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N^(6)-methyladenosine modification of CENPk mRNA by ZC3H13 promotes cervical cancer stemness and chemoresistance

Military Medical Research 2022年第5期9卷 576-591页

作者： 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 HospitalFujian Medical UniversityFuzhou 350014China Department of Radiation Oncology Fujian Cancer Hospital and Fujian Medical University Cancer HospitalFujian Medical UniversityFuzhou 350014China Shenzhen key Laboratory of Immunity and Inflammatory Diseases Peking University Shenzhen HospitalShenzhen Peking University?the Hong Kong University of Science and Technology Medical CenterShenzhen 518036GuangdongChina Outpatient Department Fujian Hospital of People's Armed PoliceFujian Medical UniversityFuzhou 350014China

Background:Stemness and chemoresistance contribute to cervical cancer recurrence and *** the current study,we determined the relevant players and role of N^(6)-methyladenine(m^(6)A)RNA methylation in cervical cancer ***:The roles of m^(6)A RNA methylation and centromere protein k(CENPk)in cervical cancer were analyzed using bioinformatics *** RNA immunoprecipitation was adopted to detect m^(6)A modification of CENPk *** cervical cancer clinical samples,cell lines,and xenografts were used for analyzing gene expression and *** 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,*** blot analysis,co-immunoprecipitation,chromatin immunoprecipitation,and luciferase reporter,cycloheximide chase,and cell fractionation assays were performed to elucidate the underlying ***:Bioinformatics analysis of public cancer datasets revealed firm links between m^(6)A modification patterns and cervical cancer prognosis,especially through ZC3H13-mediated m^(6)A modification of 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(Pk was directly bound to SOX6 and disrupted the interactions of CENPk withβ-catenin,which promotedβ-catenin expression and nuclear translocation,facilitated p53 ubiquitination,and led to activation of Wnt/β-catenin signaling,but suppression of the p53 *** dysregulation ultimately enhanced the tumorigenic pathways required for cell stemness,DNA damage repair pathways necessary f

关键词： N^(6)-methyladenosine centromere protein k Cervical cancer Stemness Chemoresistance

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Lentivirus-mediated short hairpin RNA interference of CENPk inhibits growth of colorectal cancer cells with overexpression of Cullin 4A

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World Journal of Gastroenterology 2022年第37期28卷 5420-5443页

作者： Xian Li Yi-Ru Han Xuefeng Xuefeng Yong-Xiang Ma Guo-Sheng Xing Zhi-Wen Yang Zhen Zhang Lin Shi Xin-Lin Wu Clinical Medical Research Center The Affiliated Hospital of Inner Mongolia Medical UniversityHohhot 010050Inner Mongolia Autonomous RegionChina Department of Gastrointestinal Surgery The Affiliated Hospital of Inner Mongolia Medical UniversityHohhot 010050Inner Mongolia Autonomous RegionChina Department of Pathology The Affiliated Hospital of Inner Mongolia Medical UniversityHohhot 010050Inner Mongolia Autonomous RegionChina

BACkGROUND Colorectal cancer(CRC)is one of the most common malignant tumors *** identification of novel diagnostic and prognostic biomarkers for CRC is a key research *** analysis has revealed high expression of centromere protein k(CENPk)in ***,the role of CENPk in the progression of CRC is not well *** To evaluate the effects of knockdown of CENPk and overexpression of Cullin 4A(CUL4A)in RkO and HCT116 *** Human colon cancer samples were collected and tested using a human gene expression *** identified CENPk as a potential oncogene for CRC based on bioinformatics *** vitro experiments verified the function of this *** investigated the expression of CENPk in RkO and HCT116 cells using quantitative polymerase chain reaction(qPCR),western blot,and flow *** effect of short hairpin RNA(shRNA)virus-infected RkO cells on tumor growth was evaluated in vivo using quantitative analysis of fluorescence *** evaluate the effects of knockdown of CENPk and overexpression of CUL4A in RkO and HCT116 cells,we performed a series of in vitro experiments,using qPCR,western blot,MTT assay,and flow *** We demonstrated overexpression of CENPk in human colon cancer *** was an independent risk factor in patients with *** downstream genes FBX32,CUL4A,and Yesassociated protein isoform 1 were examined to evaluate the regulatory action of CENPk in RkO *** delayed xenograft tumor emergence,slower growth rate,and lower final tumor weight and volume were observed in the CENPk short hairpin RNA virus infected group compared with the CENPk negative control *** CENPk gene interference inhibited the proliferation of RkO cells in vitro and in *** lentivirus-mediated shRNA interference of CENPk inhibited the proliferation of RkO and HCT116 colon cancer cells,with overexpression of the *** We indicated a potential role of CENPk in promo

关键词： Colorectal cancer centromere protein k Bioinformatics analysis Lentivirus-mediated short hairpin RNA interference Cullin 4A

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