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Updated: Oct 5, 2025

Modeling Oral-Esophageal Squamous Cell Carcinoma in 3D Organoids
Published on: December 23, 2022
Knockdown of UCHL3 inhibits esophageal squamous cell carcinoma progression by reducing CRY2 methylation
Jijun Xue1, Jinyuan Yi2, Xiaolong Zhu3
1Department of Thoracic Surgery, Gansu Provincial Cancer Hospital, Lanzhou , 730050, Gansu, China.
Abstract:
UCHL3 (Ubiquitin carboxyl-terminal hydrolase L3), a member of deubiquitinating enzymes, has been implicated in various cancers. However, the role of UCHL3 in esophageal squamous cell carcinoma (ESCC) remains unknown. In the current study, we aimed to investigate the role of UCHL3 in ESCC growth and migration, and whether UCHL3 could modulate CRY2 methylation through FOXM1. The expression of UCHL3 and CRY2 in ESCC tissues was assessed using qRT-PCR, western blotting and immunohistochemistry (IHC). Cell viability was determined by CCK-8 and colony formation assays. Hoechst 33342 and flow cytometry were used to detect cell apoptosis. Transwell assay was performed to investigate cell migration and invasion. In vivo animal model was used to assess cell tumorigenesis. Methylation-Specific PCR (MSP) was applied to detect CRY2 methylation in the promoter region. The results showed that UCHL3 expression was elevated in ESCC tissues and cells, while CRY2 expression was decreased. UCHL3 silencing inhibited cell viability, invasion, migration and induced cell apoptosis in vitro, repressed tumor growth in vivo, and increased CRY2 expression and decreased FOXM1 expression. In addition, UCHL3 knockdown decreased CRY2 methylation through downregulating FOXM1, leading to an increase in the expression of CRY2. Moreover, CRY2 silencing abolished UCHL3 deficiency-mediated inhibition in cell growth and migration. In summary, this study reveals that knockdown of UCHL3 inhibits ESCC growth and migration by reducing CRY2 methylation through downregulation of FOXM1 expression.
Insights
Ubiquitin carboxyl-terminal hydrolase L3 (UCHL3) promotes esophageal squamous cell carcinoma (ESCC) growth and migration. Silencing UCHL3 inhibits ESCC by reducing CRY2 methylation via FOXM1 downregulation.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Ubiquitin carboxyl-terminal hydrolase L3 (UCHL3) is implicated in various cancers.
- The specific role of UCHL3 in esophageal squamous cell carcinoma (ESCC) is not well understood.
Purpose of the Study:
- Investigate the role of UCHL3 in ESCC progression.
- Determine if UCHL3 modulates CRY2 methylation through FOXM1.
Main Methods:
- Assessed UCHL3 and CRY2 expression in ESCC tissues and cells (qRT-PCR, Western blotting, IHC).
- Evaluated cell viability, apoptosis, migration, and invasion in vitro.
- Utilized in vivo animal models for tumorigenesis assessment.
- Analyzed CRY2 promoter methylation using Methylation-Specific PCR (MSP).
Main Results:
- UCHL3 expression was elevated in ESCC tissues and cells, correlating with decreased CRY2 expression.
- UCHL3 silencing inhibited ESCC cell viability, migration, invasion, and tumor growth, while inducing apoptosis.
- UCHL3 knockdown decreased CRY2 methylation by downregulating FOXM1, increasing CRY2 expression.
- CRY2 silencing counteracted the inhibitory effects of UCHL3 deficiency on ESCC growth and migration.
Conclusions:
- UCHL3 promotes ESCC growth and migration.
- UCHL3 knockdown inhibits ESCC progression by reducing CRY2 methylation via FOXM1 downregulation.
- UCHL3 represents a potential therapeutic target for ESCC treatment.

