Abstract
Background
Radiotherapy is widely employed in colorectal cancer (CRC) treatment but is often compromised by developed radioresistance. This study explored the mechanism of long non-coding RNA ovarian tumor domain containing 6B-antisense RNA1 (lncRNA OTUD6B-AS1) in CRC radioresistance through tripartite motif 16 (TRIM16).
Methods
CRC and non-cancerous tissues were collected and radioresistant CRC cells were established, with real-time quantitative polymerase chain reaction to determine gene expression in tissues and cells. Radioresistance was evaluated by cell counting kit-8 assay and immunofluorescence (γ-H2AX) and ferroptosis was tested by Western blot assay (ACSL4/GPX4) and assay kits (Fe2+/ROS/MDA/GSH). The association between ferroptosis and lncRNA OTUD6B-AS1-inhibited radioresistance was testified using ferroptosis inhibitor. The subcellular localization of lncRNA OTUD6B-AS1 was tested by the nuclear/cytoplasmic fractionation assay, with RNA immunoprecipitation assay to validate gene interactions. Rescue experiments were conducted to analyze the role of TRIM16 in CRC radioresistance.
Results
LncRNA OTUD6B-AS1 and TRIM16 were poorly expressed (P < 0.01) in CRC tissues and cells and further decreased (P < 0.01) in radioresistant CRC cells. OTUD6B-AS1 overexpression decreased cell survival (P < 0.01), increased γ-H2AX levels (P < 0.01), and elevated ferroptosis and oxidative stress (P < 0.01) after X-ray radiation. Ferroptosis inhibitor attenuated radioresistance (P < 0.01) caused by lncRNA OTUD6B-AS1 overexpression. LncRNA OTUD6B-AS1 stabilized TRIM16 mRNA via binding to HuR. TRIM16 knockdown reduced ferroptosis and increased radioresistance (P < 0.05).
Conclusion
OTUD6B-AS1 overexpression stabilized TRIM16 via binding to HuR and increased GPX4-mediated ferroptosis, thus attenuating CRC radioresistance. Our study provided a new rationale for the treatment of CRC.
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Data availability
The data that support the findings of this study are available from the corresponding author upon reasonable request.
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Funding
This work was supported by the Special Fund of Foshan Summit Plan (No. 2019C020) and the Construction Funds for High-level Medical Key Specialty Projects in Foshan during the 14th Five Year Plan Period (No. 0660215).
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Conceptualization: ZZ, BY, YL; Formal analysis and investigation: ZZ, WL; Writing—original draft preparation: ZZ, WL, JD; Writing—review and editing: ZZ, WJ. All authors read and approved the final manuscript.
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This study conformed to the Declaration of Helsinki and got the approval of the Ethics Committee of **ling Hospital, the First School of Clinical Medicine, Southern Medical University.
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Zhang, Z., Ye, B., Lin, Y. et al. LncRNA OTUD6B-AS1 overexpression promoted GPX4-mediated ferroptosis to suppress radioresistance in colorectal cancer. Clin Transl Oncol 25, 3217–3229 (2023). https://doi.org/10.1007/s12094-023-03193-7
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DOI: https://doi.org/10.1007/s12094-023-03193-7