摘要
目的:肾癌早期无明显症状,且无血清诊断标志物,晚期缺乏有效治疗手段。外泌体的生物学功能及现有研究报道为早期肾癌的诊断提供了新的理论基础和新思路,本文旨在尝试在肾癌血清外泌体中寻找肾癌诊断标志物以有助于提高肾癌的诊断水平,减少肾癌死亡率。方法:取肾癌肿瘤组和对照组各10个样本,用试剂盒(PEG法)分离外泌体,提取RNA后建库测序,数据处理后以|log2(fold change)|>1和显著水平q_value<0.001为标准筛选得一组显著差异基因,设计引物探针后大样本qPCR验证,对数据进行统计并作t检验分析,通过LASSO回归建立诊断模型,ROC-AUC判断诊断价值。结果:RNA-seq并深度分析筛出PFN2RAB4B PHEX DACH1 Linc01765五个基因,运用LASSO回归建立了早期肾癌诊断模型,计算ROC-AUC值是0.818。结论:在肾癌血清外泌体中筛到一组基因,通过LASSO回归建立早期肾癌诊断模型,ROC-AUC值达0.818,具有一定的临床应用价值。
Objective:Early renal cell carcinoma(RCC)has no significant clinical manifestation,also lack of specific serum biomarkers.There is no effective treatment for advanced RCC.The biological function of exosomes and the existing research provide a new theoretical basis and new ideas for the diagnosis of early RCC.This paper attempts to find the diagnostic biomarkers in serum exosomes of RCC in order to improve the diagnostic level and reduce the mortality of RCC.Methods:The exosomes were isolated with commercial kit(PEG method)from 10 samples of renal tumor group and control group.RNA was extracted and sequenced.After data processing,the data were analyzed with|log2(fold change)>1 and significant level Q_Value<0.001 was used as the standards to screen a group of significantly different genes.Primers and probes were designed and qPCR was used for validation with large samples.The data were analyzed by t-test.The diagnostic model was established by LASSO regression,and the diagnostic value was judged by ROC-AUC.Results:Five genes(pfn2 rab4 b PHEX Dach1 linc01765)were screened by RNA-SEQ and deep analysis.The diagnostic model of early RCC was established by lasso regression.The ROC-AUC value was 0.818.Conclusion:A group of genes were screened from the serum exosomes of renal cell carcinoma,and the diagnostic model of early renal cell carcinoma was established by lasso regression.The ROC-AUC value was 0.818,which has certain clinical value.
作者
陈继胜
吴巧
杨安桥
谢伟建
卢大儒
CHEN Ji-sheng;WU Qiao;YANG An-qiao;XIE Wei-jian;LU Da-ru(School of Life Sciences,Fudan University,Shanghai,200433,China;Suzhou Suyuanjingwei Biotechnology CO.,Ltd,Suzhou,Jiangsu,215124,China)
出处
《现代生物医学进展》
CAS
2021年第12期2201-2206,共6页
Progress in Modern Biomedicine
关键词
外泌体
肿瘤标志物
早期诊断
LASSO回归
肾癌
Exosome
Tumor biomarkers
Early diagnosis
LASSO regression
Renal cell carcinoma
