5组与相同浓度的A2、A3、A4、A5组和B2、B3、B4、B5组相比,P均<0.05。A1、A2、A3、A4、A5组细胞培养48 h时的OD值分别为0.541±0.010、0.453±0.016、0.396±0.011、0.344±0.014、0.261±0.011,B1、B2、B3、B4、B5组分别为0.688±0.035、0.687±0.018、0.677±0.018、0.641±0.036、0.556±0.019,C1、C2、C3、C4、C5组分别为0.526±0.033、0.431±0.019、0.376±0.027、0.296±0.022、0.209±0.027,对照组为0.684±0.019;其中,吉非替尼组、B3组、B4组、B5组、联合药物组与对照组Objective To observe the effects of different concentrations of cinobufacini combined with gefitinib on the proliferation and apoptosis of non-small-cell lung adenocarcinoma A549 cells and to discuss its mechanism.Methods A549 cells were divided into the gefitinib group,the cinobufacini group,the drug combination group and the control group.In the gefitinib group,1,5,10,20 and 40 μmol/L gefitinib were added,and marked as the group A1,group A2,group A3,group A4 and group A5;in the cinobufacini group,0.005,0.01,0.05,0.1 and 0.5 mg/mL cinobufacini were add ed separately,and marked as group B1,group B2,group B3,group B4 and group B5;in the drug combination group,1 μmol/L gefitinib+0.005mg/mL cinobufacini,5 μmol/L gefitinib+0.01 mg/mL cinobufacini,10 μmol/L gefitinib+0.05 mg/mL cinobufacini,20 μmol/L gefitinib+0.01 mg/mL cinobufacini,40 μmol/L gefitinib+0.5mg/mL cinobufaci-ni were added separately,and marked as the group C1,group C2,group C3,group C4 and group C5;in the control group,equivalent DMSO was added.At 24,48,and 72 h after culture,the cell proliferative ability(expressed by the OD value)was observed by MTT,the apoptosis rates of group A5,group B5,group C5 and the control group were calculated by flow cytometry,and the cell c-met protein of group A5,group B5,group C5 and the control group was detected by Western Blotting.Results At 24 h,the OD values of group A1,group A2,group A3,group A4,and group A5 were 0.410±0.030,0.371±0.037,0.322±0.010,0.284±0.025,and 0.196±0.020;the OD values of group B1,group B2,group B3,group B4,and group B5 were 0.463±0.021,0.454±0.017,0.472±0.013,0.460±0.009,0.452±0.028;the OD values of group C1,group C2,group C3,group C4,and group C5 were 0.412±0.016,0.359±0.023,0.306±0.0350.249±0.0200.174±0.024;the OD value of the control group was 0.458±0.025;significant differ ence was found between the control group and the gefitinib group and the drug combination group between the gefitinib group and the drug combination group and between the group C2 group C3 group C4 a

关 键 词：华蟾素 吉非替尼 肺癌 非小细胞肺癌 非小细胞肺腺癌 C-MET蛋白 细胞实验 

分 类 号：R73[医药卫生—肿瘤]