血清环状RNA（circRNA）作为食管鳞癌的诊断标志物的研究

Non-coding RNA in human serum, especially circular RNA (circRNA), has been expected as a new tumor molecular markers in recent years. Our preliminary experiment with new-onset esophageal squamous cell carcinoma (ESCC) patients, which was screened by circRNAs sequencing, resulted in 23 differential expressed circRNAs being certified between preoperative and postoperative serum samples. We hypothesized that a set of serum circRNAs in ESCC patients can be identified as innovative tumor molecular markers with high sensitivity and specificity. The research project will take multi-step verification procedures，including surgical specimens verification, functional studies and larger sample validation. Firstly, specimens verification was implemented in surgical specimens to further validated 23 circRNAs screened by the preliminary experiment to find significantly differentially expressed circRNAs. And then the functional experiments were performed by cell cycle, cell proliferation, apoptosis, cell invasion and xenograft studies. Furthermore, larger sample case-control study was carried out to validate the candidate circRNAs, determining the potential diagnostic biomarkers, and establishing circRNAs database for ESCC. Our results provides reference data for the early diagnosis, antidiastole, follow-up and target therapy for ESCC, and bring forward theoretical basis for the molecular mechanisms of ESCC.

人体血清非编码RNA，特别是环状RNA（circRNA）有望成为新的肿瘤分子标志物。我们前期的实验以新发食管鳞癌（ESCC）患者的术前、术后血清作为自身对照，经circRNA测序比对得到23个显著差异表达的circRNA，在此基础上提出“循环血液中存在circRNA组合，可作为ESCC敏感性和特异性较强的新的肿瘤分子标志物”的假说。为此，本项目将采取手术标本验证、功能研究及扩大样本验证等多步验证方法，将预实验中筛选得到的circRNA在手术标本中进一步验证，找到显著差异表达的circRNA。通过细胞周期、细胞增殖、细胞凋亡、细胞侵袭和异种移植实验对差异表达的circRNA进行功能学研究，继而采用病例-对照研究对circRNA进行扩大样本验证，找到具有诊断标志物作用的circRNA，并建立ESCC的circRNA筛查数据库，为ESCC的早期诊断、鉴别诊断、分子机制研究及精准治疗提供参考依据。

食管鳞癌（ESCC）缺乏有效的诊断和预后生物标志物。人体血清环状RNA（circRNA）有望成为新的肿瘤分子标志物。本课题组原创性发现食管鳞癌患者血清及肿瘤组织中存在一个异常上调的环状RNA-circ4771(hsa_circ_0004771)，具备有诊断和预后的双重价值。表型研究发现circ4771在体内、体外均可促进食管鳞癌细胞增殖。机制方面，circ4771作为miR-339-5p的分子海绵，正向调控CDC25A，促进ESCC细胞周期信号通路加速运转，进而发挥促进ESCC进展的作用。本项目这部分的主要结果已经撰写成研究报告发表于Epigenomics杂志。另外进一步的，我们还发现circ4771可分泌进入外泌体，在血浆中以外泌体负载的形式转运，发挥对肿瘤微环境的调控作用，目前该部分实验仍然在进行中。我们初步证实了circ4771可以作为食管鳞癌的诊断及预后血清学标志物。