CircHIPK3′s dual role in promoting angiogenesis and inhibiting apoptosis through FASN mRNA stabilization in gallbladder cancer - Research Study

Sep 24, 2026

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Objective: To investigate the mechanism by which circHIPK3 enhances FASN stability by inhibiting the ubiquitinated degradation of ALYREF on gallbladder cancer (GBC). Methods: RT-qPCR assessed circHIPK3 in GBC tissues and its clinicopathological associations; functional assays included CCK-8, colony formation, flow cytometry, tube formation, and Transwell, with nude mice implantation. Results: circHIPK3 is upregulated, linked to TNM stage and lymph node metastasis, and drives tumor progression via ALYREF–FASN. Conclusion: circHIPK3 stabilizes FASN mRNA by suppressing ALYREF ubiquitination-mediated degradation, promoting angiogenesis and inhibiting apoptosis, identifying biomarker and therapeutic targets.

LanguageEnglish

CategoryHealthcare

FormatPDF

Pages12

Reading Time30 min

AccessFree to download

Introduction

Background on gallbladder cancer (GBC)

Role and characteristics of circRNAs

circHIPK3 and rationale for investigating ALYREF/FASN axis

Article information

Article details and abstract summary

Clinics 80 (2025) 100697 Contents lists available at ScienceDirect Clinics journal [homepage:](homepage: www.elsevier.com/locate/clinsp)[ www.elsevier.com/locate/clinsp](homepage: www.elsevier.com/locate/clinsp) | Original articles
CircHIPK3′s dual role in promoting angiogenesis and inhibiting apoptosis through FASN mRNA stabilization in gallbladder cancer | | | | | --- | --- | --- | --- | | ZhenWei Lia,b , HaiJiu Wang a , Cheng Wang a , ZhiXin Wang a , Zhi Xie a,b,* , JinMing Liu a
a Department of Hepatopancreatobiliary Surgery, Qinghai University Affiliated Hospital, Xining City, Qinghai Province, China b Qinghai Provincial Key Laboratory of Hydatid Disease Research, Xining City, Qinghai Province, China | | | | | A R T I C L E I N F O | | A B S T R A C T | | | Keywords: CircHIPK3
ALYREF
FASN Ubiquitination Gallbladder cancer | | Objective: To investigate the mechanism by which circHIPK3 enhances FASN stability by inhibiting the ubiquitinated degradation of ALYREF on Gallbladder Cancer (GBC).
Methods: RT-qPCR was performed to detect circHIPK3 in GBC tissues and to analyze the relationship between circHIPK3 and clinicopathological characteristics of GBC patients. CCK-8 and colony formation assays were performed to study the proliferation of GBC cells, flow cytometry to detect apoptosis, tube formation assay to testangiogenesis, and Transwell assay to study migration and invasion. A nude mice implantation assay was conducted. CircHIPK3, FASN and SKP2 were assessed using RT-qPCR and western blot. Ubiquitination assay was performed to detect ALYREF ubiquitination, and RNA pull-down and RIP to investigate the binding relationship between circHIPK3, ALYREF and FASN.
Results: CircHIPK3 was upregulated in GBC tissues, with elevated expression correlating with TNM stages and lymph node metastasis. CircHIPK3 knockdown suppressed GBC progression by inhibiting proliferation, angiogenesis, migratory and invasive capacity while promoting apoptosis in vitro, paralleled by reduced tumor growth in vivo. CircHIPK3 stabilized ALYREF via inhibiting ubiquitination-mediated degradation. ALYREF knockdown phenocopied the tumor-suppressive effects of circHIPK3 depletion, whereas ALYREF overexpression rescuedcircHIPK3 knockdown-induced suppression of malignant phenotypes. CircHIPK3 stabilizes FASN mRNA through ALYREF. Overexpressing FASN counteracted the actions of circHIPK3 downregulation on cellular angiogenesis and apoptosis.
Conclusion: CircHIPK3 enhances FASN stability by inhibiting ALYREF ubiquitination degradation, promoting angiogenesis, and inhibiting apoptosis in GBC. This study positions circHIPK3 as a promising prognostic biomarker and therapeutic target in GBC, with ALYREF and FASN emerging as critical mechanistic nodes within this oncogenic pathway. | | Introduction Gallbladder Cancer (GBC) ranks among the most prevalent malignancies ofthe biliary system, yet constitutes a comparatively uncommon malignancy relative to other systemic tumors.1 GBC demonstrates significant geographic and ethnic variability in global incidence patterns, with a pronounced female predominance. Despite its relatively low overall prevalence, the disease is characterized by clinically insidious presentation, aggressive tumor biology, and poor prognosis.2,3 Radical resection may be the most effective strategy for GBC, but it is only applicable to a small proportion of patients with early diagnosed GBC.1 More than half of the new GBC cases are detected after a laparoscopiccholecystectomy performed for an assumed benign ailment, and the majority of patients lack specific clinical manifestations in the early stages of GBC or present with jaundice, abdominal pain, and other signs indistinguishable from gallstones or cholecystitis. GBC has a strong invasive and metastatic ability, easily metastasizing to organs such asthe liver or adjacent colon, which are extremely rich in blood supply, posing a great challenge to both surgi

FAQs

What is the main objective of the study?

To determine how circHIPK3 enhances FASN stability by blocking ubiquitin-mediated degradation of ALYREF in gallbladder cancer.

How were circHIPK3 and related mechanisms investigated?

RT-qPCR measured circHIPK3 levels in GBC tissues, while functional assays (proliferation, apoptosis, angiogenesis, migration, invasion) and in vivo nude mice implantation assessed biological effects. Ubiquitination, RNA pull-down, and RIP were used to study the circHIPK3–ALYREF–FASN relationships.

What were the key findings regarding circHIPK3 in gallbladder cancer?

CircHIPK3 was upregulated in GBC tissues and its higher expression correlated with advanced TNM stage and lymph node metastasis. circHIPK3 knockdown suppressed malignant phenotypes and promoted apoptosis, mediated through ALYREF stabilization of FASN mRNA.

Sep 24, 2026

Uploaded by Stanley