Bagryantsev M V, Yanyshev A A, Ryabkov M G, Abelevich A I, Dezortsev I L, Bazayev A V
MD, PhD, Surgeon; Nizhny Novgorod Regional Clinical Hospital named after N.A. Semashko, 190 Rodionova St., Nizhny Novgorod, 603126, Russia.
MD, PhD, Tutor, Department of General, Operative Surgery and Topographic Anatomy; Privolzhsky Research Medical University, 10/1 Minin and Pozharsky Square, Nizhny Novgorod, 603005, Russia; Surgeon; Nizhny Novgorod Regional Clinical Hospital named after N.A. Semashko, 190 Rodionova St., Nizhny Novgorod, 603126, Russia.
Sovrem Tekhnologii Med. 2024;16(5):45-51. doi: 10.17691/stm2024.16.5.05. Epub 2024 Oct 30.
The gold standard for colorectal cancer (CRC) diagnosis, colonoscopy with biopsy, is an invasive technique and has some limitations, while the known non-invasive methods do not possess sufficient sensitivity and specificity. The application of microRNA as a diagnostic and prognostic CRC biomarker may compensate for the colonoscopy limitations. However, there are no data in the literature on the existence of real test systems based on the evaluation of microRNA expression. Our pilot study is the first step to creating a test system for CRC diagnosis based on the analysis of microRNA expression in the tissue of the intact colon. is to assess the prospects of using the level of microRNA expression as a supplemental method for diagnosing colorectal cancer and adenomatous polyps.
Patients participating in the study were divided into three groups: group 1 included patients with CRC (n=5), group 2 - patients with polyps in the colon (n=4), and patients without oncological pathology treated for hemorrhoidal disease without exacerbation (n=5) composed group 3.Tissue samples of the intact gut were taken from all patients. In groups 1 and 2, biopsy was performed in the process of right-sided laparoscopic resection of the colon with tumor. Samples of the mucous membrane from the distal part of the rectum in patients of group 3 were also collected intraoperatively; they were operated on using Milligan-Morgan technique. In groups 1 and 2, CRC and polyp samples, respectively, were taken for the analysis additionally to the intact gut area.The test panel included the following microRNAs: hsa-miR-10b-5p, hsa-miR-20a-5p, hsa-miR-141-3p, hsa-miR-181b-5p. The levels of the reference genes were analyzed with the help of real-time polymerase chain reaction using intercalating SYBR Green stain.
Expression of hsa-miR-141-3p in the mucous membrane of the colon in patients of groups 1 and 2 (with CRC and polyps, respectively) was statistically significantly higher than in patients without bowel tumors. At the same time, the expression level of hsa-miR-10b-5p was statistically significantly lower in the tumor tissue (cancer or polyps) in comparison with patients of group 3.Lower values of expression in all tested microRNAs have been detected in the CRC tissue relative to the intact mucosa of the same patients. A similar tendency was also observed in patients with adenomatous polyps.
The results of the study have shown that of four microRNAs, included into the test panel, hsa-miR-141-3p and hsa-miR-10b-5p were found to have the diagnostic value for identifying tumor colorectal lesions. Thus, our data will assume that supplementing the endoscopic tests of the large intestine by the epigenetic analysis of the mucous membrane is a promising approach to cancer screening procedures.
结直肠癌(CRC)诊断的金标准——结肠镜检查并活检,是一种侵入性技术且存在一些局限性,而已知的非侵入性方法又缺乏足够的敏感性和特异性。将微小RNA用作CRC的诊断和预后生物标志物或许可以弥补结肠镜检查的局限性。然而,文献中尚无基于微小RNA表达评估的实际检测系统的相关数据。我们的初步研究是基于对完整结肠组织中微小RNA表达的分析来创建CRC诊断检测系统的第一步,目的是评估将微小RNA表达水平用作诊断结直肠癌和腺瘤性息肉补充方法的前景。
参与研究的患者分为三组:第1组包括CRC患者(n = 5),第2组为结肠息肉患者(n = 4),第3组由无肿瘤病理学病变且因痔疮疾病接受治疗且无病情加重的患者组成(n = 5)。从所有患者身上采集完整肠道的组织样本。在第1组和第2组中,在右侧腹腔镜下切除结肠肿瘤的过程中进行活检。第3组患者在术中还收集了直肠远端黏膜样本;他们采用Milligan - Morgan技术进行手术。在第1组和第2组中,除了完整肠道区域外,分别采集了CRC和息肉样本用于分析。检测面板包括以下微小RNA:hsa - miR - 10b - 5p、hsa - miR - 20a - 5p、hsa - miR - 141 - 3p、hsa - miR - 181b - 5p。借助使用嵌入型SYBR Green染料的实时聚合酶链反应分析参考基因的水平。
第1组和第2组(分别患有CRC和息肉)患者结肠黏膜中hsa - miR - 141 - 3p的表达在统计学上显著高于无肠道肿瘤的患者。同时,与第3组患者相比,肿瘤组织(癌症或息肉)中hsa - miR - 10b - 5p的表达水平在统计学上显著降低。在CRC组织中检测到的所有测试微小RNA的表达值相对于同一患者的完整黏膜均较低。在腺瘤性息肉患者中也观察到了类似趋势。
研究结果表明,在检测面板所包含的四种微小RNA中,hsa - miR - 141 - 3p和hsa - miR - 10b - 5p对识别结直肠肿瘤病变具有诊断价值。因此,我们的数据表明通过对黏膜进行表观遗传学分析来补充大肠内镜检查是癌症筛查程序的一种有前景的方法。