LINC00261 suppresses esophageal squamous cell carcinoma proliferation, invasion, and metastasis by targeting the miR-23a-3p/ZNF292 axis

doi:10.12122/j.issn.1673-4254.2025.10.08

Abstract

Abstract:

Objective To evaluate the regulatory effects of lncRNA LINC00261 on proliferation, invasion, and metastasis of esophageal squamous cell carcinoma (ESCC) cells. Methods The differentially expressed RNAs in ESCC were identified using the GSE149612 dataset from the GEO database. PCR was used to detect LINC00261 expression levels in clinical ESCC and normal esophageal tissue samples and in multiple ESCC cell lines and normal esophageal epithelial cells (HEEC). In ESCC cells, the effects of overexpression of LINC00261 on cell proliferation, invasion, metastasis and apoptosis were analyzed using CCK-8 assay, clone formation assay, Transwell assay and flow cytometry. The potential targets of LINC00261 were predicted using bioinformatics tools including ENCORI and verified using dual-luciferase reporter assay and Western blotting. The effects of LINC00261 overexpression on ESCC were confirmed in a nude mouse model bearing ESCC xenograft. Results Analysis of the GSE149612 dataset revealed significantly lower LINC00261 expression in ESCC tissues and cell lines. In cultured ESCC cells, LINC00261 overexpression markedly suppressed cell proliferation, invasion, and metastasis and promoted cell apoptosis. Dual-luciferase reporter assays confirmed that LINC00261 targets the miR-23a-3p/ZNF292 axis. In the tumor-bearing mouse model, LINC00261 overexpression significantly inhibited ESCC xenograft proliferation and metastasis. Conclusion LINC00261 suppresses ESCC progression by targeting the miR-23a-3p/ZNF292 axis, suggesting a potential therapeutic strategy for ESCC treatment.

Key words: esophageal squamous cell carcinoma, LINC00261, miR-23a-3p, ZNF292

Yuan MI, Xuzhe LI, Zhanpeng WANG, Yanjie LIU, Chuntao SONG, Lantao WANG, Lei WANG. LINC00261 suppresses esophageal squamous cell carcinoma proliferation, invasion, and metastasis by targeting the miR-23a-3p/ZNF292 axis[J]. Journal of Southern Medical University, 2025, 45(10): 2118-2125.

Figures/Tables 5

Fig.1 LINC00261 is lowly expressed in esophageal squamous cell carcinoma (ESCC) tissues and cells. A: Changes of each gene in the GSE149612 dataset. B: Expression of each gene in each tissue in the GSE149612 dataset. C: Expression of LINC00261 in ENCORI. D: Expression changes of LINC00261 in clinical ESCC and normal esophageal tissues. E-H: Expression of LINC00261 in ESCC cells and normal esophageal cells. I: Survival analysis of ESCC patients with high-level or low-level LINC00261 expression. *P<0.05 vs HEEC, ***P<0.001 vs Normal.

Fig.2 Overexpression of LINC00261 inhibits proliferation, invasion and metastasis of ESCC cells and promotes cell apoptosis. A: qRT-PCR for detecting expression changes of LINC00261 after overexpression. B: CCK-8 assay fir assessing changes in cell proliferation ability. C: Clone formation assay for assessing cell proliferation ability. D: Transwell assay for assessing changes in cell invasion and metastasis abilities (×100). E: Detection of changes in cell apoptosis by flow cytometry. **P<0.01, ***P<0.001 vs pcDNA.

Fig.3 LINC00261 targets miR-23a-3p/ZNF292 in ESCC. A: Location of LINC00261 in ESCC cells. B: Correlation analysis of LINC00261 and miR-23a-3p in the ENCORI database. C: Prediction of the targeted genes of miR-23a-3p by ENCORI, DIANA and miRDB databases. D: Correlation analysis of miR-23a-3p and ZNF292 in the ENCORI database. E: Correlation analysis of LINC00261 and ZNF292 in the ENCORI database. F: Bioinformatics websites predict the potential binding sequences of LINC00261 with miR-23a-3p and miR-23a-3p with ZNF292. G: Binding of LINC00261 with miR-23a-3p, and miR-23a-3p and ZNF292 verified by luciferase reporter gene assay. H: Changes in the expression of ZNF292 after overexpression of LINC00261. **P<0.01, ***P<0.001 vs pcDNA.

Fig.4 LINC00261 affects proliferation and metastasis of ESCC xenografts in nude mice. A: Weight changes of subcutaneous transplanted tumors in each group. B: Immunohistochemistry for Ki67, Vimentin and ZNF292 in the tumor tissues in each group. *** P<0.001 vs pcDNA.

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