【作者】 王智；

【导师】 朱宇熹；

【作者基本信息】 重庆医科大学 ， 临床医学（专业学位）， 2022， 硕士

【摘要】 背景:肺癌是全世界范围内公认的高危恶性肿瘤,30-60%的非小细胞肺癌(Non-small cell lung cancer,NSCLC)患者存在表皮生长因子受体(Epidermal growth factor receptor,EGFR)基因突变使得表皮生长因子受体酪氨酸激酶抑制剂(EGFR-tyrosine kinase inhibitor,EGFR-TKI)成为了有效的治疗选择。然而患者可能对EGFR-TKI存在原发性或继发性耐药,其中涉及到的分子机制变化并未完全清楚,是急需解决的临床问题。方法:我们利用GEO数据库中对一代和二代EGFR-TKI耐药的非小细胞肺癌基因表达谱,计算得到差异表达基因。对差异表达基因进行表达水平验证、通路富集及生存分析,获取到与非小细胞肺癌预后相关的耐药关键基因。以关键基因不同表达水平及EGFR突变状态对肺腺癌进行分组,计算不同分组的免疫浸润评分,以此来分析关键基因在免疫浸润中作用。最后,我们通过实时荧光定量逆转录聚合酶链反应(Quantitative real time polymerase chain reaction,q RT-PCR)验证关键基因在临床EGFR突变非小细胞肺癌和正常癌旁组织中的表达差异,随访相应患者一线用药后的无进展生存期。结果:本研究共发现了107个与TKI耐药相关的差异表达基因。经可视化筛选后获得了11个关联紧密的耐药关键基因,其中关键基因SPP1在高表达时非小细胞肺癌患者预后明显变差。免疫浸润分析结果显示,SPP1高表达的TKI耐药组较SPP1低表达的敏感组,其CD4+T细胞的浸润评分更高,该结果可能为后续免疫治疗提供依据。最后我们通过q RT-PCR验证了SPP1在EGFR突变非小细胞肺癌较正常组织表达显著升高,且SPP1高表达患者的无进展生存期更短。结论:因此我们得出结论,SPP1的高表达可能会诱导非小细胞肺癌对一、二代EGFR-TKI耐药从而导致生存变差,并影响相关的肿瘤免疫浸润。为临床改善EGFR-TKI耐药,提高疗效提供了新的思路。更多还原

【Abstract】 Background: Lung cancer is a high-risk malignancy worldwide.The harboring of epidermal growth factor receptor(EGFR)mutations in30-60% non-small cell lung cancer(NSCLC)makes EGFR-tyrosine kinase inhibitor(EGFR-TKI)an attractive therapeutic option.However,patients usually suffer the primary and secondary resistance to EGFR-TKI.Thus the molecular alteration is still not fully clear and needs further study.Methods: The GEO database was utilized to find the differentially expressed genes(DEGs)in NSCLC resistant to the 1st and 2nd generation EGFR-TKI.We analyzed the expression and pathway enrichment of DEGs and got some hub genes.And we calculated the relationship between expression level of hub genes and prognosis of NSCLC.According to the expression level of hub gene and EGFR mutation status,lung adenocarcinoma were divided into different groups to calculate immune infiltration scores.Further,we analyzed hub gene expressions in EGFR-mutant NSCLC and paracancerous samples by q RT-PCR,and all patients were followed for progression-free survival analysis.Results: A total of 107 DEGs were found related to TKI resistance.Eleven hub genes were obtained after visualization and SPP1 was defined as prognostic hub gene.High expression of SPP1 significantly decreased the survival of NSCLC.The immune infiltration analysis showed higher CD4+ T cells infiltrated scores in the resistant group with high expression of SPP1.This result might indicate the potential of immune checkpoint inhibitors therapy after EGFR-TKI resistance.q RT-PCR validated higher expression of SPP1 in EGFR-mutant NSCLC than normal tissue,and patients with high SPP1 expression suffered from poorer progression-free survival.Conclusion: Our study suggested that the upregulation of SPP1 might induce resistance to the 1st and 2nd generation EGFR-TKI resulting in poor survival and influencing tumor immune infiltration.Therefore,these results might provide a new idea to overcome the resistance of EGFR-TKI and improve its efficacy.更多还原