386P - FOLR1 stabilized beta-catenin promotes laryngeal carcinoma progression through EGFR signal

Background

Wnt/β-catenin pathway is closely related to the development and progression of laryngeal squamous cell carcinoma (LSCC). Loss or reduction of ubiquitination and degradation of β-catenin in cytoplasm result in its accumulation in the nucleus and over-activation of the Wnt/β-catenin pathway. Folate receptor 1 (FRα) is overexpressed in numerous epithelial malignancies. However, the effect of FRα on Wnt/β-catenin pathway and its role in LSCC remains unclear.

Methods

Real-time RT-PCR, Western blot and immunohistochemistry(IHC) detects the mRNA and protein level of FRα in 30 cases of laryngeal squamous cell carcinoma (LSCC) tissues and adjacent normal tissues; Flag-FRα and Si# FRα plasmids were constructed and transfected to establish high FRα expression and low FRα expression cells; EdU assay, colony formation, scratch assay and transwell assay were performed to detect the of proliferation, colony formation, migration and invasion ability of cells; SiRNA#β-catenin was used to block Wnt/β-catenin signal and detected the ability of proliferation, colony formation, migration and invasion; confocal laser detects the subcellular distribution of β-catenin; SgRNA#EGFR was used to silence EGFR signal ; co-immunoprecipitation detects the ubiquitination level of β-catenin.

Results

Q-RT-PCR, WB and IHC showed that the mRNA and protein level of FRα in laryngeal carcinoma tissues was significantly higher than adjacent tissues, and the difference was statistically significant (P<0.01). Compared with FRα low expression (TU177/Flag and TU212/Si-FRα) cells, the proliferation, colony formation, migration and invasion abilities of FRα high expression (TU177/Flag-FRα and TU212/Si-NC) cells were enhanced; SiRNA#β-catenin inhibited FRα -induced cell proliferation, colony formation and migration. Confocal laser results showed that the nuclear accumulation of β-catenin was increased in FRα high expression cells. SgRNA#EGFR inhibited FRα -induced nuclear translocation of β-catenin. Co-immunoprecipitation results displayed that the ubiquitination of β-catenin was decreased in FRα high expression cells.

Conclusions

FOLR1 stabilized β-catenin promotes laryngeal carcinoma progression through EGFR signa.

Clinical trial identification

Editorial acknowledgement

Legal entity responsible for the study

H.Tuo.

Funding

Has not received any funding.

Disclosure

All authors have declared no conflicts of interest.