Department of Clinical Biochemistry, Kermanshah University of Medical Sciences, Kermanshah, Iran.
J Genet. 2024;103.
Colorectal cancer (CRC) is known to develop due to the accumulation of both genetic and epigenetic alterations, resulting in the conversion of intestinal epithelial cells to malignant adenocarcinoma cells. Caudal type homeobox 1 () gene is a homeobox transcription factor and a selective tumour suppressor gene that is an important factor for the development of intestinal cells. This gene plays a role in the differentiation of intestinal epithelial cells, and its expression decreases in a number of cell lines derived from CRC, which suggests that a lack of expression is a risk factor for the development of colorectal carcinoma. Therefore, the methylated DNA amounts of gene in stool samples were investigated as a noninvasive method for the detection of CRC. In the present study, the methylation of gene promoter region was assessed in stool samples of 50 CRC patients and 50 healthy individuals by MethyLight PCR using two primers and a Taq Man probe, which was completely specifically designed for fully methylated DNA of the gene promoter region. The percentage of methylated reference (PMR) of the studied gene in all samples was calculated similarly to previous studies. Statistical analysis was performed using SPSS 16. The PMR medians were 3.25 (95% CI: 0.1-100) and 0.1 (95% CI: 0.07-1) in the stool samples of CRC patients and healthy individuals, respectively. The results showed a significant difference in gene PMR between stool samples of CRC patients and controls (-value\0.001). According to the results of this study, it can be argued that measurement of gene DNA in stool samples using the MethyLight PCR has acceptable sensitivity and specificity, and is adequately potential to be used as a noninvasive complementary method for the diagnosis of CRC, along with colonoscopy as the gold standard to this end. This study is the first report on methylation in stool samples of CRC patients. Therefore, further research should be carried out with a larger sample size to evaluate its efficacy as a diagnostic biomarker in clinical laboratories.
结直肠癌（CRC）的发生被认为是由于遗传和表观遗传改变的积累，导致肠上皮细胞转化为恶性腺癌细胞。尾型同源盒 1 基因（）是一种同源盒转录因子和选择性肿瘤抑制基因，是肠细胞发育的重要因素。该基因在肠上皮细胞的分化中起作用，其在源自 CRC 的许多细胞系中的表达降低，这表明 表达缺失是结直肠癌发展的危险因素。因此，研究人员通过甲基化特异性聚合酶链反应（MSP）检测了粪便样本中基因的甲基化DNA 含量，以期将其作为一种非侵入性的 CRC 检测方法。本研究采用 MethyLight PCR 检测了 50 例 CRC 患者和 50 例健康个体粪便样本中基因启动子区的甲基化情况，该方法使用两对引物和一个 Taq Man 探针，该探针是针对基因启动子区完全甲基化 DNA 而专门设计的。与以往研究类似，计算了所研究基因的甲基化参考百分比（PMR）。采用 SPSS 16 软件进行统计学分析。CRC 患者和健康个体粪便样本中基因的 PMR 中位数分别为 3.25（95%CI：0.1-100）和 0.1（95%CI：0.07-1）。CRC 患者粪便样本中基因 PMR 与对照组比较差异有统计学意义（-值\0.001）。本研究结果表明，采用 MethyLight PCR 检测粪便样本中的基因 DNA 具有较好的灵敏度和特异性，有可能成为一种非侵入性的 CRC 辅助诊断方法，与结肠镜检查作为金标准联合使用。本研究首次报道了 CRC 患者粪便样本中基因的甲基化情况。因此，应进一步扩大样本量进行研究，以评估其作为临床实验室诊断生物标志物的效能。
