Angulo-Aguado Mariana, Orjuela-Amarillo Sarah, Mora-Jácome Julián Francisco, Córdoba Lea Paloma, Gallego-Ortiz Antonio, Gaviria-Sabogal Cristian Camilo, Contreras Nora, Figueroa Carlos, Ortega-Recalde Oscar, Morel Adrien, Fonseca-Mendoza Dora Janeth
Universidad Del Rosario, School of Medicine and Health Sciences, Center for Research in Genetics and Genomics (CIGGUR), Institute of Translational Medicine (IMT), Bogotá, Colombia.
Departamento de Coloproctología, Hospital Universitario Mayor-Méderi, Universidad del Rosario, Bogotá, Colombia.
Front Med (Lausanne). 2023 Aug 3;10:1160368. doi: 10.3389/fmed.2023.1160368. eCollection 2023.
Colorectal cancer (CRC) is a prevalent cancer, ranking as the third most common. Recent advances in our understanding of the molecular causes of this disease have highlighted the crucial role of tumor immune evasion in its initiation and progression. , a receptor that acts as a negative regulator of T cell responses, plays a pivotal role in this process, and genetic variations in have been linked to CRC susceptibility, prognosis, and response to therapy.
We conducted a case-control study involving 98 CRC patients and 424 controls. We genotyped the c.-319C > T variant (rs5742909) and performed an association analysis by comparing allele frequencies between the patients and controls. To assess the potential functional impact of this variant, we first performed an analysis of transcription factor binding sites using Genomatix. Finally, to validate our findings, we conducted a luciferase reporter gene assay using different cell lines and an electrophoretic mobility shift assay (EMSA).
The case-control association analysis revealed a significant association between c.-319C > T and CRC susceptibility ( = 0.023; OR 1.89; 95% CI = 1.11-3.23). Genomatix analysis identified LEF1 and TCF7 transcription factors as specific binders to c.-319C. The reporter gene assay demonstrated notable differences in luciferase activity between the c.-319 C and T alleles in COS-7, HCT116, and Jurkat cell lines. EMSA analysis showed differences in TCF7 interaction with the C and T alleles.
c.-319C > T is associated with CRC susceptibility. Based on our functional validation results, we proposed that c.-319C > T alters gene expression at the transcriptional level, triggering a stronger negative regulation of T-cells and immune tumoral evasion.
结直肠癌（CRC）是一种常见癌症，位列第三大常见癌症。我们对该疾病分子病因的最新认识突出了肿瘤免疫逃逸在其发生和发展中的关键作用。 作为T细胞反应的负调节因子，在这一过程中起关键作用，且 的基因变异与CRC易感性、预后及治疗反应相关。
我们进行了一项病例对照研究，纳入98例CRC患者和424例对照。我们对 c.-319C>T变异（rs5742909）进行基因分型，并通过比较患者和对照之间的等位基因频率进行关联分析。为评估该变异的潜在功能影响，我们首先使用Genomatix对转录因子结合位点进行分析。最后，为验证我们的发现，我们使用不同细胞系进行荧光素酶报告基因检测和电泳迁移率变动分析（EMSA）。
病例对照关联分析显示 c.-319C>T与CRC易感性之间存在显著关联（ =0.023；OR 1.89；95%CI=1.11-3.23）。Genomatix分析确定LEF1和TCF7转录因子为 c.-319C的特异性结合因子。报告基因检测表明，在COS-7、HCT116和Jurkat细胞系中，c.-319 C和T等位基因之间的荧光素酶活性存在显著差异。EMSA分析显示TCF7与C和T等位基因的相互作用存在差异。
c.-319C>T与CRC易感性相关。基于我们的功能验证结果，我们提出 c.-319C>T在转录水平改变基因表达，引发对T细胞更强的负调节和肿瘤免疫逃逸。
