Qiao Dan, Gu Chenzheng, Wang Weiwei, Yan Wenhui, Jiang Chenfei, Hu Jingwen, Shang Anquan, Guo Jian
Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
Department of Laboratory Medicine, Shanghai Tongji Hospital, School of Medicine, Tongji University, Shanghai 200065, China.
J Oncol. 2022 May 11;2022:6990955. doi: 10.1155/2022/6990955. eCollection 2022.
Exosomal microRNAs (miRNAs) have been linked to the genesis and progression of certain cancers. The role and regulation mechanism of cancer-derived exosomal miRNAs in CRC, however, remain unknown.
To address this, we first used miRNA sequencing to describe the miRNA profiles of circulating exosomes in order to identify miRNAs that were differently expressed between patients with CRC and healthy controls. Transmission electron microscopy, nanoparticle tracking analysis (NTA), and western blot were used to analyze exosomes generated from CRC cells. CCK-8, wound healing, and Transwell tests were used to see whether exosomes affected CRC cell proliferation, metastasis, and apoptosis, respectively. We chose and identified hsa-miR-3937, which was abundant in tumor-generated exosomes, based on earlier RNA sequencing data of exosomes obtained and extracted from seven matched specimens of tumor tissues and surrounding normal tissues of CRC patients.
The role of hsa-miR-3937 in CRC cells was found, and silencing of hsa-miR-3937 decreased CRC cell invasion and migration in a Transwell experiment. Furthermore, we discovered that there was no link between hsa-miR-3937 expression and CRC cell apoptosis. Initially, it was discovered that BCL2L12 was the target gene of hsa-miR-3937, and the TCGA database highlighted the potential therapeutic relevance of BCL2L12. Furthermore, to identify hsa-miR-3937 as a biomarker of CRC, we used peripheral blood samples rather than patient tissues and extracted exosomes from plasma samples. To assess the expression levels and predictive usefulness of plasma exosomal hsa-miR-3937 in CRC, we performed RT-qPCR to identify hsa-miR-3937 levels in all samples. We also gathered clinicopathological information in order to look for links between aberrant hsa-miR-3937 expression and clinicopathological characteristics. The pathologic stage of CRC patients was linked to hsa-miR-3937 expression levels, and the same was true for the T stage. ROC curve study revealed that hsa-miR-3937 outperforms CEA and CA199. The combination of hsa-miR-3937, CEA, and CA199 exhibited the highest sensitivity for CRC diagnosis.
Our findings show that the tumor-originated exosomal hsa-miR-3937 is a potential and effective liquid biopsy marker for colorectal cancer detection and therapy.
外泌体微小RNA（miRNA）与某些癌症的发生和进展有关。然而，癌症来源的外泌体miRNA在结直肠癌中的作用和调控机制仍不清楚。
为了解决这个问题，我们首先使用miRNA测序来描述循环外泌体的miRNA谱，以鉴定结直肠癌患者和健康对照之间差异表达的miRNA。透射电子显微镜、纳米颗粒跟踪分析（NTA）和蛋白质印迹法用于分析结直肠癌细胞产生的外泌体。分别使用CCK-8、伤口愈合试验和Transwell试验来观察外泌体是否影响结直肠癌细胞的增殖、转移和凋亡。基于从7例结直肠癌患者肿瘤组织和周围正常组织的匹配标本中获得并提取的外泌体的早期RNA测序数据，我们选择并鉴定了在肿瘤产生的外泌体中丰富的hsa-miR-3937。
发现hsa-miR-3937在结直肠癌细胞中的作用，在Transwell实验中沉默hsa-miR-3937可降低结直肠癌细胞的侵袭和迁移。此外，我们发现hsa-miR-3937表达与结直肠癌细胞凋亡之间没有关联。最初，发现BCL2L12是hsa-miR-3937的靶基因，TCGA数据库突出了BCL2L12的潜在治疗相关性。此外，为了将hsa-miR-3937鉴定为结直肠癌的生物标志物，我们使用外周血样本而非患者组织，并从血浆样本中提取外泌体。为了评估血浆外泌体hsa-miR-3937在结直肠癌中的表达水平和预测效用，我们进行了RT-qPCR以鉴定所有样本中的hsa-miR-3937水平。我们还收集了临床病理信息，以寻找hsa-miR-3937异常表达与临床病理特征之间的联系。结直肠癌患者的病理分期与hsa-miR-3937表达水平相关，T分期也是如此。ROC曲线研究表明，hsa-miR-3937优于CEA和CA199。hsa-miR-3937、CEA和CA199的组合对结直肠癌诊断表现出最高的敏感性。
我们的研究结果表明，肿瘤来源的外泌体hsa-miR-3937是用于结直肠癌检测和治疗的潜在有效液体活检标志物。
