A2BP1的表观沉默通过上调SNAI1的m6A甲基化促进结直肠癌侵袭转移的机制研究

The transcription factor SNAI1 plays a key role in tumor invasion and metastasis. In our previous study, MeRIP-seq and methylated microarrays data showed that both SNAI1 RNA methylation and A2BP1 promoter methylation levels were increased in colorectal cancer. Moreover, we found that A2BP1 promoter methylation inhibited its expression, and A2BP1 silencing was related to cancer cell invasion and metastasis, clinical recurrence and metastasis, SNAI1 RNA methylation upregulation, and SNAI1 expression upregulation. Considering A2BP1 interacts with SNAI1 RNA, we thus propose a hypothesis that DNA methylation inactivates A2BP1 and inhibits its down-regulating effect on RNA methylation of SNAI1 through recruiting the demethylase ALKBH5, and thereby enhancing SNAI1 expression and promoting tumor invasion and metastasis. This project will further study the function of tumor suppressor gene A2BP1 and its interaction with ALKBH5 and SNAI1 RNA through CRISPR methylation editing, vector construction, QASM, MeRIP, and Co-IP. This study will further elucidate the upstream regulation mechanisms of SNAI1 expression and help develop novel molecular markers and target therapeutics.

转录因子SNAI1在肿瘤侵袭转移中起着关键作用。我们前期通过MeRIP-seq和甲基化芯片发现结直肠癌中SNAI1 RNA m6A甲基化和A2BP1启动子甲基化显著升高。预实验发现，A2BP1启动子甲基化可抑制其表达，A2BP1沉默与细胞侵袭转移、临床复发转移、SNAI1 RNA m6A甲基化上调、SNAI1表达上调相关，A2BP1与SNAI1 RNA存在相互作用。因此，我们提出科学假说：DNA甲基化沉默A2BP1，抑制其募集去甲基酶ALKBH5结合SNAI1 RNA，从而上调SNAI1的RNA m6A甲基化，增强SNAI1的表达和促进结直肠癌的侵袭转移。本课题将通过CRISPR甲基化编辑、载体构建、QASM、MeRIP和Co-IP等技术进一步研究A2BP1的基因功能及其与ALKBH5和SNAI1 RNA的相互作用，探讨调控SNAI1表达的上游机制，为分子标志物和靶点药物的开发提供新思路。

结直肠癌是我国最常见的恶性肿瘤之一，是严重威胁人类健康与生命的疾病，本课题通过使用自主研发的单碱基分辨率下检测基因甲基化水平的新技术QASM、DNA甲基化芯片、RNA-seq、MeRIP、RIP和dot blot等方法，发现了一系列关键DNA甲基化和RNA甲基化位点在结直肠癌中的作用及其调控机制，并在多个医疗中心中验证了其作为肠癌标志物的应用前景、揭示了RNA m6A甲基化及其去甲基酶ALKBH5在结直肠癌发生发展中的作用，系统揭示并验证了ALKBH5的作用靶点及其在结直肠癌发生发展中的作用，本项目发现的关键基因甲基化具有预测预后的临床标志物价值，可作为新的肿瘤分子标志物和治疗靶点应用于临床，为以后设计合理的治疗药物及判断预后，实现我国结直肠癌的精准医疗具有重要意义。

内容获取失败，请点击重试