miR-96-5p antagonizes FOXQ1-driven WNT/β-catenin signaling to inhibit triple-negative breast cancer

Sep 22, 2026

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FOXQ1 overexpression and Wnt/β-catenin pathway hyperactivation drive triple-negative breast cancer (TNBC), but their functional relationship was unclear. Using TCGA data and RT-PCR, western blotting, and Transwell assays, the study assessed FOXQ1 expression, clinical relevance, and oncogenic function. Integrating bioinformatics with CCLE analyses, RT-qPCR, luciferase reporter assays, and pathway inhibition (IWP-2), it defined a FOXQ1-WNT2-β-catenin positive feedback loop. miR-96-5p was screened and validated as a direct FOXQ1 3’UTR-target, suppressing TNBC tumor growth and aggressiveness in vitro and in vivo while improving prognosis.

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miR-96-5p antagonizes FOXQ1-driven WNT/β-catenin signaling to inhibit triple-negative breast cancer

FOXQ1 expression, clinical relevance, and oncogenic role in TNBC

Transcriptional regulation of FOXQ1 on WNT2 and signaling interplay

miR-96-5p identification and functional validation

[www. nature.com/scientificreports](www. nature.com/scientificreports) OPEN miR-96-5p antagonizes FOXQ1-driven WNT/β-catenin signaling to inhibit triple-negative breast cancer Zhuoran Zhang1,2, Chenshan Zhang1, Yuxuan Meng1, Xianyu Zhu1, Jianying Yang3, Bing Ran4, Zhonglin Gan1, Kejiang Wang5, Mao Yang1, Pinjun Lu1, Sakorn Pornprasert6, Tao He1􀀍 & Yan Lin1􀀍 FOXQ1 overexpression and Wnt/β-catenin pathway hyperactivation are implicated inTNBC, but their functional interconnection is poorly defined. The expression pattern, clinical significance, and oncogenic role of FOXQ1 were systematically assessed using data from The Cancer Genome Atlas (TCGA) combined with RT-PCR, western blotting, and Transwell assays. To explore the transcriptional regulation of FOXQ1 on WNT2, we integrated bioinformatic predictions with analyses of the CCLE dataset, RT-qPCR, western blotting, and luciferase reporter assays. The functional interplay within the FOXQ1-WNT2-β-catenin signaling axis was evaluated through immunofluorescence staining, TOP/ FOP flash reporter assays, and pharmacological inhibition using the Wnt pathway inhibitor IWP-2 . miR-96-5p was identified through multi-database screening and experimentally validated by 3’UTR reporter assays, functional experiments, and in vivo xenograft models. FOXQ1 is upregulated inTNBC and correlates with poor patient survival (HR = 1.34, p = 0.041). It promotes epithelial-mesenchymal transition (EMT), cell migration, and invasion inTNBC. Mechanistically, FOXQ1 directly transactivates WNT2, leading to enhanced nuclear translocation and transcriptional activation of β-catenin. Although IWP-2 inhibits β-catenin activation, it does not prevent FOXQ1-induced upregulation of WNT2, indicating that FOXQ1 acts both upstream of and independently within the Wnt signaling pathway. Furthermore, miR-96-5p directly targets the 3’UTR of FOXQ1, is associated with an improved prognosis (HR = 0.82, p = 0.048), and suppressesTNBC tumor growth and aggressiveness in vitro and in vivo. In conclusion, we have identified a FOXQ1-WNT2-β-catenin positive feedback loop as a critical driver ofTNBC pathogenesis and propose miR-96-5p as a promising therapeutic strategy to suppress this signaling pathway. Keywords Triple-negative breast cancer, FOXQ1, miR-96-5p, WNT2, Wnt/β-catenin signaling, Feedback loop Abbreviations TNBC Triple-negative breast cancer EMT Epithelial–mesenchymal transition Wnt Wingless/integrated miR MicroRNA RNA Ribonucleic acid GSK-3β Glycogen synthase kinase 3 beta Dvl Dishevelled 1Institute for Cancer Medicine, School of Basic Medical Sciences, Southwest Medical University, No.1 Section 1, Xiang Lin Road, Longmatan District, Luzhou 646000, Sichuan, China. 2Academic Affairs Office and Higher Education Research Institution, Southwest Medical University, Luzhou 646000, Sichuan, China. 3Department of Physiology, School of Basic Medical Sciences, Southwest Medical University, Luzhou 646000, Sichuan, China. 4Functional Laboratory, School of Basic Medical Sciences, Southwest Medical University, Luzhou 646000, Sichuan, China. 5Department of Laboratory Medicine, Meishan City People’s Hospital, Meishan 620000, Sichuan, China. 6Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, Thailand. 􀀍 email: [hetao198@swmu.edu.cn](hetao198@swmu.edu.cn); [lydemail@swmu.edu.cn](lydemail@swmu.edu.cn) [www. nature.com/scientificreports/](www. nature.com/scientificreports/) mRNA 3’UTR 3’GAPDH PBS GO Messenger-ribonucleic acid Untranslated region Glyceraldehyde-3-phosphate dehydrogenase Phosphate buffer saline Gene ontology Breast cancer is the leading cause of cancer incidence and cancer-related mortality among women, with 2.3 million new cases and 665,000 deaths reported globally in 20221. Triple-negative breast cancer (TNBC), accounting for 15%-20% of all breast cancers2, is distinguished by its highly aggressive nature. Due to

FAQs

What role does FOXQ1 play in TNBC according to the study?

FOXQ1 is upregulated in TNBC and correlates with poor patient survival. It promotes EMT, migration, and invasion, and it directly enhances WNT2 transcription to activate β-catenin signaling.

How is the FOXQ1-WNT2-β-catenin pathway regulated or interconnected?

FOXQ1 transactivates WNT2, increasing nuclear translocation and transcriptional activation of β-catenin. IWP-2 blocks β-catenin activation but does not prevent FOXQ1-driven WNT2 upregulation, indicating FOXQ1 acts upstream and independently within the pathway.

What is the function of miR-96-5p in this TNBC model?

miR-96-5p directly targets the 3’UTR of FOXQ1 and is associated with improved prognosis. It suppresses TNBC tumor growth and aggressiveness in vitro and in vivo.

Sep 22, 2026

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