Nigro E, Imperlini E, Scudiero O, Monaco M L, Polito R, Mazzarella G, Orrù S, Bianco A, Daniele A
CEINGE-Biotecnologie Avanzate Scarl, Via G. Salvatore 486, 80145, Naples, Italy.
IRCCS SDN, Via E. Gianturco 113, 80142, Naples, Italy.
Respir Res. 2015 Jun 24;16(1):74. doi: 10.1186/s12931-015-0234-2.
Lung cancer is a leading cause of mortality. The most common cancer subtype, non small cell lung cancer (NSCLC), accounts for 85-90% all cases and is mainly caused by environmental and genetic factors. Mechanisms involved in lung carcinogenesis include deregulation of several kinases and molecular pathways affecting cell proliferation, apoptosis and differentiation. Despite advances in lung cancer detection, diagnosis and staging, survival rate still remains poor and novel biomarkers for both diagnosis and therapy need to be identified. In the present study, we have explored the potential of novel specific biomarkers in the diagnosis of NSCLC, and the over-expression/activation of several kinases involved in disease development and progression.
Lung tumor tissue specimens and adjacent cancer-free tissues from 8 NSCLC patients undergoing surgery were collected. The differential activation status of ERK1/2, AKT and IKBα/NF-κβ was analyzed. Subsequently, protein expression profile of NSCLC vs normal surrounding tissue was compared by a proteomic approach using LC-MS MS. Subsequently, MS/MS outputs were analyzed by the Protein Discoverer platform for label-free quantitation analysis. Finally, results were confirmed by western blotting analysis.
This study confirms the involvement of ERK1/2, AKT, IKBα and NF-κβ proteins in NSCLC demonstrating a significant over-activation of all tested proteins. Furthermore, we found significant differential expression of 20 proteins (Rsc ≥ 1.50 or ≤ -1.50) of which 7 are under-expressed and 13 over-expressed in NSCLC lung tissues. Finally, we validated, by western blotting, the two most under-expressed NSCLC tissue proteins, carbonic anhydrase I and II isoforms.
Our data further support the possibility of developing both diagnostic tests and innovative targeted therapy in NSCLC. In addition to selective inhibitors of ERK1/2, AKT, IKBα and NF-κβ, as therapeutic options, our data, for the first time, indicates carbonic anhydrase I and II as attractive targets for development of diagnostic tools enabling selection of patients for a more specific therapy in NSCLC.
肺癌是主要的致死原因。最常见的癌症亚型，非小细胞肺癌（NSCLC），占所有病例的85 - 90%，主要由环境和遗传因素引起。肺癌发生所涉及的机制包括多种影响细胞增殖、凋亡和分化的激酶及分子途径的失调。尽管肺癌检测、诊断和分期取得了进展，但生存率仍然很低，需要鉴定用于诊断和治疗的新型生物标志物。在本研究中，我们探索了新型特异性生物标志物在NSCLC诊断中的潜力，以及几种参与疾病发生和进展的激酶的过表达/激活情况。
收集了8例接受手术的NSCLC患者的肺肿瘤组织标本和相邻的无癌组织。分析了ERK1/2、AKT和IKBα/NF - κβ的差异激活状态。随后，采用液相色谱 - 串联质谱（LC - MS MS）蛋白质组学方法比较NSCLC与正常周围组织的蛋白质表达谱。随后，通过Protein Discoverer平台对串联质谱输出结果进行无标记定量分析。最后，通过蛋白质印迹分析对结果进行确认。
本研究证实ERK1/2、AKT、IKBα和NF - κβ蛋白参与NSCLC，显示所有检测蛋白均有显著的过度激活。此外，我们发现20种蛋白（相对表达量变化≥1.50或≤ - 1.50）有显著差异表达，其中7种在NSCLC肺组织中低表达，13种高表达。最后，我们通过蛋白质印迹法验证了NSCLC组织中表达最低的两种蛋白，即碳酸酐酶I和II同工型。
我们的数据进一步支持了开发NSCLC诊断测试和创新靶向治疗的可能性。除了ERK1/2、AKT、IKBα和NF - κβ的选择性抑制剂作为治疗选择外，我们的数据首次表明碳酸酐酶I和II是开发诊断工具的有吸引力的靶点，能够为NSCLC患者选择更特异性的治疗。
