NEDD4促进非小细胞肺癌继发厄洛替尼耐药

发布时间：2018-04-06 08:00

本文选题：NEDD　切入点：NSCLC　出处：《第三军医大学学报》2017年15期

【摘要】：目的探讨NEDD4(neural precursor cell expressed developmentally downregulated protein 4)在非小细胞肺癌(non-small cell lung cancer,NSCLC)继发厄洛替尼耐药中的作用。方法以HCC827细胞及耐厄洛替尼的HCC827(HCC827/ER)细胞为工具,CCK-8法检测HCC827/ER细胞的耐药指数;实时荧光定量PCR(quantitative real-time,q PCR)及Western blot检测厄洛替尼处理48 h后,NEDD4 mRNA和蛋白在两种细胞中的表达及PI3K/AKT信号通路活化情况。NEDD4小干扰RNA(si NEDD4)转染HCC827/ER细胞,比较处理前后HCC827/ER细胞的厄洛替尼IC50变化以及PI3K/AKT信号通路活化情况。裸鼠成瘤实验在活体水平上进一步验证NEDD4在NSCLC继发厄洛替尼耐药中的作用。结果HCC827/ER细胞的耐药指数为(118.23±23.77);HCC827/ER细胞NEDD4 mRNA和蛋白以及PI3K/AKT信号通路活化水平均高于HCC827细胞;HCC827/ER细胞成功转染si NEDD4后,转染组的PI3K/AKT信号通路活化水平降低,且厄洛替尼IC50值明显低于对照组(P0.05)。裸鼠成瘤实验中转染组肿瘤对厄洛替尼的敏感性明显增加,与阴性对照组比较,药物处理组肿瘤生长受到明显的抑制。结论NEDD4通过激活PI3K/AKT信号通路促进NSCLC继发厄洛替尼耐药。
[Abstract]:Objective to investigate the role of NEDD4(neural precursor cell expressed developmentally downregulated protein 4 in the development of erlotinib resistance secondary to non-small cell lung cancer in non-small cell lung cancer.Methods the drug resistance index of HCC827/ER cells was measured by CCK-8 method using HCC827 cells and HCC827- HCC827 / ER-ERcells of erlotinib as a tool.Real-time PCR(quantitative real-time PCR and Western blot were used to detect the expression of Neddd 4 mRNA and protein in two kinds of cells and the activation of PI3K/AKT signaling pathway. NEDD4 small interfering RNA(si NEDD4 was transfected into HCC827/ER cells after 48 h of erlotinib treatment.The changes of erlotinib IC50 and the activation of PI3K/AKT signaling pathway in HCC827/ER cells were compared before and after treatment.Tumorigenesis in nude mice further demonstrated the role of NEDD4 in erlotinib resistance secondary to NSCLC in vivo.Results the drug resistance index of HCC827/ER cells was 118.23 卤23.77% HCC827 / ER cells. The activation levels of NEDD4 mRNA, protein and PI3K/AKT signaling pathway in ER cells were higher than those in HCC827 cells after successful transfection of HCC827 cell line HCC827 / ER cells. The activation level of PI3K/AKT signaling pathway in transfection group was lower than that in HCC827 cell line.The IC50 value of erlotinib was significantly lower than that of control group (P 0.05).The sensitivity of the transfected group to erlotinib was significantly increased in nude mice. Compared with the negative control group, the tumor growth of the drug treated group was significantly inhibited.Conclusion NEDD4 promotes erlotinib resistance secondary to NSCLC by activating PI3K/AKT signaling pathway.
【作者单位】：第三军医大学新桥医院全军肿瘤研究所;
【基金】：国家自然科学基金面上项目(81672841) 重庆市科委重点攻关课题(2011AB5032)~~
【分类号】：R979.1

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