MicroRNA-638 inhibits cell proliferation by targeting phospholipase D1 in human gastric carcinoma
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Updated: Apr 5, 2026
An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells
Published on: May 21, 2019
Jiwei Zhang1, Zehua Bian1, Jialiang Zhou2
1Wuxi Oncology Institute, The Affiliated Hospital of Jiangnan University, Wuxi, 214062 China.
Abstract:
MicroRNAs (miRNAs) are a type of small non-coding RNAs that are often play important roles in carcinogenesis, but the carcinogenic mechanism of miRNAs is still unclear. This study will investigate the function and the mechanism of miR-638 in carcinoma (GC). The expression of miR-638 in GC and the DNA copy number of miR-638 were detected by real-time PCR. The effect of miR-638 on cell proliferation was measured by counting kit-8 assay. Different assays, including bioinformatics algorithms (TargetScan and miRanda), luciferase report assay and Western blotting, were used to identify the target gene of miR-638 in GC. The expression of miR-638 target gene in clinical CRC tissues was also validated by immunohistochemical assay. From this research, we found that miR-638 was downregulated in GC tissues compared with corresponding noncancerous tissues (NCTs), and the DNA copy number of miR-638 was lower in GC than NCTs, which may induce the corresponding downregulation of miR-638 in GC. Ectopic expression of miR-638 inhibited GC cell growth in vitro. Subsequently, we identified that PLD1 is the target gene of miR-638 in GC, and silencing PLD1 expression phenocopied the inhibitory effect of miR-638 on GC cell proliferation. Furthermore, we observed that PLD1 was overexpressed in GC tissues, and high expression of PLD1 in GC predicted poor overall survival. In summary, we revealed that miR-638 functions as a tumor suppressor in GC through inhibiting PLD1.
Insights
MicroRNA 638 (miR-638) acts as a tumor suppressor in gastric cancer (GC). This study found miR-638 inhibits GC cell growth by targeting PLD1, suggesting its potential as a therapeutic target.
Area of Science:
- Molecular Biology
- Oncology
- Genetics
Background:
- MicroRNAs (miRNAs) are crucial in carcinogenesis, but their specific roles and mechanisms in gastric cancer (GC) remain incompletely understood.
- Investigating novel miRNAs like miR-638 is essential for elucidating GC pathogenesis and identifying potential therapeutic targets.
Purpose of the Study:
- To investigate the function and mechanism of microRNA-638 (miR-638) in gastric cancer (GC).
- To identify the direct target gene of miR-638 and its role in GC progression.
Main Methods:
- Real-time PCR and DNA copy number analysis were used to assess miR-638 expression in GC tissues.
- Cell proliferation was evaluated using the CCK-8 assay.
- Bioinformatics, luciferase reporter assays, and Western blotting identified PLD1 as the target gene of miR-638.
- Immunohistochemistry validated PLD1 expression in clinical GC tissues.
Main Results:
- miR-638 was significantly downregulated in GC tissues compared to noncancerous tissues, correlated with decreased DNA copy number.
- Overexpression of miR-638 inhibited GC cell proliferation in vitro.
- PLD1 was identified as a direct target of miR-638, and its silencing mimicked the anti-proliferative effects of miR-638.
- PLD1 was overexpressed in GC tissues and associated with poor patient survival.
Conclusions:
- miR-638 functions as a tumor suppressor in gastric cancer.
- The tumor-suppressive role of miR-638 in GC is mediated through the inhibition of its target gene, PLD1.
- These findings highlight miR-638 and PLD1 as potential biomarkers and therapeutic targets for GC.
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