Related Experiment Video
Updated: Dec 31, 2025
An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells
Published on: May 21, 2019
MicroRNA-769-5p suppresses cell growth and migration via targeting NUSAP1 in bladder cancer
Yifan Chen1, Wentao Zhang1, Aimaitiaji Kadier1
1Department of Urology, Shanghai Tenth People's Hospital, Tongji University, Shanghai, China.
Background:
Nucleolar and spindle-associated protein 1 (NUSAP1) has been identified to be strongly implicated in the carcinogenesis of cervical carcinoma, breast cancer, and liver cancer, and shows a high expression level in bladder cancer, indicating that NUSAP1 might be a potent target for cancer treatment. Using bioinformatics methods, we found that NUSAP1 was a putative target of miR-769-5p. Here, we aimed to explore whether miR-769-5p is involved in bladder cancer progression via targeting NUSAP1.
Methods:
MiR-769-5p expression patterns in bladder cancer tissues and cells were detected by RT-PCR. Kaplan-Meier was used to determine the clinical effects of miR-769-5p expression levels on the overall survival of bladder cancer patients. Bioinformatics methods were used to predict the binding sites between miR-769-5p and NUSAP1, which was verified by the luciferase gene reporter assay. CCK-8, flow cytometry, wound healing and transwell chamber experiments were performed to test cell growth, apoptosis, migration and invasion capacities.
Results:
miR-769-5p was lowly expressed in bladder cancer tissues and cells, which was closely associated with poor prognosis. Overexpression of miR-769-5p induced significant repressions in cell growth, migration, and invasion and caused an obvious increase in cell apoptosis, whereas these tendencies were reversed when NUSAP1 was upregulated.
Conclusion:
This study demonstrates that miR-769-5p functions as a tumor suppressor in bladder cancer via targeting NUSAP1.
Insights
MicroRNA-769-5p acts as a tumor suppressor in bladder cancer by targeting NUSAP1. Low miR-769-5p expression correlates with poor prognosis, while its overexpression inhibits cancer cell growth and invasion.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Nucleolar and spindle-associated protein 1 (NUSAP1) is implicated in various cancers and highly expressed in bladder cancer.
- NUSAP1 is a potential therapeutic target for bladder cancer treatment.
- Bioinformatics analysis identified miR-769-5p as a potential regulator of NUSAP1.
Purpose of the Study:
- To investigate the role of miR-769-5p in bladder cancer progression.
- To determine if miR-769-5p targets NUSAP1 in bladder cancer cells.
Main Methods:
- RT-PCR to detect miR-769-5p expression in bladder cancer tissues and cells.
- Kaplan-Meier analysis for overall survival correlation.
- Luciferase reporter assay to confirm miR-769-5p and NUSAP1 interaction.
- Cell proliferation, apoptosis, migration, and invasion assays (CCK-8, flow cytometry, wound healing, Transwell).
Main Results:
- miR-769-5p was significantly downregulated in bladder cancer tissues and cells, associated with poor prognosis.
- Overexpression of miR-769-5p suppressed bladder cancer cell growth, migration, and invasion, and induced apoptosis.
- Upregulation of NUSAP1 reversed the tumor-suppressive effects of miR-769-5p.
Conclusions:
- miR-769-5p functions as a tumor suppressor in bladder cancer.
- The tumor-suppressive role of miR-769-5p is mediated through the targeting of NUSAP1.
Related Concept Videos
01:22MicroRNAs
01:22MicroRNAs
02:23Abnormal Proliferation