蛋白激酶2抑制剂增强盐酸埃克替尼对表皮生长因子受体-酪氨酸激酶抑制剂耐药细胞的增殖抑制作用及机制  

The CK2 inhibitor quninalizarin enhances the anti-proliferative effect of icotinib on EGFR-TKIs- resistant call lines and its underlying mechanisms

作　　者：周瑜[1] 张盛[1] 李珂[2] 李倩雯[1] 周方正[1] 李振宇[1] 马虹[1] 董晓荣[1] 刘莉[1] 伍钢[1] 孟睿[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院肿瘤中心,武汉430023 [2]华中科技大学同济医学院附属协和医院药剂科,武汉430023

出　　处：《中华肿瘤杂志》2016年第2期100-104,共5页Chinese Journal of Oncology

基　　金：国家自然科学基金青年基金（81101690）;武汉市科技局应用基础研究资助项目（2014060101010034）;湖北省自然科学基金（2014cfb403）

摘　　要：目的探讨蛋白激酶2（CK2）抑制剂琨茜素增强盐酸埃克替尼在表皮生长因子受体-酪氨酸激酶抑制剂（EGFR—TKIs）耐药细胞中的增殖抑制作用及机制。方法采用四甲基偶氮唑蓝（MTT）法检测盐酸埃克替尼和琨茜素对不同人肺腺癌细胞的增殖抑制作用，Westernblot法检测盐酸埃克替尼和琨茜素作用后不同人肺腺癌细胞中EGFR下游信号通路主要蛋白的表达水平及磷酸化水平。结果盐酸埃克替尼在人肺腺癌HCC827、H1650、H1975和A549细胞中的Ic。分别为（8．07±2．00）μmol／L、（66．01±6．64）μmol／L、（265．60±9．47）μmol／L和（87．88±6．80）μmol／L，提示HCC827细胞为盐酸埃克替尼敏感细胞，H1650、H1975和A549细胞为盐酸埃克替尼耐药细胞。在50μmol／L盐酸埃克替尼和50μmol／L琨茜素联合作用下，H1650、H1975和A549细胞的存活率分别为（40．64±3．73）％、（65．74±3．27）％和（44．96±0．48）％，明显低于50μmol／L盐酸埃克替尼作用时的细胞存活率[（55．05±1．22）％、（71．98±1．60）％和（61．74±6．18）％]，差异有统计学意义（均P〈0．01）。在100μmol／L盐酸埃克替尼和100μmoL／L琨茜素联合作用下，H1650、H1975和A549细胞的存活率分别为（23．35±0．81）％、（55．70±1．03）％和（33．42±1．33）％，明显低于100μmol／L盐酸埃克替尼作用时的细胞存活率[（40．57±2．65）％、（62．40±2．05）％和（44．97±8．20）％]，差异有统计学意义（均P〈0．01）。Westernblot检测显示，联合使用盐酸埃克替尼和琨茜素作用H1650细胞后，EGFR下游信号通路中蛋白激酶B（Akt）及细胞外调节蛋白激酶（ERK）的表达水平无明显变化，但Akt和ERK磷酸化水平明显降低。结论CK2抑制剂琨茜素可能通过协同抑制细胞增殖相关信号通路中Akt和ERK的磷酸化，增强盐酸埃克替尼在EGFR—TKIs耐药的人肺腺癌细胞�Objective To explore whether quninalizarin, an specific inhibitor of protein kinase CK2, could sensitize icotinib in EGFR-TKIs （epithelial growth factor receptor-tyrosine kinase inhibitor）- resistant cell lines and uncover the underlying mechanisms. Methods MTT assay was performed to evaluate the inhibitory effect of quninalizarin, icotinib or the combination of both on cell proliferation in several lung adenocarcinoma cell lines. Western blot assay was used to assess if combined inhibition of EGFR and protein kinase CK2 by icotinib and quninalizarin, exerts effect on the expression and phosphorylation of major proteins of EGFR signaling pathways. Results The IC50 of HCC827, H1650, H1975 and A549 cells for icotinib were （ 8.07 ± 2.00 ） μmol/L, （ 66.01 ± 6.64 ） μmol/L, （ 265.60± 9.47 ） μmol/L and （ 87.88 ± 6.8 ） μmol/L, respectively, indicating that HCC827 cells are sensitive to icotinib, and the H1650, H1975 and A549 cells are relatively resistant to icotinib. When treated with both quninalizarin and icotinib in the concentration of 50 μmol/L, the viability of H1650, H1975 and A549 cells was （40.64±3.73）%, （65.74± 3.27） % and （44.96±0.48）%, respectively, significantly lower than that of H1650, H1975 and A549 cells treated with 50 μmoL/L ieotinib alone （55.05＋1.22）%, （71.98±1.60）% and （61.74±6.18）%, respectively （P〈0.01 for all）. When treated with both 100 μmol/L quninalizarin and 100 μmol/L icotinib, the viability of H1650, H1975 and A549 ells were （23.35±0.81）%, （55.70±1.03）%, （33.42＋l.33）%,respeetively, significantly lower than the viability of H1650, H1975 and A549 cells treated with 100 txmol/L icotinib alone （40.57±2.65） %, （62.40±2.05） % and （44.97±8.20） %, respectively, （P〈0.01 for all）. The two- way ANOVA analysis showed that compared with the viability of EGFR-TKIs-resistant cells （ H1650, H1975, A549） treated with 50 μmol/L and 100 μmol/L icotinib alone, the viability of cells treated with ico

关 键 词：肺肿瘤 细胞系 肿瘤 细胞存活 盐酸埃克替尼 琨茜素 

分 类 号：R734.2[医药卫生—肿瘤]