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circGAPVD1 inhibits the progression of gastric cancer through miR-4424/STK4 axis and encoding GAPVD1-137aa protein
Linqi Zhu1, Zhendong Yao2, Yun Liu3
1Department of Clinical Laboratory, The Third Affiliated Hospital of Soochow University, Changzhou, Jiangsu, China; The Fourth Affiliated Hospital of Jiangsu University, Zhenjiang, Jiangsu, China; The Affiliated Taizhou Second People's Hospital of Yangzhou University, Taizhou, Jiangsu, China.
Background:
Gastric cancer (GC) is a malignant gastrointestinal tumor that originates from the epithelium of the gastric mucosa, and circular RNA (circRNA) plays an important role in its progression.
Methods:
The localization of circGAPVD1 and miR-4424 was determined using cytoplasmic RNA isolation and fluorescence in situ hybridization (FISH) assay. The expression levels of circGAPVD1, miR-4424 and mRNA transcripts were determined by quantitative real-time polymerase chain reaction (qPCR). The biological functions of circGAPVD1, miR-4424, serine/threonine kinase 4 (STK4) and GAPVD1-137aa in GC progression evaluated using Transwell, cell invasion, wound scratch assay, cell counting kit 8 (CCK-8), and flow cytometry assays. Protein expression levels were determined by Western blot analysis and immunofluorescence assay. The sponging interaction between circGAPVD1 and miR-4424 was validated using a dual-luciferase reporter assay. The effect of circGAPVD1 on tumor formation ability in vivo was evaluated in BALB/c nude mice with subcutaneous tumors, and the protein-coding potential was verified by induction and purification of proteins.
Results:
circGAPVD1 and miR-4424 were localized in the nucleus and cytoplasm of GC cells. The expression level of circGAPVD1 in GC tissues was lower than that in paracancerous tissues, and was found to be associated with tumor tissue size and tumor node metastasis (TNM) stage. Survival curve analysis revealed that patients with high circGAPVD1 expression level had better prognosis. circGAPVD1 can adsorb miR-4424 through a sponging mechanism to regulate STK4 expression and suppressing GC cell progression. It was also determined that circGAPVD1 has the ability to encode amino acids and encodes the GAPVD1-137aa protein.
Conclusion:
circGAPVD1 inhibits GC progression via the miR-4424/STK4 axis and encodes the GAPVD1-137aa protein, thereby inhibiting GC progression. This represents the comprehensive study linking circGAPVD1 to GC progression, patient survival outcomes, and therapeutic potential.
Insights
Circular RNA circGAPVD1 inhibits gastric cancer (GC) progression by sponging miR-4424 and regulating STK4. circGAPVD1 also encodes a protein, offering potential therapeutic strategies for GC.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Gastric cancer (GC) is a prevalent malignancy originating from gastric mucosa epithelium.
- Circular RNAs (circRNAs) are increasingly recognized for their roles in cancer progression.
Purpose of the Study:
- To investigate the role of circGAPVD1 in gastric cancer (GC) progression.
- To elucidate the molecular mechanism of circGAPVD1, including its interaction with miR-4424 and STK4.
- To assess the potential of circGAPVD1 as a therapeutic target and its protein-coding ability.
Main Methods:
- Localization of circGAPVD1 and miR-4424 via FISH and RNA isolation.
- Quantification of gene and miRNA expression using qPCR.
- Functional assays including Transwell, invasion, wound scratch, CCK-8, and flow cytometry.
- Western blot and immunofluorescence for protein analysis.
- Dual-luciferase reporter assay for interaction validation.
- In vivo tumor formation assay in nude mice.
Main Results:
- circGAPVD1 and miR-4424 are found in the nucleus and cytoplasm of GC cells.
- Lower circGAPVD1 expression correlates with advanced TNM stage and larger tumor size in GC.
- High circGAPVD1 expression is associated with improved patient prognosis.
- circGAPVD1 acts as a sponge for miR-4424, regulating STK4 expression and inhibiting GC cell progression.
- circGAPVD1 demonstrates protein-coding potential, producing the GAPVD1-137aa protein.
Conclusions:
- circGAPVD1 inhibits GC progression through the miR-4424/STK4 pathway.
- circGAPVD1 encodes the GAPVD1-137aa protein, further contributing to GC inhibition.
- This study highlights circGAPVD1 as a potential biomarker and therapeutic target for gastric cancer.
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