KRAS突变对非小细胞肺癌增殖的影响及其机制  

Effect of KRAS mutation on the proliferation of NSCLC cells

作　　者：曾诚[1] 姚贝[1] 吴启鹏[1] 刘冰[1] 

机构地区：[1]广东药学院药科学院,广东广州510006

出　　处：《广东药学院学报》2016年第4期503-506,517,共5页Academic Journal of Guangdong College of Pharmacy

基　　金：广东省自然科学基金(2015A030313580)

摘　　要：目的观察在非小细胞肺癌(NSCLC)细胞中KRAS突变对炎症因子IL-6和NADPH氧化酶4(NOX4)表达的影响,并研究由KRAS突变引起的IL-6分泌和NOX4的表达对NSCLC细胞增殖能力的影响。方法分别通过质粒p Babe puro-KRAS^(G12V)和siRNA转染细胞,制备KRAS^(G12V)过表达的Calu3细胞株和KRAS^(G12V)干扰的H411细胞株,采用Western blotting法对NOX4的表达进行检测;采用ELISA法检测IL-6的分泌水平;采用MTT法检测Calu3细胞的增殖能力。结果与对照组相比,经质粒p Babe puro KRAS^(G12V)转染后,Calu3细胞中NOX4的蛋白表达水平和IL-6的分泌水平均显著升高;经siRNA转染干扰KRAS^(G12V)后,H411细胞中NOX4表达和IL-6的分泌量均显著降低;在KRAS^(G12V)过表达的基础上,在Calu3细胞中加入IL-6的中和抗体Siltuximab可降低NOX4的表达,而干扰NOX4后可减少IL-6的分泌;在过表达KRAS^(G12V)的Calu3细胞中,分别干扰NOX4的表达或加入Siltuximab,均可显著抑制细胞的增殖能力。结论在NSCLC细胞中,KRAS的突变能引起IL-6的分泌和NOX4的表达,并通过IL-6和NOX4的相互激活共同促进癌细胞的恶性增殖。Objective To study the effect of KRAS mutation on the expression of IL-6 and NOX4 and identify the role of NOX4 and IL-6 on the cell proliferation in NSCLC cells.Methods KRASG12 Voverexpressed Calu3 cells and KRASG12V-intervened H411 cells were generated by transfection of p Babe puro-KRASG12 Vand KRASG12VsiRNA, respectively.The expression of NOX4 was analyzed by western blotting.The level of IL-6 was determined by ELISA.The ability of Calu3 cell proliferation was analyzed by MTT assay.Results Compared with the control group, the expression of NOX4 and IL-6 were obviously increased in Calu3 cells after transfected with KRASG12 Vplasmid,which was significantly reduced in H411 cells after transfected with KRASG12 VsiRNA.After treatment with siltuximab or NOX4 siRNA, the expression of NOX4 and IL-6 were reduced in Calu3 cells with overexpression of KRASG12 V.Meantime,treatment with siltuximab or NOX4 siRNA inhibited the Calu3 cell proliferation.Conclusion KRAS mutation enhances the expression of IL-6 and NOX4 in NSCLC cells, which may promote the cell proliferation.

关 键 词：KRAS NOX4 IL-6 NSCLC 细胞增殖 

分 类 号：R734.2[医药卫生—肿瘤]