Objective  To investigate the mechanism of microRNA-214-3p (miR-214-3p) targeting CHUK to regulate the nuclear factor (NF)-κB pathway in modulating the chemosensitivity of lymphoma 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.