水飞蓟宾增敏克唑替尼抑制间变性淋巴瘤激酶阳性非小细胞肺癌的作用及机制  

Effect and mechanism of silibinin on the inhibition of ALK positive NSCLC cells by sensitizing crizotinib

作　　者：林采余[1] 卢从华[1] 潘永红[1] 焦琳[1] 陈恒屹[1] 李力[1] 何勇[1] 

机构地区：[1]第三军医大学大坪医院呼吸内科,重庆400042

出　　处：《中华肿瘤杂志》2017年第9期650-656,共7页Chinese Journal of Oncology

摘　　要：目的研究水飞蓟宾联合克唑替尼对间变性淋巴瘤激酶（ALK）基因阳性非小细胞肺癌（NSCLC）细胞的增殖抑制作用和机制。方法采用单加水飞蓟宾、单加克唑替尼或两药联合作用于NSCLC细胞株H2228和H3122，以四甲基偶氮唑蓝（MTT）法和克隆形成实验检测细胞的增殖活性，以划痕实验和侵袭实验检测细胞的迁移和侵袭能力，以免疫荧光实验检测细胞上皮间质转化（EMT）标志蛋白E-Cadherin和Vimentin的表达情况，以Westernblot法检测细胞中ALK、P-ALK、E-Cadherin和Vimentin蛋白的表达变化。将H2228细胞悬液注射入裸鼠皮下，构建裸鼠移植瘤模型，观察移植瘤的生长情况。结果MTr法检测结果显示，当水飞蓟宾浓度为100μmol／L时，H2228和H3122细胞的存活率分别为（88．38±4．10）％和（72．27±3．62）％，对细胞的增殖活性影响较小。当100μmol／L的水飞蓟宾与克唑替尼联合使用时，克唑替尼对H2228细胞的I50从（917．10±7．75）nmoL／L下降到（238．73±7．67）nmol／L，对H3122细胞的I50从（472．50＋15．70）nmol／L下降到（206．10±12．01）nmol／L，联合用药后克唑替尼对H2228和H3122细胞的I50明显降低。与对照组H2228细胞比较，100μmol／L水飞蓟宾组、400nmol／L克唑替尼组、100μmol／L水飞蓟宾联合400nmoL／L克唑替尼组H2228细胞的克隆形成率分别为（83．34±2．72）％、（69．42±3．06）％和（27．32±1．42）％；与对照组H3122细胞比较，100μmoL／L水飞蓟宾组、400nmol／L克唑替尼组、100μmol／L水飞蓟宾联合400nmol／L克唑替尼组H2228细胞的克隆形成率分别为（84．45±5．67）％、（45．02±5．83）％和（17．43±3．83）％。水飞蓟宾联合克唑替尼后，H2228和H3122细胞的克隆形成能力均明显下降（均P〈0．01）。迁移和侵袭实验结果显示．水飞蓟宾与克唑替尼联用能明显抑制H2228细胞的迁移和侵袭（均P〈O．01Objective To explore the synergistic effect of silibinin combined with crizotinib on anaplastic lymphoma kinase positive （ALK＋） non-small cell lung cancer （NSCLC） cells and its mechanism. Methods H2228 and H3122 cells were treated with silihinin, crizotinib alone or in combination. Cell proliferation was measured by 3-（4,5-Dimethylthiazol-2-yl）-2,5-diphenyhetrazolium bromide （MTT） assay and colony formation assay. Migration or invasion ability was tested by wound healing assay or transwell assay, respectively. Expressions of E-Cadherin and vimentin protein were examined by immunofluoreseence staining. The protein expressions of ALK, p-ALK, E-Cadherin and Vimentin were detected by western blotting.The anti-cancer effect of silibinin combined with erizotinib in vivo was determined by subcutaneously injecting 2×10^6 H2228 cells into immunodeficient nude mice. Results The result of MTT assay showed that the cell viability of H2228 or H3122 treated with 100 μmoL/L silibinin was （88.38±4.10）% or （72.27±3.62）%, respectively, marginally decreased compared with that of the control. The 50% inhibitory concentration （ICs0） of H2228 cells treated with crizotiuib alone or combined with 100 μmol/L silibinin was （917.10±7.75） nmol/L or （238.73±7.67） nmol/L, respectively. The ICs0 of H3122 cells treated with crizotinib alone or combined with 100 μmol/L silibinin was （472.50±15.70） nmoL/L or （ 206.10±12.01 ） μmol/L, respectively. The IC50 of H2228 and H3122 cells were significantly decreased by combined treatment of crizotinib and silibinin compared to crizotinib treatment alone （P〈0.01）. When compared with the control group, colony forming ratios of H2228 cells were （83.34±2.72）% in 100 μmoL/L silibinin treatment group, （ 69.42±3.06） % in 400 nmoL/L crizotinib treatment group and （ 27.32 ± 1.42） % in combined treatment group. When compared with the control group, colony forming ratios of H3122 cells were （ 84.45 ±5.67 ） % in 100 μmol/L silibinin

关 键 词：克唑替尼 水飞蓟宾 间变性淋巴瘤激酶 癌 非小细胞肺 

分 类 号：R734.2[医药卫生—肿瘤]