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Non-canonical RNA-binding protein ANXA11 regulates microRNA resorting into small extracellular vesicles to mediate
Yifan Zhang1, Qiang Huang1, Yujie Shen1
1Department of Otorhinolaryngology, Eye & ENT Hospital, Fudan University, Shanghai, China.
Abstract:
The sensitivity of laryngeal squamous cell carcinoma (LSCC) to chemotherapy shows large heterogeneity. The role of miRNA in small extracellular vesicles (sEV) in chemotherapy resistance is under investigation. However, the regulation and sorting mechanism of sEV miRNAs remains unclear. In this study, small RNA sequencing was used to explore miRNA expression profiles in sEV of LSCC after cisplatin stimulation; RNA pull-down, mass spectrometry, and EMSA were used to clarify the binding of candidate RNA-binding protein (RBP) and candidate miRNA. Immunostaining and microRNA fluorescence in situ hybridization were performed to identify how candidate RBP affects miRNA stability and nuclear/cytoplasmic distribution. In vivo experiments were performed to verify the biological functions and response to cisplatin of candidate RBP. We found that cisplatin stimulation induced increased expression of miR-148a-3p and sEV sorting. ANXA11 binds to miR-148a-3p in a sequence-specific manner. ANXA11 inhibits tumor cell proliferation and drug resistance by binding to and retaining miR-148a-3p. Cisplatin stimulation reduced ANXA11 expression and promoted miR-148a-3p efflux through sEV pathways. ANXA11 overexpression reduced in vivo tumor proliferation and cisplatin-resistance. Taken together, ANXA11 mediates cisplatin resistance through sEV miRNA resorting. Mechanically, ANXA11 binds to miR-148a-3p in a sequence-specific manner to regulate its resorting and thus influences tumor proliferation and chemoresistance.
Insights
Chemotherapy resistance in laryngeal cancer is linked to microRNAs (miRNAs) in extracellular vesicles. This study reveals ANXA11 protein retains miR-148a-3p, impacting chemotherapy response and tumor growth.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Laryngeal squamous cell carcinoma (LSCC) exhibits variable chemotherapy sensitivity.
- The role of microRNAs (miRNAs) within small extracellular vesicles (sEVs) in mediating chemotherapy resistance is an emerging area of research.
- Mechanisms regulating sEV miRNA sorting and function remain largely unelucidated.
Purpose of the Study:
- To investigate the impact of cisplatin treatment on miRNA expression profiles within sEVs derived from LSCC.
- To identify RNA-binding proteins (RBPs) involved in the regulation of specific miRNAs implicated in chemoresistance.
- To elucidate the functional role of identified RBPs and their associated miRNAs in LSCC chemoresistance and tumor progression.
Main Methods:
- Small RNA sequencing to profile sEV miRNAs after cisplatin stimulation.
- RNA pull-down, mass spectrometry, and electrophoretic mobility shift assay (EMSA) to confirm RBP-miRNA interactions.
- Immunostaining and fluorescence in situ hybridization to assess RBP effects on miRNA localization and stability.
- In vivo experiments to validate the biological functions of the candidate RBP.
Main Results:
- Cisplatin stimulation upregulated miR-148a-3p expression and enhanced its sorting into sEVs.
- The RBP ANXA11 was identified to bind specifically to miR-148a-3p.
- ANXA11 binding retains miR-148a-3p, inhibiting tumor cell proliferation and conferring cisplatin resistance.
- Cisplatin treatment decreased ANXA11 levels, promoting miR-148a-3p release via sEVs.
- Overexpression of ANXA11 reduced tumor growth and cisplatin resistance in vivo.
Conclusions:
- ANXA11 mediates cisplatin resistance in LSCC by regulating the sorting of miR-148a-3p into sEVs.
- The sequence-specific binding of ANXA11 to miR-148a-3p is crucial for controlling miRNA resorting and influencing tumor proliferation and chemoresistance.
- Findings highlight a novel mechanism involving ANXA11 and sEV-mediated miRNA transport in LSCC chemoresistance.
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