Derlin-1介导非小细胞肺癌EGFR-TKI耐药的分子机制研究

EGFR-TKI resistance significantly limits its use in clinical practice. We found that Derlin-1 was overexpressed in lung cancer cell lines with EGFR-TKI resistance. Gefitinib treatment upregulated Derlin-1 levels in lung cancer cells haboring EGFR mutation. Derlin-1 transfection could alleviate the effect of gefitinib on cell growth inhibition and upregulated Akt activity, indicating the role of Akt pathway in Derlin-1 mediated EGFR-TKI resistance. However, its molecular mechanism still needs to be explored. In this study, we will upregulate and knockdown Derlin-1 expression in lung cancer cells and use gene chip, antibody microarray, immunoprecipitation and GST-pulldown to investigate the molecular machanism of derlin-1 mediated EGFR-TKI resistance in lung cancer cells. We will also use animal model and clinical samples to explore the clinical significance of Derlin-1 in predicting EGFR-TKI sensitivity and patient prognosis.

EGFR-TKI的原发和继发性耐药已成为其在肺癌治疗中应用的瓶颈。我们前期研究发现Derlin-1在EGFR-TKI耐药细胞系中高表达，吉非替尼处理能够上调EGFR突变细胞内Derlin-1的表达水平，转染Derlin-1能够减弱吉非替尼对肿瘤细胞增殖的抑制并且能够上调Akt的活性，表明 Derlin-1可能通过Akt途径使肺癌细胞产生EGFR-TKI耐药，但Derlin-1激活Akt的分子机制及是否还通过其他途径介导耐药目前还不清楚。 本研究拟双向调控肺癌细胞中Derlin-1的表达，采用基因芯片、抗体芯片、免疫共沉淀、GST-pulldown等明确Derlin-1介导Akt通路激活的关键因子，揭示Derlin-1致耐药的机制。结合动物实验和临床肺癌标本揭示Derlin-1表达对EGFR-TKI靶向治疗敏感性的诊断预测作用。

Derlin-1 作为一个内质网应激相关蛋白在一些肿瘤中存在高表达。 我们之前的研究发现Derlin-1能够促进肺癌细胞侵袭能力。但Derlin-1过表达的机制和其对肺癌耐药的关系并不清楚。本研究发现Derlin-1高表达在EGFR突变呈现正相关。EGFR-TKI吉非替尼能通过XBP-1诱导Derlin-1上调。Derlin-1能够通过激活AKT信号通路介导肺癌细胞对吉非替尼的耐受。Derlin-1还在膀胱癌和食管癌中过表达并与肿瘤不良预后相关，Derlin-1可以通过AKT-Bcl-2信号通路介导肿瘤细胞化疗和放疗的耐受能力，减少凋亡。抑制剂阻断AKT信号通路后，Derlin-1对放化疗和靶向治疗的耐受功能明显减弱。此外，我们通过免疫共沉淀验证了Derlin-1与PI3K p110调节亚基存在着相互作用。综上，本研究证实了Derlin-1对肿瘤耐药的生物学功能和相关分子机制，揭示了Derlin-1对肿瘤放化疗和靶向治疗敏感性的诊断预测作用。为肺癌的防治提供理论基础和潜在靶点。发表相关论文6篇，其中中科院1区4篇。总影响因子达28.154。