胃癌相关血浆circRNA标志物的筛选及RT-ddPCR同时定量检测血浆hsa_circ_0001017和hsa_circ_0061276方法的建立

环状RNA（circRNA）在肿瘤发生发展过程中起重要作用，而它们的结构高稳定特性使其在作为新型肿瘤标志物方面呈现优势。课题组最近筛选出一些胃癌相关circRNA，并经细胞实验发现hsa_circ_0001017和hsa_circ_0061276调控胃癌发生。由于血浆是最常用的临床检验标本，而逆转录-微滴数字PCR（RT-ddPCR）技术具有灵敏度高、干扰少、绝对定量等优点，特别适用于检测血浆RNA类标志物。为此，本课题将优化扩增片段大小和引物浓度，建立RT-ddPCR同时定量检测血浆hsa_circ_0001017和hsa_circ_0061276的方法；分析影响血浆circRNA稳定性的因素；比较健康人、良性胃病、胃癌前病变、早期胃癌和晚期胃癌等5组人血浆hsa_circ_0001017和hsa_circ_0061276拷贝数的变化规律，分析其诊断价值，希望提高胃癌的非创性诊断水平。

Circular RNAs (circRNAs) play important roles in the occurrence and development of tumors. The property of structural stable makes circRNAs advantage as a new type of tumor markers. Recently, our group screened some gastric cancer-associated circRNAs and found that hsa_circ_0001017 and hsa_circ_0061276 regulated gastric cancer occurrence. Since that plasma is the most commonly used sample in laboratory medicine; and that reverse transcription-droplet digital polymerase chain reaction (RT-ddPCR) is more suitably used in the detection of plasma RNA-typed markers for its advantages such as high sensitivity, less interference, and absolutely quantitative detection, in this study, we will optimize the size of amplified fragments and primers' concentrations; establish a simultaneously quantitative method of plasma hsa_circ_0001017 and hsa_circ_0061276; analyze the factors that affect the stability of plasma circRNAs; compare the copy changes of plasma hsa_circ_0001017 and hsa_circ_0061276 among health controls, patients with benign gastric cancer, gastric precancerous lesion, early gastric cancer and advantaged gastric cancer; and finally analyze their diagnostic values. The study may improve the non-invasive diagnostic levels of gastric cancer.