Department of Surgery, School of Medicine, Qazvin University of Medical Sciences, Qazvin, Iran.
Department of Hematology and Medical Oncology, School of Medicine, Qazvin University of Medical Sciences, Qazvin, Iran.
J Clin Lab Anal. 2022 Jan;36(1):e24147. doi: 10.1002/jcla.24147. Epub 2021 Dec 3.
Nowadays, non-invasive and rapid detection of cancers through molecular biomarkers has received much attention. Therefore, this study investigated the non-invasive and rapid diagnosis of colorectal cancer through one of the newest biomarkers (circular RNA).
For this purpose, we collected tumoral, adjacent normal tissue, and plasma samples from 100 colorectal cancer (CRC) patients, 25 postoperative CRC patients, 28 colitis patients, and 108 healthy donors. First Illumina high-throughput (Hi Seq 2000) sequencing was performed to identify known and novel differentially expressed circRNAs in the cancerous and adjacent normal tissues (n = 3). We used quantitative real-time fluorescent polymerase chain reaction (qRT-PCR) to detect the expression level of hsa_circ_0006282 among the different samples. Moreover, inter- and intra-assays were performed to evaluate the potential of hsa_circ_0006282 as being a biomarker. The receiver operating characteristic curve (ROC) was drawn to appraise its diagnostic efficacy, and the sensitivity of this circ RNA was evaluated.
Based on RNA-sequencing results circ_0006282, cirs7, circ-0001313, circ_0055625, circ_000984, circ_0055625, circ_0001178, circ_0071589, circ-001569 were upregulated, and circ-ITGA7, circ-CDYL, circITCH, circ_0026344, circ_0000038, circ_0002220, circ_0067480, circIGHV3-20-1, circ_104916, circ_0009361 were downregulated circRNA. The hsa_circ_0006282 was the highest upregulated differentially expressed circRNA. Expression evaluation of this circRNA on different samples showed upregulation in CRC tissues (p < 0.0001) and plasma samples of CRC patients in comparison to healthy controls (p < 0.0001), while the area under the curve (AUC) was 0.831 (95% CI: 0.779-0.883). Expression of hsa_circ_0006282 in CRC patients decreased to normal after surgery (p < 0.0001). Our results showed high specificity and sensitivity of CRC detection when hsa_circ_0006282, carcinoembryonic antigen (CEA), and carbohydrate antigen199 (CA199) are combined.
Plasma hsa_circ_0006282 can be used as a novel diagnostic and dynamic monitoring biomarker in CRC patients.
如今，通过分子生物标志物对癌症进行非侵入性和快速检测已受到广泛关注。因此，本研究通过最新的生物标志物之一（环状 RNA）来研究结直肠癌的非侵入性和快速诊断。
为此，我们收集了 100 例结直肠癌（CRC）患者、25 例术后 CRC 患者、28 例结肠炎患者和 108 例健康供体的肿瘤、相邻正常组织和血浆样本。首先进行 Illumina 高通量（Hi Seq 2000）测序，以鉴定癌症组织和相邻正常组织中已知和新型差异表达的 circRNAs（n=3）。我们使用实时荧光定量聚合酶链反应（qRT-PCR）检测不同样本中 hsa_circ_0006282 的表达水平。此外，进行了内部和外部测定，以评估 hsa_circ_0006282 作为生物标志物的潜力。绘制了受试者工作特征曲线（ROC）以评估其诊断效果，并评估了该 circRNA 的灵敏度。
基于 RNA-seq 结果，circ_0006282、cirs7、circ-0001313、circ_0055625、circ_000984、circ_0055625、circ_0001178、circ_0071589、circ-001569 上调，circ-ITGA7、circ-CDYL、circITCH、circ_0026344、circ_0000038、circ_0002220、circ_0067480、circIGHV3-20-1、circ_104916、circ_0009361 下调 circRNA。hsa_circ_0006282 是上调差异表达 circRNA 中最高的。对不同样本中该 circRNA 的表达评估表明，CRC 组织中存在上调（p<0.0001），CRC 患者血浆样本与健康对照相比也存在上调（p<0.0001），曲线下面积（AUC）为 0.831（95%CI：0.779-0.883）。CRC 患者手术后 hsa_circ_0006282 的表达下降至正常（p<0.0001）。当 hsa_circ_0006282、癌胚抗原（CEA）和碳水化合物抗原 199（CA199）联合使用时，检测 CRC 的特异性和灵敏度较高。
血浆 hsa_circ_0006282 可作为 CRC 患者的新型诊断和动态监测生物标志物。
