色瑞替尼对肺癌细胞H3122增殖及磷脂酰肌醇3-激酶/蛋白激酶B信号通路的影响  

Effect of ceritinib on proliferation of lung cancer cells H3122 and phosphatidylinositol 3-kinase/protein kinase B signaling pathway

作　　者：侯继院[1] 单国用[1] 王海霞[1] 王作培[2] HOU Jiyuan;SHAN Guoyong;WANG Haixia;WANG Zuopei(Department of Radiation Oncology,People′s Hospital of Zhengzhou,Zhengzhou 450000,Henan Province,China;Department of Cardiothoracic Oncology Surgery,Huaihe Hospital of Henan University,Kaifeng 475000,Henan Province,China)

机构地区：[1]郑州人民医院肿瘤放疗科,河南郑州450000 [2]河南大学淮河医院心胸肿瘤外科,河南开封475000

出　　处：《新乡医学院学报》2020年第7期601-605,共5页Journal of Xinxiang Medical University

基　　金：河南省高等学校重点科研项目(编号:18A320027)。

摘　　要：目的探讨色瑞替尼对肺癌细胞H3122增殖及磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)信号通路的影响。方法将对数生长期H3122细胞随机分为空白组及低、中、高剂量色瑞替尼组,每组设5个复孔。低、中、高剂量色瑞替尼组分别加入色瑞替尼,使每孔终质量浓度分别为16.67、33.33、66.66 mg·L^-1,空白组加入等量的磷酸盐缓冲液。采用甲基噻唑基四唑法检测干预24、48、72 h后各组细胞增殖抑制率,流式细胞术检测干预72 h后各组细胞凋亡率,实时荧光定量聚合酶链式反应检测干预72 h后各组细胞中PI3K、Akt、Caspase-3 mRNA相对表达量,蛋白质免疫印迹法检测干预72 h后各组细胞中PI3K、p-PI3K、Akt、p-Akt、Caspase-3蛋白表达。结果培养24、48、72 h后,低、中、高剂量色瑞替尼组细胞增殖抑制率均高于空白组(P<0.05),中、高剂量色瑞替尼组细胞增殖抑制率高于低剂量组(P<0.05),高剂量色瑞替尼组细胞增殖抑制率高于中剂量组(P<0.05);低、中、高剂量色瑞替尼组细胞增殖抑制率随时间延长均呈显著上升趋势(P<0.05)。培养72 h时,低、中、高剂量色瑞替尼组细胞凋亡率均高于空白组(P<0.05),中、高剂量色瑞替尼组细胞凋亡率高于低剂量组(P<0.05),高剂量色瑞替尼组细胞凋亡率高于中剂量组(P<0.05)。4组细胞中Caspase-3 mRNA相对表达量随色瑞替尼剂量的增加呈上升趋势,组间两两比较差异均有统计学意义(P<0.05);4组细胞中PI3K、Akt mRNA相对表达量比较差异均无统计学意义(P>0.05)。4组细胞中Caspase-3蛋白相对表达量随色瑞替尼剂量增加呈上升趋势,p-PI3K/PI3K、pAkt/Akt随色瑞替尼剂量的增加呈下降趋势,组间两两比较差异均有统计学意义(P<0.05)。结论色瑞替尼可抑制肺癌细胞增殖,促使其凋亡,且呈一定的剂量依赖性,其作用机制可能与调控PI3K、Akt及其下游Caspase-3基因和蛋白表达有关。Objective To investigate the effect of ceritinib on the proliferation of lung cancer cells H3122 and the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)signaling pathway.Methods The H3122 cells in logarithmic phase were randomly divided into blank group,low-dose ceritinib group,medium-dose ceritinib group and high-dose of ceritinib group,and each group was provided with 5 replicate.The ceritinib was added to the low-,medium-and high-dose ceritinib groups so that the final concentration of each well was 16.67,33.33,66.66 mg·L-1,and the blank group was added with the equal amount of phosphate buffer solution(PBS).The cell proliferation inhibition rate was detected by methyl thiazolyl tetrazolium method at 24,48,and 72 hours after intervention in each group.The apoptosis rate at 72 hours after intervention was detected by flow cytometry.The relative expression of PI3K,Akt,and Caspase-3 mRNA in the cells was detected by real-time fluorescent quantitative polymerase chain reaction at 72 hours after intervention.The expression of PI3K,p-PI3K,Akt,p-Akt,Caspase-3 protein in the cells was detected by Western blot at 72 hours after intervention in each group,and the p-PI3K/PI3K,p-Akt/Akt in each group were compared.Results The cell proliferation inhibition rate in the low-,medium-and high-dose ceritinib groups was higher than that in the blank group at 24,48 and 72 hours after intervention(P<0.05).The cell prolifera tion inhibition rate in the medium-and high-dose ceritinib groups was higher than that in the low-dose ceritinib group(P<0.05).The cell proliferation inhibition rate in the high-dose ceritinib group was higher than that in the medium-dose ceritinib group(P<0.05).The cell proliferation inhibition rate in the low-,medium-and high-dose ceritinib groups increased significantly with the extension of the time(P<0.05).The apoptosis rate in the low-,medium-and high-dose groups was higher than that in the blank group(P<0.05).The apoptosis rate in the medium-and high-dose ceritinib groups was lower than that i

关 键 词：色瑞替尼 肺癌细胞 细胞增殖 磷脂酰肌醇3-激酶/蛋白激酶B信号通路 

分 类 号：R734.2[医药卫生—肿瘤]