Work

Antimicrobial peptide WK-13-3D promotes apoptosis, autophagy, and ubiquitination in triple-negative breast cancer via binding immunoglobulin protein (BiP)

Year: 2025

Type: article

Abstract: To elucidate the inhibitory mechanism of antimicrobial peptide WK-13-3D on triple-negative breast cancer (TNBC) by targeting the binding immunoglobulin protein (BiP), a key endoplasmic reticulum (ER) chaperone regulating unfolded protein response and tumor survival. TNBC cell lines (MDA-MB-231 and MDA-MB-468) were treated with WK-13-3D to assess proliferation, migration, invasion, and apoptosis. Pull-down assays identified interacting proteins, and Western blotting (WB) analyzed alterations in BiP, PERK, eIF2α, p -eIF2α, Caspase3, Cleaved-Caspase3, Bax, LC3, P62, AKT, p -AKT, mTOR, and p -mTOR. Transmission electron microscopy examined intracellular structures, while qPCR measured BiP mRNA levels. The effects of WK-13-3D on BiP ubiquitination were explored via co-immunoprecipitation (Co-IP). Animal tumor models were used to confirm the inhibitory effects, with BiP and Ki67 (a nuclear proliferation marker indicating actively dividing tumor cells) expression analyzed by immunohistochemistry (IHC). WK-13-3D inhibited TNBC cell proliferation, migration, and invasion, while promoting apoptosis. Pull-down experiments identified 268 interacting proteins, with BiP being the most frequent. Databases (TIMER and TCGA) showed high BiP expression in breast cancer, associated with poor prognosis. WB assays revealed that WK-13-3D activated ER stress-induced apoptosis and autophagy via BiP. Co-IP demonstrated that WK-13-3D mediated BiP ubiquitination at sites 352 and 547 through K6 and K29 chains. IHC analysis further confirmed decreased Ki67 levels in WK-13-3D-treated tumors, reflecting suppressed proliferative activity. Animal experiments confirmed tumor growth inhibition. WK-13-3D promotes apoptosis, autophagy and Ubiquitination in TNBC by modulating BiP. • A pull-down assay revealed that BiP is the most frequently matched protein interacting with WK-13-3D. • WK-13-3D activated endoplasmic reticulum stress-induced apoptosis and autophagy in TNBC cells via the PERK-eIF2α signaling pathway. • Co-immunoprecipitation showed that WK-13-3D mediated BiP ubiquitination at residues 352 and 547 through K6 and K29 ubiquitin chains. • WK-13-3D holds promise for further development as a novel therapeutic strategy for TNBC by modulating BiP-mediated ER stress.

Source: Chemico-Biological Interactions

DOI: 10.1016/j.cbi.2025.111530

Authors: Wenjing Zhang, Fei Ma, Xuhong Su, Mingxing Zhu, Xiuqing Wang

Institution: Ningxia Medical University

Language: English

Topic: Endoplasmic Reticulum Stress and Disease

Subfield: Cell Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: hybrid

Estimated APC paid: $3,540