Mechanism of microRNA-214-3p targeting CHUK in chemosensitivity of ibrutinib by regulating nuclear factor-κB pathway
- VernacularTitle:微小RNA-214-3p靶向CHUK调控核因子-κB通路影响伊布替尼化疗敏感性的机制研究
- Author:
Yiqi FANG
1
;
Qianwen BAI
;
Xiaodu XU
;
Bing WU
;
Weiping WU
;
Jing QIAN
Author Information
- Keywords: microRNA-214-3p; lymphoma; ibrutinib; drug resistance; CHUK protein; half-maximal inhibitory concentration; nuclear factor-κB; cell apoptosis
- From: Journal of Clinical Medicine in Practice 2025;29(11):72-78,84
- CountryChina
- Language:Chinese
- Abstract: Objective To investigate the mechanism of microRNA-214-3p(miR-214-3p)targe-ting CHUK to regulate the nuclear factor(NF)-κB pathway in modulating the chemosensitivity of lym-phoma cells to ibrutinib.Methods The half-maximal inhibitory concentration(IC50)was used to verify the ibrutinib-resistant cell model.The expression levels of miR-214-3p and CHUK mRNA in tissues and cells were detected by quantitative real-time polymerase chain reaction(qRT-PCR).Western blotting(WB)was employed to assess CHUK protein levels.A dual-luciferase reporter assay was performed to confirm the direct targeting relationship between miR-214-3p and CHUK.Cell viability was measured using CCK-8 assay.Flow cytometry was used to evaluate cell apoptosis.The expression of the NF-κB p65 signaling pathway was detected by WB.Results The CHUK mRNA and CHUK protein expression levels were higher in ibrutinib-resistant cells than in control cells(P＜0.05).The qRT-PCR results showed that miR-214-3p was downregulated in ibrutinib-resistant cells.The dual-luciferase reporter assay confirmed that miR-214-3p directly targeted CHUK.Transfection of miR-214-3p mimics and knockdown of CHUK(si-CHUK)reduced the number of colony-forming cells,whereas transfection of miR-214-3p inhibitor increased the number of colony-forming cells(P＜0.05).Transfection of miR-214-3p mimics and knockdown of CHUK(si-CHUK)can promote apoptosis,while transfection of miR-214-3p inhibitor can inhibit apoptosis(P＜0.05).Inhibition of the NF-κB p65 pathway was observed in cells transfected with miR-214-3p mimics(P＜0.05).Conclusion MiR-214-3p may regulate the NF-κB p65 pathway by targeting the expression of CHUK,thereby enhancing the chemosensitivity of ibrutinib to lymphoma.
