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An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells
Published on: May 21, 2019
MicroRNA expression profiling and target gene analysis in gastric cancer
Chenguang Xu1,2, Juan Xie3,4, Yanping Liu1
1Department of Gastroenterology, the Second Affiliated Hospital, University of South China, Hengyang.
Abstract:
This study aims to identify differentially expressed microRNAs (miRNAs) in gastric cancer by comparing gastric cancerous tissues with normal tissues, explore the potential roles.The miRNA expression microarray was employed on gastric cancer tissues, and apparently normal para-cancerous tissues from 3 patients undergoing radical surgery were matched. Quantitative RT-PCR was performed on the other 7 patients to validate the findings of the microarray. Furthermore, Gene Ontology (GO) analysis and enrichment analysis of KEGG Pathway were performed for 5 dysregulated candidate miRNAs, including 3 upregulated (miR-31-3p, miR-6736-3p, and miR-147b) and 2 downregulated (miR-3065-5p and miR-3921) miRNAs, in order to determine the role of miRNAs in tumorigenesis and development.Among these miRNAs, 17 miRNAs were found to be upregulated, and 19 miRNAs were found to be downregulated. The dysregulated expression of 5 candidate miRNAs, including miR-31-3p, miR-147b, miR-6736-3p, miR-3065-5p, and miR-3921, were verified by quantitative RT-PCR in the validation set. Among these miRNAs, miR-31-3p, miR-6736-3p, miR-3065-5p, and miR-3921 had 551 target gene intersections. The GO and KEGG Pathway analyses Revealed that miR-31-3p, miR-6736-3p, miR-3065-5p, and miR-3921 may participate in multiple pathophysiological processes, such as foreign substance metabolism and chemical carcinogenesis.The profile of differentially expressed miRNAs was successfully screened, and 4 miRNAs (i.e., miR-31-3p, miR-6736-3p, miR-3065-5p, and miR-3921) appeared to be involved in gastric carcinogenesis. These might serve as promising biomarkers for gastric cancer.
Insights
This study identified specific microRNAs (miRNAs) dysregulated in gastric cancer. Four key miRNAs show potential as diagnostic biomarkers for this disease.
Area of Science:
- Molecular Biology
- Oncology
- Genetics
Background:
- Gastric cancer is a significant global health concern.
- Understanding the molecular mechanisms, including microRNA (miRNA) dysregulation, is crucial for diagnosis and treatment.
- Identifying novel biomarkers can improve early detection and patient outcomes.
Purpose of the Study:
- To identify differentially expressed microRNAs (miRNAs) in gastric cancer tissues compared to normal tissues.
- To explore the potential roles of these dysregulated miRNAs in gastric carcinogenesis.
- To validate candidate miRNAs as potential biomarkers for gastric cancer.
Main Methods:
- MicroRNA expression microarray analysis on paired gastric cancer and normal tissues.
- Quantitative reverse transcription polymerase chain reaction (qRT-PCR) for validation.
- Gene Ontology (GO) and KEGG Pathway analysis for functional insights into candidate miRNAs.
Main Results:
- 17 miRNAs were upregulated, and 19 were downregulated in gastric cancer tissues.
- Expression of five candidate miRNAs (miR-31-3p, miR-147b, miR-6736-3p, miR-3065-5p, miR-3921) was validated by qRT-PCR.
- Four miRNAs (miR-31-3p, miR-6736-3p, miR-3065-5p, miR-3921) showed significant target gene intersections and involvement in processes like chemical carcinogenesis.
Conclusions:
- A distinct profile of differentially expressed miRNAs in gastric cancer was identified.
- Four specific miRNAs (miR-31-3p, miR-6736-3p, miR-3065-5p, and miR-3921) are implicated in gastric carcinogenesis.
- These miRNAs represent promising potential biomarkers for gastric cancer diagnosis.
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