{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,22]],"date-time":"2026-07-22T09:36:18Z","timestamp":1784712978558,"version":"3.55.0"},"reference-count":58,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2025,2,9]],"date-time":"2025-02-09T00:00:00Z","timestamp":1739059200000},"content-version":"vor","delay-in-days":8,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Clinical &amp; Translational Med"],"published-print":{"date-parts":[[2025,2]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Acquired anlotinib resistance is still a key challenge in osteosarcoma treatment. Unravelling the mechanisms underlying anlotinib resistance is the key to optimising its efficacy for treating osteosarcoma. Previous studies have explored the pivotal function of the DNA\u2010dependent protein kinase catalytic subunit (DNA\u2010PKcs) with regard to osteosarcoma chemoresistance.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We used bioinformatics analysis to predict DNA\u2010PKcs and Beclin\u20101 interactions, confirmed through immunofluorescence (IF) and co\u2010immunoprecipitation (co\u2010IP). Dual\u2010luciferase analyses and Methylated RNA immunoprecipitation (MeRIP) were implemented to detect the detected m<jats:sup>6<\/jats:sup>A modifications. RNA fluorescence in situ hybridisation (FISH)\u2014IF co\u2010localisation and RNA immunoprecipitation (RIP) were conducted to explore the interplay between PRKDC mRNA and the indicated proteins.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Anlotinib\u2010treated osteosarcoma cells exhibited increased DNA\u2010PKcs levels, and silencing DNA\u2010PKcs augmented osteosarcoma sensitivity to anlotinib. DNA\u2010PKcs affects anlotinib\u2010induced autophagy by interacting with Beclin\u20101 and regulating its ubiquitination. Notably, PRKDC mRNA, encoding DNA\u2010PKcs, underwent N<jats:sup>6<\/jats:sup>\u2010Methyladenosine (m<jats:sup>6<\/jats:sup>A) modification. Methyltransferase\u2010like 3 (METTL3) positively regulated DNA\u2010PKcs expression. Functionally, METTL3 enhances anlotinib resistance in osteosarcoma, which is reversed by PRKDC knockdown. Mechanistically, METTL3 binds to PRKDC mRNA and facilitates m<jats:sup>6<\/jats:sup>A methylation. Additionally, m<jats:sup>6<\/jats:sup>A methylated PRKDC mRNA is identified via YTH N<jats:sup>6<\/jats:sup>\u2010methyladenosine RNA\u2010binding protein 1 (YTHDF1), augmenting its expression.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>These findings revealed that DNA\u2010PKcs promotes anlotinib resistance by regulating protective autophagy, while METTL3 induces PRKDC m<jats:sup>6<\/jats:sup>A modification, enhancing its expression. Thus, targeting METTL3\/PRKDC may be a novel strategy for improving therapeutic efficacy in human osteosarcoma.<\/jats:p><\/jats:sec><jats:sec><jats:title>Key points<\/jats:title><jats:p><jats:list list-type=\"bullet\">\n<jats:list-item><jats:p>DNA\u2010PKcs knockdown heightens osteosarcoma sensitivity to anlotinib.<\/jats:p><\/jats:list-item>\n<jats:list-item><jats:p>DNA\u2010PKcs modulates anlotinib\u2010induced protective autophagy through interacts with Beclin\u20101 and regulates its ubiquitination.<\/jats:p><\/jats:list-item>\n<jats:list-item><jats:p>m<jats:sup>6<\/jats:sup>A modification of OLE_LINK82PRKDC mRNA induced by METTL3 contributes to anlotinib resistance in osteosarcoma.<\/jats:p><\/jats:list-item>\n<jats:list-item><jats:p>m<jats:sup>6<\/jats:sup>A methylation of PRKDC mRNA recognised by YTHDF1 amplifies the expression of DNA\u2010PKcs.<\/jats:p><\/jats:list-item>\n<\/jats:list><\/jats:p><\/jats:sec>","DOI":"10.1002\/ctm2.70228","type":"journal-article","created":{"date-parts":[[2025,2,10]],"date-time":"2025-02-10T23:16:45Z","timestamp":1739229405000},"source":"Crossref","is-referenced-by-count":6,"title":["N<sup>6<\/sup>\u2010Methyladenosine modification mediated by METTL3 promotes DNA\u2010PKcs expression to induce anlotinib resistance in osteosarcoma"],"prefix":"10.1002","volume":"15","author":[{"given":"Yining","family":"Zhang","sequence":"first","affiliation":[{"name":"The First Clinical College of Cheeloo College of Medicine Shandong University  Jinan Shandong China"},{"name":"Department of Orthopaedics Qilu Hospital of Shandong University  Jinan Shandong China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guohong","family":"Shen","sequence":"additional","affiliation":[{"name":"Department of Pediatrics Central Hospital Affiliated to Shandong First Medical University  Jinan Shandong China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dan","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Cardiovascular Medicine Jinan Central Hospital  Jinan Shandong China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tingting","family":"Meng","sequence":"additional","affiliation":[{"name":"Department of Cardiovascular Medicine Jinan Central Hospital  Jinan Shandong China"},{"name":"Research Center of Translational Medicine Central Hospital Affiliated to Shandong First Medical University  Jinan Shandong China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhaorui","family":"Lv","sequence":"additional","affiliation":[{"name":"Department of Orthopedics Affiliated Hospital of Shandong University of Traditional Chinese Medicine  Jinan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lei","family":"Chen","sequence":"additional","affiliation":[{"name":"The First Clinical College of Cheeloo College of Medicine Shandong University  Jinan Shandong China"},{"name":"Department of Orthopaedics Qilu Hospital of Shandong University  Jinan Shandong China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianmin","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Orthopaedics Qilu Hospital of Shandong University  Jinan Shandong China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-9528-9864","authenticated-orcid":false,"given":"Ka","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Orthopaedics Qilu Hospital of Shandong University  Jinan Shandong China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,2,9]]},"reference":[{"key":"e_1_2_12_2_1","doi-asserted-by":"publisher","DOI":"10.1093\/annonc\/mdq276"},{"key":"e_1_2_12_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.canlet.2020.12.024"},{"key":"e_1_2_12_4_1","doi-asserted-by":"publisher","DOI":"10.1080\/14737140.2018.1413939"},{"key":"e_1_2_12_5_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms21186885"},{"key":"e_1_2_12_6_1","doi-asserted-by":"publisher","DOI":"10.1002\/ijc.32180"},{"key":"e_1_2_12_7_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13046-019-1093-3"},{"key":"e_1_2_12_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/bjc.2017.478"},{"key":"e_1_2_12_9_1","doi-asserted-by":"publisher","DOI":"10.1001\/jamaoncol.2018.3039"},{"key":"e_1_2_12_10_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13045-016-0332-8"},{"key":"e_1_2_12_11_1","doi-asserted-by":"publisher","DOI":"10.1002\/cam4.4286"},{"key":"e_1_2_12_12_1","doi-asserted-by":"publisher","DOI":"10.3389\/fonc.2023.1283932"},{"key":"e_1_2_12_13_1","doi-asserted-by":"publisher","DOI":"10.3389\/fphar.2021.798837"},{"key":"e_1_2_12_14_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41401-020-0421-7"},{"key":"e_1_2_12_15_1","doi-asserted-by":"publisher","DOI":"10.2147\/IJN.S321720"},{"key":"e_1_2_12_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.redox.2021.102076"},{"key":"e_1_2_12_17_1","doi-asserted-by":"publisher","DOI":"10.3389\/fonc.2023.1192472"},{"key":"e_1_2_12_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.canlet.2022.215660"},{"key":"e_1_2_12_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2017.05.045"},{"key":"e_1_2_12_20_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.2004880"},{"key":"e_1_2_12_21_1","doi-asserted-by":"publisher","DOI":"10.1038\/cr.2014.3"},{"key":"e_1_2_12_22_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41422-018-0040-8"},{"key":"e_1_2_12_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biopha.2020.109964"},{"key":"e_1_2_12_24_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12943-019-1036-9"},{"key":"e_1_2_12_25_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-021-02271-9"},{"key":"e_1_2_12_26_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtn.2020.01.033"},{"key":"e_1_2_12_27_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymthe.2020.06.024"},{"key":"e_1_2_12_28_1","doi-asserted-by":"publisher","DOI":"10.3892\/or.2016.4991"},{"key":"e_1_2_12_29_1","doi-asserted-by":"publisher","DOI":"10.1007\/s13277-015-3642-5"},{"key":"e_1_2_12_30_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.intimp.2023.111181"},{"key":"e_1_2_12_31_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12943-020-01281-8"},{"key":"e_1_2_12_32_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrg3724"},{"key":"e_1_2_12_33_1","doi-asserted-by":"publisher","DOI":"10.1038\/cr.2017.15"},{"key":"e_1_2_12_34_1","doi-asserted-by":"publisher","DOI":"10.1634\/theoncologist.2018-0542"},{"key":"e_1_2_12_35_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.dnarep.2016.10.004"},{"key":"e_1_2_12_36_1","doi-asserted-by":"publisher","DOI":"10.14348\/molcells.2023.2164"},{"key":"e_1_2_12_37_1","doi-asserted-by":"publisher","DOI":"10.1172\/JCI127483"},{"key":"e_1_2_12_38_1","doi-asserted-by":"publisher","DOI":"10.1186\/1476-4598-13-232"},{"key":"e_1_2_12_39_1","doi-asserted-by":"publisher","DOI":"10.1158\/1535-7163.MCT-21-0215"},{"key":"e_1_2_12_40_1","doi-asserted-by":"publisher","DOI":"10.1126\/scitranslmed.abc7275"},{"key":"e_1_2_12_41_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.canlet.2010.10.001"},{"key":"e_1_2_12_42_1","doi-asserted-by":"publisher","DOI":"10.1155\/2013\/102735"},{"key":"e_1_2_12_43_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41419-018-1054-3"},{"key":"e_1_2_12_44_1","doi-asserted-by":"publisher","DOI":"10.4161\/auto.6.7.13038"},{"key":"e_1_2_12_45_1","doi-asserted-by":"publisher","DOI":"10.1038\/cddis.2017.422"},{"key":"e_1_2_12_46_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrc.2017.53"},{"key":"e_1_2_12_47_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41419-020-2352-0"},{"key":"e_1_2_12_48_1","doi-asserted-by":"publisher","DOI":"10.1530\/ERC-23-0036"},{"key":"e_1_2_12_49_1","doi-asserted-by":"publisher","DOI":"10.1590\/1414-431x2023e13152"},{"key":"e_1_2_12_50_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bcp.2011.12.029"},{"key":"e_1_2_12_51_1","doi-asserted-by":"publisher","DOI":"10.1080\/15548627.2019.1710430"},{"key":"e_1_2_12_52_1","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-04-4202"},{"key":"e_1_2_12_53_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.brainres.2010.11.044"},{"key":"e_1_2_12_54_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ebiom.2020.102955"},{"key":"e_1_2_12_55_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2019.06.128"},{"key":"e_1_2_12_56_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41419-020-03315-x"},{"key":"e_1_2_12_57_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2015.05.014"},{"key":"e_1_2_12_58_1","doi-asserted-by":"publisher","DOI":"10.3389\/fonc.2019.00332"},{"key":"e_1_2_12_59_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature12730"}],"container-title":["Clinical and Translational Medicine"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/ctm2.70228","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,7,26]],"date-time":"2025-07-26T03:46:58Z","timestamp":1753501618000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/ctm2.70228"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,2]]},"references-count":58,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2025,2]]}},"alternative-id":["10.1002\/ctm2.70228"],"URL":"https:\/\/doi.org\/10.1002\/ctm2.70228","archive":["Portico"],"relation":{},"ISSN":["2001-1326","2001-1326"],"issn-type":[{"value":"2001-1326","type":"print"},{"value":"2001-1326","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,2]]},"article-number":"e70228"}}