脂多糖对核转录因子-κB的激活在人肺非小细胞肺癌获得性耐药细胞中的作用  

Lipopolysaccharide confers to the acquired gefitinib resistance in human non-small cell lung cancer by activating nuclear-κB

作　　者：臧家兰[1] 胡勇[2] 秦晓冰[3] 严大理 吴建中[2] 马蓉[2] 冯继锋[2] 

机构地区：[1]哈尔滨市第一医院肿瘤科,150010 [2]南京医科大学附属肿瘤医院,210009 [3]徐州市第一人民医院肿瘤内科,221000

出　　处：《国际免疫学杂志》2017年第3期238-242,共5页International Journal of Immunology

基　　金：国家自然科学基金资助项目（81372396）

摘　　要：目的 本研究旨在通过检测脂多糖（lipopolysaccharide ,LPS）作用后人非小细胞肺癌耐药株细胞内核转录因子（nuclear factor-κB,NF）-κB p65表达量的变化,探讨LPS作用下NF-κB表达量的增高与吉非替尼获得性耐药产生的相关性.方法 成功诱导非小细胞肺癌细胞株HCC827对吉非替尼产生稳定的获得性耐药后,按设置的分组用LPS及吉非替尼处理细胞,采用CCK-8法检测处理后吉非替尼对细胞的半数抑制浓度（IC50值）,并与各自对照组相比较.细胞增殖曲线显示LPS作用对细胞增殖的影响.采用克隆形成实验显示LPS作用后对细胞增殖的影响.采用蛋白质印迹法检测LPS及吉非替尼处理后耐药株HCC827/GR-8-1细胞中凋亡相关蛋白caspase-3、Bcl-2等蛋白的表达量,并检测NF-κB蛋白的表达量,探讨LPS作用后激活的NF-κB对细胞吉非替尼敏感性的调节.结果 LPS处理后,可以显著降低耐药细胞对吉非替尼的敏感性,细胞克隆形成实验及增殖曲线显示LPS并无明显促进细胞的增殖作用,但可以降低细胞对吉非替尼的敏感性.蛋白质免疫印迹结果显示,与对照组相比较,LPS可抑制Caspase-3等相关促凋亡蛋白的表达,而相应凋亡抑制蛋白Bcl-2的表达上升.LPS处理48 h后NF-κB的表达量显著上调.LPS通过调节NF-κB表达量上升而参与细胞耐药机制的产生.结论 经LPS作用后的非小细胞肺癌细胞对吉非替尼敏感性降低,LPS处理后细胞的NF-κB表达量显著增高,增高的NF-κB表达量与细胞对吉非替尼的药物敏感性下降相关.The study was to confirm that lipopolysaccharide （LPS） could attenuate the gefitinib sensitivity of gefitinib resistance HCC827/GR-8-1 cell line and further to confirm that LPS regulate the gefitinib sensitivity by increasing the expression level of nuclear factor（NF）-κB.We successfully established the gefitinib resistance HCC827/GR-8-1 cell line.Then,cells were treated with LPS and/or gefitinib.To investigate the effect of LPS on non-small cell lung caner cell proliferation,we performed cell counting Kit-8 （CCK8） assay.Then,we conducted the colony formation assay and proliferation curve assay.The levels of caspase-3,bcl-2,and NF-κB were measured by Western blot analysis.Results show that we successfully established the EGFR-TKI-resistant lung cancer cells.Colony formation assay and proliferation curve assay indicated that LPS had little effect on the proliferation and colony formation of HCC827/GR-8-1 cells.But LPS could significantly up-regulate the resistance HCC827/GR-8-1 cells to gefitinib LPS markedly increased the expression level of NF-κB in HCC827/GR-8-1 cells.Additionally,the up-regulation of NF-κB correlated with cells＇ attenuated sensitivity to gefitinb.

关 键 词：非小细胞肺癌 脂多糖 吉非替尼耐药 核转录因子-ΚB 

分 类 号：R734.2[医药卫生—肿瘤]