Abstract：
RBM15 functions as an oncogene in multi-type cancers. However, the reports on the roles of RBM15 in cervical cancer are limited. The purpose of this study was to investigate the potentials of RBM15 in cervical cancer. RT-qPCR was conducted to determine mRNA levels. Western was carried out to detect protein expression. CCK-8, colony formation and EdU assays were conducted to determine cell proliferation. Scratch and transwell assays were conducted to determine cell migration and invasion. MeRIP assay was conducted to determine N6-methyl adenosine (m6A) levels. Luciferase assay was conducted to verify the m6A sites of EZH2 and binding sites between EZH2 and promoter of FN1. ChIP assay was conducted to verify the interaction between EZH2 and FN1. The results showed that RBM15 was upregulated in cervical cancer patients and cells. Moreover, high levels of RBM15 predicted poor clinical outcomes. RBM15 knockdown inhibited the proliferation and epithelial-mesenchymal transition (EMT) of cervical cancer cells. RBM15 promoted the m6A modification of EZH2 as well as its protein translation. Additionally, EZH2 bound to the promoter of fibronectin 1 (FN1) and EZH2-FN1 axis is the cascade downstream of RBM15. Overexpressed EZH2 antagonized the effects of RBM15 knockdown and promoted the aggressiveness of cervical cancer cells. In summary, RBM15/EZH2/FN1 signaling cascade induces the proliferation and EMT of cervical cancer. Therefore, RBM15/EZH2/FN1 signaling may be a promising strategy for cervical cancer.
摘要：
RBM15在多类型癌症中作为癌基因发挥作用。然而,关于RBM15在宫颈癌中的作用的报道有限.目的探讨RBM15在宫颈癌中的应用潜力。进行RT-qPCR以确定mRNA水平。进行Western检测蛋白表达。进行CCK-8、集落形成和EdU测定以确定细胞增殖。进行划痕和transwell测定以确定细胞迁移和侵袭。进行MeRIP测定以确定N6-甲基腺苷(m6A)水平。进行荧光素酶测定以验证EZH2的m6A位点以及EZH2与FN1的启动子之间的结合位点。进行ChIP测定以验证EZH2和FN1之间的相互作用。结果表明，RBM15在宫颈癌患者和细胞中上调。此外，高水平的RBM15预测不良的临床结局.RBM15敲低抑制宫颈癌细胞的增殖和上皮间质转化（EMT）。RBM15促进了EZH2的m6A修饰及其蛋白质翻译。此外,与纤连蛋白1(FN1)和EZH2-FN1轴的启动子结合的EZH2是RBM15下游的级联。过表达的EZH2拮抗RBM15敲低的作用并促进宫颈癌细胞的侵袭性。总之,RBM15/EZH2/FN1信号级联诱导宫颈癌的增殖和EMT。因此,RBM15/EZH2/FN1信号传导可能是宫颈癌的一种有希望的策略。
