Li Kexin, Pei Yuqing, Dong Xin, Wu Yue, Lou Xiaoying, Li Yiling, Liang Shuang, Wu Yuxin, Xu Danfei, Li Bin, Cui Wei
Department of Clinical Laboratory, State Key Laboratory of Molecular Oncology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
Department of Laboratory Medicine, West China Hospital of Sichuan University, Chengdu, China.
Cancer Gene Ther. 2025 May;32(5):584-594. doi: 10.1038/s41417-025-00898-5. Epub 2025 Apr 2.
N6-methyladenosine (m6A) methylation was found to be involved in the tumorigenesis and development of ovarian cancer. Until now, it is not clear to identify the mechanism by m6A demethylase ALKBH5 affects RNA splicing in ovarian cancer. In this study, we examined ALKBH5 protein expression and m6A levels by immunohistochemistry and analyzed their correlation with clinical features and prognosis in patients with ovarian cancer. We identified the elevated expression of ALKBH5 and a general reduction in the level of m6A in ovarian cancer patients. In the ovarian cancer cell line A2780, ALKBH5 depletion was found using the siRNA strategy or the CRISPR/Cas9 knockout (KO) method, which significantly reduced the level of m6A and inhibited cell viability, proliferation, and migration. The MeRIP-seq and RNA-seq showed that ALKBH5-regulated m6A RNA modification mainly affects RNA splicing function in ovarian cancer cells. SRSF10 is a key target gene involved in alternative splicing regulation through ALKBH5-m6A. ALKBH5 knockdown resulted in increased retention of SRSF10 exon 5 and decreased expression of transcript SRSF10-211. In conclusion, the alternative splicing regulation effect by ALKBH5-mediated m6A suggests a novel promising approach for m6A modification in OC and provides novel insights into the mechanisms involved in ovarian cancer therapy.
N6-甲基腺苷（m6A）甲基化被发现参与卵巢癌的发生和发展。到目前为止，尚不清楚m6A去甲基化酶ALKBH5影响卵巢癌RNA剪接的机制。在本研究中，我们通过免疫组织化学检测了ALKBH5蛋白表达和m6A水平，并分析了它们与卵巢癌患者临床特征和预后的相关性。我们发现卵巢癌患者中ALKBH5表达升高，m6A水平普遍降低。在卵巢癌细胞系A2780中，使用siRNA策略或CRISPR/Cas9基因敲除（KO）方法发现ALKBH5缺失，这显著降低了m6A水平，并抑制了细胞活力、增殖和迁移。MeRIP-seq和RNA-seq表明，ALKBH5调节的m6A RNA修饰主要影响卵巢癌细胞中的RNA剪接功能。SRSF10是通过ALKBH5-m6A参与可变剪接调控的关键靶基因。ALKBH5敲低导致SRSF10外显子5保留增加，转录本SRSF10-211表达降低。总之，ALKBH5介导的m6A对可变剪接的调控作用为卵巢癌中m6A修饰提供了一种新的有前景的方法，并为卵巢癌治疗涉及的机制提供了新的见解。
