来自鼻咽癌福尔马林固定石蜡包埋样本的无标记定量质谱分析：欧洲非流行地区患者队列的初步结果
Label-free quantitative mass spectrometry from formalin-fixed paraffin-embedded samples of nasopharyngeal carcinoma: Preliminary results from a non-endemic European cohort of patients.
作者信息
Netto Eduardo, Santos Hugo, Carvalho Luís, Capelo-Martínez José Luis, Rito Miguel, Cabeçadas José, Roldão Margarida
机构信息
Serviço de Radioterapia, Instituto Português de Oncologia Francisco Gentil, Lisboa, Portugal.
NOVA Medical School, Universidade NOVA de Lisboa, Lisboa, Portugal.
出版信息
Rep Pract Oncol Radiother. 2020 Sep-Oct;25(5):746-753. doi: 10.1016/j.rpor.2020.05.007. Epub 2020 Jul 11.
AIM
Report our results of biomarker discovery in formalin-fixed paraffin-embedded (FFPE) nasopharyngeal carcinoma (NPC) via proteomic analysis.
BACKGROUND
Nasopharyngeal carcinoma (NPC) is a rare cancer in Western countries. Proteomic analysis have already been reported as a useful tool to provide biomarkers. Formalin-fixed paraffin-embedded (FFPE) samples, despite largely underused, can provide invaluable information for biomarker research via proteomic analysis.
METHODS
FFPE samples of NPC were submitted to protein extraction followed by FASP-digestion and label-free quantitative mass spectrometry (MS). Patients' received concurrent chemoradiation with or without adjuvant chemotherapy as per Intergroup 0099 trial. IMRT was delivered following the RTOG0615 specifications. Toxicity was scored using the CTCAE 4.03 tables. Survival was estimated using Kaplan-Meier curves. Log-rank was used to detect differences. KEGG ontology graphics were generated.
RESULTS
28 FFPE samples from NPC patients were used. Patients were: 79% male, 97% Caucasians, 86% WHO type 3, 40% T1, 10% T2, 25% T3, and 25% T4. With a median follow up of 37 months, local control was 83 (T1, 100% T2, T3 and T4), overall survival was 84%, and six patients developed distant metastases. All five patients that died were due to metastatic disease. Tumor samples contained a median of 75% of tumor material. We found Epstein-Barr (EBV) and Herpes simplex (HSV) viruses' related proteins significantly present in early-stage primary NPC (T1 and T2, < 0.01). A pool of 10 proteins was statistically up-regulated in the metastatic group of patients ( < 0.01). Median survival from this M1 group was <1 year ( < 0.001).
CONCLUSIONS
FFPE samples yielded adequate material for MS analysis. We found EBV and HSV related proteins on early-stage NPC, and proteomic profiling associated with distant metastases, potential candidates of disease biomarkers. Validation is needed.
目的
报告我们通过蛋白质组学分析在福尔马林固定石蜡包埋（FFPE）鼻咽癌（NPC）中发现生物标志物的结果。
背景
鼻咽癌（NPC）在西方国家是一种罕见癌症。蛋白质组学分析已被报道为一种提供生物标志物的有用工具。福尔马林固定石蜡包埋（FFPE）样本尽管在很大程度上未得到充分利用，但通过蛋白质组学分析可为生物标志物研究提供宝贵信息。
方法
将NPC的FFPE样本进行蛋白质提取，随后进行FASP酶解和无标记定量质谱分析（MS）。患者根据0099组间试验接受同步放化疗，有或无辅助化疗。调强放射治疗（IMRT）按照RTOG0615规范进行。使用CTCAE 4.03表格对毒性进行评分。使用Kaplan-Meier曲线估计生存率。采用对数秩检验来检测差异。生成KEGG本体图。
结果
使用了28例NPC患者的FFPE样本。患者情况为：79%为男性，97%为白种人，86%为WHO 3型，40%为T1期，10%为T2期，25%为T3期，25%为T4期。中位随访37个月，局部控制率为83%（T1期为100%，T2、T3和T4期），总生存率为84%，6例患者发生远处转移。所有5例死亡患者均死于转移性疾病。肿瘤样本中肿瘤组织的中位数为75%。我们发现爱泼斯坦-巴尔病毒（EBV）和单纯疱疹病毒（HSV）相关蛋白在早期原发性NPC（T1和T2期，P<0.01）中显著存在。一组10种蛋白质在转移患者组中统计学上调（P<0.01）。该M1组的中位生存期<1年（P<0.001）。
结论
FFPE样本为MS分析提供了足够的材料。我们在早期NPC中发现了EBV和HSV相关蛋白，以及与远处转移相关的蛋白质组学特征，这些是疾病生物标志物的潜在候选物。需要进行验证。
Zotero 插件