曲古抑菌素A联合厄洛替尼对肺腺癌细胞株PC-9/ER增殖及凋亡的影响  

Effects of trichostatin A combined with erlotinib on proliferation and apoptosis of lung adenocarcinoma cell line PC-9/ER

作　　者：张群成 刘庆亮[1] 安云霞[1] ZHANG Quncheng;LIU Qingliang;AN Yunxia(Department of Respiratory and Critical Care Medicine,Henan Provincial People′s Hospital,Zhengzhou 450000,Henan Province,China)

机构地区：[1]河南省人民医院呼吸与危重症医学科,河南郑州450000

出　　处：《新乡医学院学报》2020年第5期404-409,共6页Journal of Xinxiang Medical University

基　　金：河南省医学科技攻关项目(编号:201802040)。

摘　　要：目的探讨曲古抑菌素A联合厄洛替尼对获得性耐厄洛替尼肺腺癌细胞株PC-9/ER增殖、凋亡及耐药性的影响。方法取对数生长期PC-9/ER细胞,接种于96孔培养板,将细胞分为0.0、2.5、5.0、10.0、20.0μmol·L^-1厄洛替尼组和曲古抑菌素A+0.0μmol·L^-1厄洛替尼组、曲古抑菌素A+2.5μmol·L^-1厄洛替尼组、曲古抑菌素A+5.0μmol·L^-1厄洛替尼组、曲古抑菌素A+10.0μmol·L^-1厄洛替尼组、曲古抑菌素A+20.0μmol·L^-1厄洛替尼组;0.0、2.5、5.0、10.0、20.0μmol·L^-1厄洛替尼组细胞分别加入对应浓度的厄洛替尼100μL,曲古抑菌素A联合厄洛替尼组分别加入250 nmol·L^-1的曲古抑菌素A 100μL和对应浓度的厄洛替尼100μL,48 h后采用四甲基偶氮唑盐法检测各组细胞增殖抑制率。取对数生长期PC-9/ER细胞接种于6孔板,将细胞分为厄洛替尼组和曲古抑菌素A+厄洛替尼组;厄洛替尼组细胞给予20μmol·L^-1厄洛替尼100μL,曲古抑菌素A+厄洛替尼组细胞给予250 nmol·L^-1曲古抑菌素A 100μL和20μmol·L^-1厄洛替尼100μL进行处理,24 h后采用流式细胞术检测2组细胞周期分布情况。取对数期PC-9/ER细胞分为空白对照组、曲古抑菌素A组、厄洛替尼组、曲古抑菌素A+厄洛替尼组;空白对照组细胞给予二甲基亚砜100μL,曲古抑菌素A组细胞给予250 nmol·L^-1曲古抑菌素A 100μL,厄洛替尼组细胞给予20μmol·L^-1厄洛替尼100μL,曲古抑菌素A+厄洛替尼组细胞给予250 nmol·L^-1曲古抑菌素A 100μL和20μmol·L^-1厄洛替尼100μL处理细胞,24 h后采用免疫荧光细胞化学法检测4组细胞中E-钙黏蛋白(E-cadherin)的表达并观察PC-9/ER细胞凋亡情况,Western blot法检测4组细胞中磷酸化表皮生长因子受体(p-EGFR)、E-cadherin及Cleaved-caspase-3蛋白的表达。结果不同浓度厄洛替尼联合曲古抑菌素A对PC-9/ER细胞增殖的抑制作用均强于单用厄洛替尼组(P<0.05)。不同浓度厄洛替尼联合曲古抑菌素A�Objective To observe the effect of trichostatin A combined with erlotinib on proliferation,apoptosis and drug resistance of acquired erlotinib resistant lung adenocarcinoma cell line of PC-9/ER.Methods The PC-9/ER cells in logarithmic growth period were inoculated into 96-well culture plate,then the cells were divided into 0.0,2.5,5.0,10.0,20.0μmol·L^-1 erlotinib group and trichostatin A+0.0μmol·L^-1 erlotinib group,trichostatin A+2.5μmol·L^-1 erlotinib group,trichostatin A+5.0μmol·L^-1 erlotinib group,trichostatin A+10.0μmol·L^-1 erlotinib group,trichostatin A+20.0μmol·L^-1 erlotinib group.The cells in 0.0,2.5,5.0,10.0,20.0μmol·L^-1 erlotinib group were added with the corresponding concentration of erlotinib 100μL respectively,and the cells in the combination group of trichostatin A and erlotinib were added with 250 nmol·L^-1 trichostatin A 100μL and the corresponding concentration of erlotinib 100μL respectively.After 48 hours,the inhibition rate of cell proliferation in above each group was determined by methyl thiazolyl tetrazolium.The PC-9/ER cells in logarithmic growth period were inoculated into 6-well culture plate,then the cells were divided into erlotinib group and trichostatin A+erlotinib group.The cells in erlotinib group were added with 20.0μmol·L^-1 erlotinib 100μL;the cells in erlotinib+trichostatin A group were added with 250 nmol·L^-1 trichostatin A 100μL and 20μmol·L^-1 erlotinib 100μL;after 24 hours,the cell cycle distribution in the two groups was detected by flow cytometry.The PC-9/ER cells in logarithmic growth period were divided into blank control group,trichostatin A group,erlotinib group and trichostatin A+erlotinib group.The cells in control group were added with 100μL dimethyl sulfoxide,the cells in trichos tatin A group were added with 250 nmol·L^-1 trichostatin A 100μL,the cells in erlotinib group were added with 20μmol·L^-1 erlotinib 100μL,the cells in trichostatin A+erlotinib group were added with 250 nmol·L^-1 trichostatin A 100μL and 20μmol·L^-

关 键 词：曲古抑菌素A 厄洛替尼 E-钙黏蛋白 磷酸化表皮生长因子受体 

分 类 号：R734.2[医药卫生—肿瘤]