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Circ_0003356 suppresses gastric cancer growth through targeting the miR-668-3p/SOCS3 axis
Wei-Dong Li1, Hai-Tao Wang1, Yue-Ming Huang1
1Department of Gastrointestinal Surgery, Zhongshan City People's Hospital, Zhongshan 528403, Guangdong Province, China.
Background:
Circular RNAs (circRNAs) have attracted extensive attention as therapeutic targets in gastric cancer (GC). Circ_0003356 is known to be downregulated in GC tissues, but its cellular function and mechanisms remain undefined.
Aim:
To investigate the role of circ_0003356 in GC at the molecular and cellular level.
Methods:
Circ_0003356, miR-668-3p, and SOCS3 expression were assessed via quantitative real time-polymerase chain reaction (qRT-PCR). Wound healing, EdU, CCK-8, flow cytometry and transwell assays were used to analyze the migration, proliferation, viability, apoptosis and invasion of GC cells. The subcellular localization of circ_0003356 was monitored using fluorescence in situ hybridization. The interaction of circ_0003356 with miR-668-3p was confirmed using RIP-qRT-PCR, RNA pull-down, and dual luciferase reporter assays. We observed protein levels of genes via western blot. We injected AGS cells into the upper back of mice and performed immunohistochemistry staining for examining E-cadherin, N-cadherin, Ki67, and SOCS3 expressions. TUNEL staining was performed for the assessment of apoptosis in mouse tumor tissues.
Results:
Circ_0003356 and SOCS3 expression was downregulated in GC cells, whilst miR-668-3p was upregulated. Exogenous circ_0003356 expression and miR-668-3p silencing suppressed the migration, viability, proliferation, epithelial to mesenchy-mal transition (EMT) and invasion of GC cells and enhanced apoptosis. Circ_0003356 overexpression impaired tumor growth in xenograft mice. Targeting of miR-668-3p by circ_0003356 was confirmed through binding assays and SOCS3 was identified as a downstream target of miR-668-3p. The impacts of circ_0003356 on cell proliferation, apoptosis, migration, invasion and EMT were reversed by miR-668-3p up-regulation or SOCS3 down-regulation in GC cells.
Conclusion:
Circ_0003356 impaired GC development through its interaction with the miR-668-3p/SOCS3 axis.
Insights
Circular RNA circ_0003356 inhibits gastric cancer (GC) progression by targeting miR-668-3p and regulating SOCS3. This finding offers a potential therapeutic strategy for GC treatment.
Area of Science:
- Molecular Biology
- Cancer Research
- Genetics
Background:
- Circular RNAs (circRNAs) are emerging as critical regulators in various cancers, including gastric cancer (GC).
- Circ_0003356, a specific circRNA, has been observed to be downregulated in GC tissues, yet its functional role remains unclear.
- Understanding the molecular mechanisms of circ_0003356 is crucial for developing novel therapeutic strategies against GC.
Purpose of the Study:
- To elucidate the molecular and cellular functions of circ_0003356 in gastric cancer.
- To investigate the regulatory pathway involving circ_0003356, miR-668-3p, and SOCS3 in GC progression.
- To assess the therapeutic potential of circ_0003356 in preclinical models of GC.
Main Methods:
- Quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting were used to measure gene and protein expression levels.
- Cellular assays including wound healing, EdU, CCK-8, flow cytometry, and transwell assays were employed to evaluate GC cell behavior.
- Fluorescence in situ hybridization, RIP-qRT-PCR, RNA pull-down, and dual luciferase reporter assays were utilized to confirm molecular interactions.
- In vivo studies involved xenograft mouse models to assess tumor growth and apoptosis.
Main Results:
- Circ_0003356 and SOCS3 expression were found to be downregulated, while miR-668-3p was upregulated in GC cells.
- Overexpression of circ_0003356 or silencing of miR-668-3p significantly inhibited GC cell migration, proliferation, viability, invasion, and epithelial-mesenchymal transition (EMT), while promoting apoptosis.
- Circ_0003356 directly targeted miR-668-3p, and miR-668-3p targeted SOCS3, establishing a regulatory axis.
- In vivo experiments demonstrated that circ_0003356 overexpression suppressed tumor growth in xenograft mice.
Conclusions:
- Circ_0003356 plays a tumor-suppressive role in gastric cancer development.
- The inhibitory effect of circ_0003356 on GC is mediated through the miR-668-3p/SOCS3 signaling pathway.
- Circ_0003356 represents a promising therapeutic target for gastric cancer treatment.
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