Abstract
Abstract Background: Ctherapy resistance has been a major barrier against successful theratment. Exosome (EV), an important intercellular communication mediator, participates in a highly and complex network that drug resistance of cancer cells. EV has been reported to be able to transfer drug resistance from drug-resistant tumor cells to adjacent drug-sensitive cells. We identified a novel target, UBE2R2, based on our previous exosome proteomics analysis of resistant colorectal cancer cell lines. The main goal of this study is to validate the underlying mechanisms leading to chemotherapy resistance. Methods: We isolated EV from drug resistant cancer cell lines including coloractal cancer cell lines (HCT116-OXAR, DLD-1 OXA-R, DLD-1-OXA-R/SN38) its parental cell lines (HCT116, DLD-1) using the ExoLutE isolation kits based on a combination of multiple isolation techniques. Based on the existing data (proteomic analysis), we conducted experiments to elucidate the functions and mechanisms involved in cancer resistance of the identified UBE2R2. Cell viability assay and invasion asaay were performed to assess the effects of EVs drived from drug resistance colorectal cells on oxaliplatin and irinotecan resistance. In addition, cells transfected with siControl and siUBE2R2 were treated with oxaliplatin to access the relative UBE2R2 drug resistance effect. To explore the related mechanisms, we performed real time PCR and western blot to identify the UBE2R2-SMYD3-P53 pathway, and FACS analysis to confirm apoptosis. Additionally, TMA samples were utilized to confirm resistance according to patient stage, and UBE2R2 expression was validated by immunohistological methods. Results: UBE2R2, which is involved in ubiquitination, was most prominently expressed in resistant cell lines compared to controls in our previous proteomic analysis. Based on these results, we found that cell lines treated with exosomes which were derived from resistant cell lines showed lower sensitivity to the drug. Notably, treatment of resistant cell lines with siUBE2R2 resulted in increased sensitivity, confirming the involvement of UBE2R2 in resistance. As a possible mechanism in which UBE2R2 may be involved, western analysis targeting the SMYD-P53 mechanism showed that the combination of UBE2R2-SMYD3-p53 promoted the ubiquitination and degradation of p53. Analyzed using TMA patient samples, we found that the expression of UBE2R2 increased towards more advenced stage. Conclusion: Our results suggest a novel mechanism of chemotherapy resistance may be associated with UBE2R2. Exosome (including UBE2R2) which was drived from drug resistance cells may induce oxaliplatin and irinotecan resistance in parental colorectal cells via regulation of SMYD3-UBE2R2-P53 signaling pathway, which will be useful for investigating a potential clinical target in prediction drug resistance. Citation Format: Chi Hoon Maeng, Minji Choi. The role of exosomal UBE2R2 in colorectal cancer drug resistance [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 306.
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