FGFR1抑制剂PD173074经诱导自噬逆转人肺腺癌HCC827细胞对厄罗替尼耐药
| 作者: |
1刘勇世,
1张志培,
1赖远阳,
1李小飞,
1王小平,
1雷杰
1 空军军医大学唐都医院胸外科，西安 710038 |
| 通讯: |
王小平
Email: wangxiaopingmd@163.com 雷杰 Email: leijiemd@163.com |
| DOI: | 10.3978/j.issn.2095-6959.2019.11.001 |
| 基金: | 陕西省自然科学基础研究计划资助项目(2016JM8009)。 |
摘要
目的：阐明人肺腺癌细胞HCC827厄罗替尼耐药后FGFR1的表达及其与自噬的关系，并探讨FGFR1抑制剂PD173074对肿瘤细胞厄罗替尼耐药的影响。方法：通过低剂量反复刺激法构建HCC827厄罗替尼耐药细胞株(HCC827/ER)；应用CCK-8法检测细胞增殖能力；PE Annexin V/7-AAD双染色法测定细胞凋亡率；RT-qPCR检测FGFR1表达水平；蛋白质印迹法检测FGFR1蛋白及自噬标志物含量。结果：HCC827细胞厄罗替尼耐药后增殖增强(P<0.001)、凋亡减弱(P<0.0001)；耐药细胞FGFR1及编码蛋白表达升高(P<0.0001)。FGFR1抑制剂PD173074可致HCC827/ER增殖减弱(P<0.001)、凋亡增多(P=0.0002)。HCC827/ER细胞LC3-II较HCC827降低(P<0.0001)、p62升高(P<0.001)；PD173074处理后HCC827/ER中LC3-II较处理前升高(P<0.0001)、p62较前降低(P<0.0001)。PD173074与自噬抑制剂HCQ共处理HCC827ER后细胞增殖增强(P=0.013)、凋亡减弱(P=0.0257)。结论： HCC827细胞厄罗替尼耐药后，FGFR1表达升高、细胞自噬下调；而FGFR1抑制剂PD173074可重新诱导肿瘤细胞自噬，从而逆转HCC827细胞对厄罗替尼耐药。FGFR1作为旁路激活是厄罗替尼获得性耐药的机制之一；同时应用厄罗替尼及FGFR1抑制剂可改善EGFR-TKIs耐药。
关键词：
非小细胞肺癌；EGFR-TKIs；耐药；FGFR1；自噬
FGFR1 inhibitor PD173074 reverses HCC827 cells resistance to erlotinib via activating autophagy
CorrespondingAuthor: WANG Xiaoping Email: wangxiaopingmd@163.com
DOI: 10.3978/j.issn.2095-6959.2019.11.001
Foundation: The work was supported by the Natural Science Basic Research Plan in Shaanxi Province of China (2016JM8009).
Abstract
Objective: To elucidate the expression of fibroblast growth factor receptor 1 (FGFR1) in human lung adenocarcinoma cell line HCC827 after erlotinib resistance and its relationship with autophagy and to explore the effect of the FGFR1 inhibitor PD173074 on erlotinib-resistant HCC827 cells. Methods: Erlotinib-resistant HCC827 cell lines (HCC827/ER) was established after low-dose chronic exposure of EGFR-mutant HCC827 cells to erlotinib. Cell viability was detected by CCK-8 method. Cell apoptosis was measured by flow cytometry. The mRNA level of FGFR1 was evaluated by RT-qPCR, and FGFR1 protein and autophagic biomarkers were detected by Western blotting. Results: Erlotinib-resistant HCC827 cells showed increased proliferation (P<0.001), decreased apoptosis (P<0.0001), and up-regulated FGFR1 (P<0.0001). PD173074 inhibited the proliferation of HCC827/ER (P<0.001) and promoted apoptosis (P=0.0002). LC3-II in HCC 827/ER cells expressed lower (P<0.0001) and p62 higher (P<0.001), while after PD173074 treatment LC3-II increased (P<0.0001) and p62 decreased (P<0.0001). When co-treated PD173074 with autophagy inhibitor HCQ, HCC827ER cells proliferated more (P=0.013) and the apoptotic rate decreased (P=0.0257). Conclusion: Erlotinib-resistant HCC827 cells up-regulate FGFR1 to inhibit autophagy and then escape from erlotinib injury. FGFR1 inhibitor PD173074 can re-activate the autophagy of cancer cells, thus reversing the resistance of HCC827 cells to erlotinib.
Keywords：
non-small cell lung cancer cell; EGFR-TKIs; drug resistance; FGFR1; autophagy