Abstract
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.
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Fig. 1: High expression of ALKBH5 could predict the poor survival of ovarian cancer patients.
Fig. 2: ALKBH5 knockdown inhibits cell proliferation, cell cycle, and migration of ovarian cancer cells.
Fig. 3: RNA splicing pathway for the major enrichment of m6A-modified genes in ovarian cancer cells.
Fig. 4: RNA splicing pathway for the major enrichment of m6A-modified genes in ovarian cancer cells.
Fig. 5: SRSF10 is a key target gene for alternative splicing regulation exerted by ALKBH5-m6A.
Data availability
The datasets during and/or analyzed during the current study available from the corresponding author on reasonable request.
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Funding
This study was supported by the National Natural Science Foundation of China (No. 82072361 and No. 82302625) and the Natural Science Foundation of Sichuan Province (Grant No. 2023NSFSC1485) and the Chinese Academy of Medical Sciences Innovation Fund for Medical Sciences (2023-I2M-2-004).
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These authors contributed equally: Kexin Li, Yuqing Pei, Xin Dong.
Authors and Affiliations
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
Kexin Li, Xin Dong, Yue Wu, Xiaoying Lou, Yiling Li, Shuang Liang, Yuxin Wu & Danfei Xu
Department of Laboratory Medicine, West China Hospital of Sichuan University, Chengdu, China
Yuqing Pei
Sichuan Clinical Research Center for Laboratory Medicine, Chengdu, China
Yuqing Pei
Clinical Laboratory Medicine Research Center of West China Hospital, Chengdu, China
Yuqing Pei
Department of Gynecology Oncology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
Bin Li
Department of Clinical Laboratory, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Key Practice of Laboratory Medicine in Qinghai Province, Beijing, China
Wei Cui
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Kexin Li
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Contributions
KL was responsible for the conception, methodology, software, data curation, writing, reviewing, and editing the manuscript. YP and XD was responsible for methodology, writing, and validating the study. YW, XL, YL, SL, YW, and DX were responsible for writing, editing, and validating the study. BL contributed to the sample collection and supervision the study. WC and BL contributed to the conception, resources, supervision, and funding acquisition of the study. All the authors have read the final version of the manuscript and approved its submission.
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Correspondence to Bin Li or Wei Cui.
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The authors declare no competing interests.
Ethics approval and consent to participate
All methods were performed in accordance with the relevant guidelines and regulations. Clinical studies were approved by the Ethics Committee of the Cancer Hospital of the Chinese Academy of Medical Sciences and Peking Union Medical College (approval number: NCC2020C-126; Beijing, China). All animal experiments were conducted in accordance with the guidelines of the Animal Research Ethics Committee of Cancer Hospital of the Chinese Academy of Medical Sciences and Peking Union Medical College (approval number: UJS-NCC2020A062; Beijing, China). Informed consent for participation in this study was obtained from the patients with OC.
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Li, K., Pei, Y., Dong, X. et al. ALKBH5-mediated m6A regulates the alternative splicing events of SRSF10 in ovarian cancer. Cancer Gene Ther (2025). https://doi.org/10.1038/s41417-025-00898-5
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Received:29 June 2024
Revised:23 January 2025
Accepted:21 March 2025
Published:02 April 2025
DOI:https://doi.org/10.1038/s41417-025-00898-5
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