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Published on: September 15, 2023
MiR-769-5p functions as an oncogene by down-regulating RYBP expression in gastric cancer
1Department of General Surgery, Yantaishan Hospital, Yantai, China. yt6709936@163.com.
Objective:
The purpose of this study was to detect the relative expression level of micro-ribonucleic acid (miR)-769-5p in gastric cancer (GC) tissues and cells, and to investigate the clinical significance, biological function, and mechanism of miR-769-5p in GC.
Patients And Methods:
The relative expression level in 62 cases of GC tissues and paracancerous tissues was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The correlation between miR-769-5p expression and clinicopathological characteristics of GC patients was analyzed by Chi-square test. Besides, the relative expression level of miR-769-5p in GC cells and the interference efficiency of si-miR-769-5p were detected by qRT-PCR, and the biological function of miR-769-5p was studied by in vitro experiments [Thiazolyl Blue Tetrazolium Bromide (MTT), flow cytometry, 5-ethynyl-2'-deoxyuridine (EdU)]. Next, the effect of miR-769-5p on the tumorigenicity of GC cells in vivo was investigated by nude mouse tumorigenicity assay. Moreover, the downstream target genes of miR-769-5p were predicted by bioinformatics. Finally, qRT-PCR and Western blotting were used to screen the downstream target genes.
Results:
In the 62 cases of GC tissues, the expression of miR-769-5p was upregulated in 48 cases. MiR-769-5p was divided into high-expression group and low-expression group. Chi-square analysis showed that the high expression of miR-769-5p was positively correlated with tumor-node-metastasis (TNM) stage (p=0.005), lymph node metastasis (p=0.010), and infiltration depth (p=0.011) in patients with GC. The results of qRT-PCR indicated that the expression of miR-769-5p was upregulated in GC cells. In vitro experiments (MTT, flow cytometry, EdU) results showed that after interfering in the expression of miR-769-5p, the proliferation ability of GC cells was decreased, and apoptosis was increased. Furthermore, the results of in vivo experiments manifested that the tumorigenic ability of GC cells declined after interference in the expression of miR-769-5p. Finally, the results of qRT-PCR and Western blotting revealed that the expression of RING1 and YY1-binding protein (RYBP) was regulated by miR-769-5p.
Conclusions:
The expression of miR-769-5p is upregulated in GC and positively correlated with TNM stage in GC patients. By regulating the expression of RYBP, the proliferation of GC cells was promoted, and the apoptosis was inhibited.
Insights
MicroRNA-769-5p (miR-769-5p) is upregulated in gastric cancer (GC) and linked to advanced TNM stage. This microRNA promotes GC cell proliferation and inhibits apoptosis by regulating RYBP expression.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Gastric cancer (GC) remains a significant global health challenge.
- MicroRNAs (miRNAs) play crucial roles in cancer development and progression.
- Understanding the specific roles of miRNAs like miR-769-5p in GC is essential for developing targeted therapies.
Purpose of the Study:
- To determine the expression level of miR-769-5p in gastric cancer tissues and cells.
- To investigate the clinical significance, biological function, and underlying mechanism of miR-769-5p in GC.
- To identify downstream target genes regulated by miR-769-5p.
Main Methods:
- Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) for miRNA expression analysis.
- In vitro assays (MTT, flow cytometry, EdU) to assess cell proliferation and apoptosis.
- In vivo nude mouse tumorigenicity assay to evaluate tumor growth.
- Bioinformatics prediction and Western blotting to identify and validate target genes.
Main Results:
- miR-769-5p was significantly upregulated in 48 out of 62 GC tissues.
- High miR-769-5p expression correlated positively with TNM stage, lymph node metastasis, and tumor infiltration depth.
- In vitro and in vivo studies demonstrated that inhibiting miR-769-5p reduced GC cell proliferation and tumorigenicity while increasing apoptosis.
- RING1 and YY1-binding protein (RYBP) was identified as a downstream target regulated by miR-769-5p.
Conclusions:
- miR-769-5p is overexpressed in gastric cancer and associated with advanced disease stages.
- miR-769-5p promotes GC cell proliferation and inhibits apoptosis, partly through the regulation of RYBP.
- These findings highlight miR-769-5p as a potential therapeutic target for gastric cancer.
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