{"title":"Lycorine affects tamoxifen resistance of breast cancer via m<sup>6</sup>A-based HAGLR.","authors":"Lei Shi, Jun-Feng Jiang, Jing Zhai","doi":"10.21037/tcr-24-1077","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>N6-methyladenosine (m<sup>6</sup>A)-mediated epitranscriptomic pathway has been shown to contribute to chemoresistance and radioresistance. Our previous work confirmed the defense of lycorine against tamoxifen resistance of breast cancer (BC) through targeting HOXD antisense growth-associated long non-coding RNA (HAGLR). Whereas, the precise regulation among them remains to be elucidated. The aim of this study was to investigate the role of IGF2BP2-mediated m<sup>6</sup>A methylation in the regulation of HAGLR and its impact on lycorine's effect on tamoxifen resistance in BC.</p><p><strong>Methods: </strong>m<sup>6</sup>A status was detected via methylated RNA immunoprecipitation-quantitative polymerase chain reaction (MeRIP-qPCR). Relative expression of HAGLR and IGF2BP2 were tested by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot analysis, respectively. Cell viability, proliferation and apoptosis were estimated via Cell Counting Kit-8 (CCK-8), colony formation and flow cytometer analysis. Interplay among IGF2BP2 and HAGLR was tested by RNA immunoprecipitation (RIP) assay. IC<sub>50</sub> value of BC cells to tamoxifen was determined by 2,5-diphenyl-2H-tetrazolium bromide (MTT) assay.</p><p><strong>Results: </strong>Total m<sup>6</sup>A level in tamoxifen-resistant BC cells (TAMR/MCF-7 and TAMR/T47D) was elevated relative to corresponding parental cells and normal mammary epithelial cell line, MCF10A, either with the presence of m<sup>6</sup>A modifications within HAGLR sequence. Moreover, IGF2BP2-mediated m<sup>6</sup>A methylation drove the upregulation and stability of HAGLR in TAMR BC cells. IGF2BP2 served as a key downstream target mediating the anti-tumors of lycorine on TAMR BC. Knockdown of IGF2BP2 or HAGLR could reduce the IC<sub>50</sub> value of TAMR/MCF-7 and TAMR/T47D cells to tamoxifen.</p><p><strong>Conclusions: </strong>Our results demonstrated that lycorine inhibits tamoxifen-resistant BC by repressing IGF2BP2-mediated m<sup>6</sup>A methylation of HAGLR.</p>","PeriodicalId":23216,"journal":{"name":"Translational cancer research","volume":"13 12","pages":"6675-6687"},"PeriodicalIF":1.5000,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11730692/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Translational cancer research","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.21037/tcr-24-1077","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/12/27 0:00:00","PubModel":"Epub","JCR":"Q4","JCRName":"ONCOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
Background: N6-methyladenosine (m6A)-mediated epitranscriptomic pathway has been shown to contribute to chemoresistance and radioresistance. Our previous work confirmed the defense of lycorine against tamoxifen resistance of breast cancer (BC) through targeting HOXD antisense growth-associated long non-coding RNA (HAGLR). Whereas, the precise regulation among them remains to be elucidated. The aim of this study was to investigate the role of IGF2BP2-mediated m6A methylation in the regulation of HAGLR and its impact on lycorine's effect on tamoxifen resistance in BC.
Methods: m6A status was detected via methylated RNA immunoprecipitation-quantitative polymerase chain reaction (MeRIP-qPCR). Relative expression of HAGLR and IGF2BP2 were tested by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot analysis, respectively. Cell viability, proliferation and apoptosis were estimated via Cell Counting Kit-8 (CCK-8), colony formation and flow cytometer analysis. Interplay among IGF2BP2 and HAGLR was tested by RNA immunoprecipitation (RIP) assay. IC50 value of BC cells to tamoxifen was determined by 2,5-diphenyl-2H-tetrazolium bromide (MTT) assay.
Results: Total m6A level in tamoxifen-resistant BC cells (TAMR/MCF-7 and TAMR/T47D) was elevated relative to corresponding parental cells and normal mammary epithelial cell line, MCF10A, either with the presence of m6A modifications within HAGLR sequence. Moreover, IGF2BP2-mediated m6A methylation drove the upregulation and stability of HAGLR in TAMR BC cells. IGF2BP2 served as a key downstream target mediating the anti-tumors of lycorine on TAMR BC. Knockdown of IGF2BP2 or HAGLR could reduce the IC50 value of TAMR/MCF-7 and TAMR/T47D cells to tamoxifen.
Conclusions: Our results demonstrated that lycorine inhibits tamoxifen-resistant BC by repressing IGF2BP2-mediated m6A methylation of HAGLR.
期刊介绍：
Translational Cancer Research (Transl Cancer Res TCR; Print ISSN: 2218-676X; Online ISSN 2219-6803; http://tcr.amegroups.com/) is an Open Access, peer-reviewed journal, indexed in Science Citation Index Expanded (SCIE). TCR publishes laboratory studies of novel therapeutic interventions as well as clinical trials which evaluate new treatment paradigms for cancer; results of novel research investigations which bridge the laboratory and clinical settings including risk assessment, cellular and molecular characterization, prevention, detection, diagnosis and treatment of human cancers with the overall goal of improving the clinical care of cancer patients. The focus of TCR is original, peer-reviewed, science-based research that successfully advances clinical medicine toward the goal of improving patients'' quality of life. The editors and an international advisory group of scientists and clinician-scientists as well as other experts will hold TCR articles to the high-quality standards. We accept Original Articles as well as Review Articles, Editorials and Brief Articles.
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