Investigation of the Impact of m6A Methylation Modifications of YTHDF1 and WTAP Genes on Tumor Differentiation in Colorectal Cancer in Lishui City

目的：目前尚不清楚m6A甲基化基因在结直肠癌分化中的作用。本研究结合生物信息学分析与实验验证，探讨这些基因在结直肠癌中的表达特征及其潜在机制。方法：利用生物信息学，我们发现YTHDF1、YTHDF2、ALKBH5、WTAP与结直肠癌表达异常显著(p Background: The role of m6A methylation genes in the differentiation of colorectal cancer is currently unclear. This study combines bioinformatics analysis and experimental validation to explore the expression patterns of these genes in colorectal cancer and their potential mechanisms. Methods: Using bioinformatics, we found that YTHDF1, YTHDF2, ALKBH5, and WTAP are significantly aberrantly expressed in colorectal cancer (p < 0.05). In a real-world cohort from Lishui, quantitative PCR (qPCR) was used to measure mRNA expression of these four genes in CRC tissues and adjacent normal tissues. Multivariable logistic regression analysis with SPSS identified YTHDF1 and WTAP mRNA expression levels as being associated with CRC differentiation. Immunohistochemistry (IHC) was performed to examine corresponding protein levels, quantified using ImageJ. Results showed that YTHDF1 protein expression negatively correlated with CRC differentiation, whereas WTAP protein expression positively correlated. Additionally, multivariable Cox regression analysis was conducted to identify potential factors influencing CRC development. Results: In Lishui, YTHDF1 protein expression was negatively associated with CRC differentiation, while WTAP protein expression was positively associated. Age and alcohol consumption were key risk factors for CRC development, while no significant associations were found with sex or smoking. GO enrichment analysis suggests that YTHDF1 and WTAP may regulate the differentiation process of colorectal cancer by participating in mechanisms such as mRNA modification, translation regulation, and RNA stability. For the poorly differentiated CRC group, time to progression rate (TTP) was 100% and overall survival rate (OS) was 0% within three years. Conclusions: In Lishui City, the YTHDF1 and WTAP genes influence the differentiation of colorectal cancer and may serve as important therapeutic targets and biomarkers for the diagnosis of patients with colorectal cancer.