Background: Stemness and chemoresistance contribute to cervical cancer recurrence and metastasis. In the current study, we determined the relevant players and role of N6-methyladenine (m6A) RNA methylation in cervical cancer progression. Methods: The roles of m6A RNA methylation and centromere protein K (CENPK) in cervical cancer were analyzed using bioinformatics analysis. Methylated RNA immunoprecipitation was adopted to detect m6A modification of CENPK mRNA. Human cervical cancer clinical samples, cell lines, and xenografts were used for analyzing gene expression and function. Immunofluorescence staining and the tumorsphere formation, clonogenic, MTT, and EdU assays were performed to determine cell stemness, chemoresistance, migration, invasion, and proliferation in HeLa and SiHa cells, respectively. Western blotting analysis, co-immunoprecipitation, chromatin immunoprecipitation, and luciferase reporter, cycloheximide chase, and cell fractionation assays were performed to elucidate the underlying mechanism. Results: Bioinformatics analysis of public cancer datasets revealed firm links between m6A modification patterns and cervical cancer prognosis, especially through ZC3H13-mediated m6A modification of CENPK mRNA. CENPK expression was elevated in cervical cancer, associated with cancer recurrence, and independently predicts poor patient prognosis [hazard ratio=1.413, 95％ confidence interval=1.078–1.853, P=0.012]. Silencing of CENPK prolonged the overall survival time of cervical cancer-bearing mice and improved the response of cervical cancer tumors to chemotherapy in vivo (P<0.001). We also showed that CENPK was directly bound to SOX6 and disrupted the interactions of CENPK with β-catenin, which promoted β-catenin expression and nuclear translocation, facilitated p53 ubiquitination, and led to activation of Wnt/β-catenin signaling, but suppression of the p53 pathway. This dysregulation ultimately enhanced the tumorigenic pathways required for cell stemness, DNA damage repair pathways necessary for cisplatin/carboplatin resistance, epithelial-mesenchymal transition involved in metastasis, and DNA replication that drove tumor cell proliferation. Conclusions: CENPK was shown to have an oncogenic role in cervical cancer and can thus serve as a prognostic indicator and novel target for cervical cancer treatment.

Xian Lin;Feng Wang;Jian Chen;Jing Liu;Yi?Bin Lin;Li Li;Chuan?Ben Chen;Qin Xu

Departments of Gynecology,Fujian Cancer Hospital and Fujian Medical University Cancer Hospital,Fujian Medical University,Fuzhou 350014,China;Department of Radiation Oncology,Fujian Cancer Hospital and Fujian Medical University Cancer Hospital,Fujian Medical University,Fuzhou 350014,China;Shenzhen Key Laboratory of Immunity and Inflammatory Diseases,Peking University Shenzhen Hospital,Shenzhen Peking University?the Hong Kong University of Science and Technology Medical Center,Shenzhen 518036,Guangdong,China;Outpatient Department,Fujian Hospital of People's Armed Police,Fujian Medical University,Fuzhou 350014,China

军事医学研究（英文）

[1]Xian Lin;Feng Wang;Jian Chen;Jing Liu;Yi?Bin Lin;Li Li;Chuan?Ben Chen;Qin Xu-.N6?methyladenosine modification of CENPK mRNA by ZC3H13 promotes cervical cancer stemness and chemoresistance)[J].军事医学研究（英文）,2022(05):576-591

CENPK,Stemness,tumorsphere,clonogenic,SOX6,carboplatin

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