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Updated: Jan 27, 2026
Development of Compendium for Esophageal Squamous Cell Carcinoma
Published on: April 12, 2024
[MicroRNA-133b suppresses cell proliferation and invasion of esophageal squamous cell carcinoma via downregulating
1Department of Endocrinology, Henan Provincial People's Hospital, Fuwai Central China Cardiovascular Hospital, Zhengzhou 450003, China.
Abstract:
Objective: To investigate the expression of microRNA-133b (miR-133b) in esophageal squamous cell carcinoma (ESCC), and explore its effect and the underlying molecular mechanisms on cell proliferation and invasion. Methods: Real-time quantitative PCR (qPCR) was used to examine miR-133b expression in 63 ESCC tissues and paired adjacent non-cancerous tissues, several ESCC cells (Eca109, EC9706, EC1, TE1, KYSE70) and normal esophageal epithelial cell Het-1A. MiR-133b mimic, inhibitor and negative control (NC) were transfected into TE1 cells. The effect of miR-133b on cell proliferation and invasion were determined by CCK-8 and Transwell assays, respectively. Subsequently, the target gene of miR-133b was predicted by online tools TargetScan and miRDB, which was verified by dual luciferase reporter assays. Finally, Western blot was utilized to detect the effects of miR-133b overexpression on expression of target gene TAGLN2 as well as EMT-related proteins E-cadherin, N-cadherin, Snail, Slug and Vimentin. Results: Relative levels of miR-133b in ESCC tissues (0.295±0.040) were significantly lower than those in adjacent non-cancerous tissues (1.002±0.011, P<0.001). The expression of miR-133b was tightly associated with clinical staging, lymph node metastasis and prognosis. Moreover, relative levels of miR-133b in ESCC cells Eca109, EC9706, EC1, TE1 and KYSE70 (0.679±0.031, 0.391±0.008, 0.236±0.016, 0.031±0.005 and 0.099±0.020) were evidently lower than that in normal esophageal epithelial cell Het-1A (1.005±0.016, all P<0.001). In TE1 cells, miR-133b mimic significantly increased the level of miR-133b to 6.199±0.627, and suppressed cell proliferation and invasion, whereas miR-133b inhibitor obviously decreased its expression to 0.182±0.023, and promoted cell proliferation and invasion. Most notably, the relative luciferase activities of miR-133b-mimic group (0.320±0.018) in TE1 cells transfected with TAGLN-3'UTR-WT were markedly lower than that in NC group (1.010±0.036, P<0.001), whereas those in TAGLN-3'UTR-MUT transfection cells were 1.019±0.056 and 1.008±0.021, respectively, showing no significantly statistical difference (P>0.05). Furthermore, miR-133b overexpression markedly downregulated TAGLN2, N-cadherin, Snail, Slug and Vimentin levels, and increased E-cadherin expression. Conclusion: MiR-133b plays an important role in the proliferation and invasion of ESCC cells by regulating TAGLN2 expression, and it may be a potential therapeutic target for ESCC patients.
Insights
MicroRNA-133b (miR-133b) is significantly downregulated in esophageal squamous cell carcinoma (ESCC) tissues and cells. Restoring miR-133b suppresses ESCC cell proliferation and invasion by targeting TAGLN2.
Area of Science:
- Molecular Biology
- Oncology
- Gene Regulation
Background:
- Esophageal squamous cell carcinoma (ESCC) is a major global health concern with complex molecular underpinnings.
- MicroRNAs (miRNAs) are critical regulators of gene expression implicated in various cancers, including ESCC.
- The specific role and regulatory mechanisms of microRNA-133b (miR-133b) in ESCC progression remain incompletely understood.
Purpose of the Study:
- To investigate the expression levels of miR-133b in ESCC tissues and cell lines.
- To elucidate the functional impact of miR-133b on ESCC cell proliferation and invasion.
- To identify the molecular targets and pathways regulated by miR-133b in ESCC.
Main Methods:
- Quantitative real-time PCR (qPCR) to assess miR-133b expression in ESCC tissues and cell lines.
- Transfection of miR-133b mimics and inhibitors into ESCC cells (TE1) to modulate its levels.
- Cell proliferation (CCK-8) and invasion (Transwell) assays to evaluate functional effects.
- Bioinformatic prediction (TargetScan, miRDB) and dual-luciferase reporter assays to identify miR-133b targets.
- Western blot analysis to examine the expression of target gene TAGLN2 and epithelial-mesenchymal transition (EMT)-related proteins.
Main Results:
- miR-133b expression was significantly downregulated in ESCC tissues and cell lines compared to normal controls.
- Downregulation of miR-133b correlated with advanced clinical staging, lymph node metastasis, and poorer prognosis.
- Overexpression of miR-133b inhibited ESCC cell proliferation and invasion, while inhibition of miR-133b promoted these processes.
- TAGLN2 was identified as a direct target of miR-133b, with miR-133b negatively regulating TAGLN2 expression.
- miR-133b overexpression led to decreased expression of TAGLN2, N-cadherin, Snail, Slug, and Vimentin, and increased E-cadherin expression.
Conclusions:
- miR-133b functions as a tumor suppressor in ESCC by inhibiting cell proliferation and invasion.
- The tumor-suppressive role of miR-133b is mediated, at least in part, through the regulation of TAGLN2 expression and the EMT pathway.
- miR-133b represents a potential therapeutic target for the treatment of esophageal squamous cell carcinoma.
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