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Updated: Dec 24, 2025

The Soft Agar Colony Formation Assay
Published on: October 27, 2014
Cell proliferation is induced in renal cell carcinoma through miR-92a-3p upregulation by targeting FBXW7
Rong Zeng1, Jing Huang1, Yujie Sun1
1Teaching Experimental Center, School of Laboratory Medicine, Hubei University of Chinese Medicine, Wuhan, Hubei 430065, P.R. China.
Abstract:
Renal cell carcinoma (RCC) is the most common type of kidney cancer whose incidence has gradually increased worldwide. MicroRNAs (miRNAs) represent a type of short endogenous non-coding RNA containing approximately 22 nucleotides, which are capable of regulating mRNAs at the post-transcriptional level in human cells. miRNAs have been demonstrated to mediate gene expression by influencing important regulatory genes. Accumulating evidence indicates that certain miRNAs are involved in RCC development. The present study investigated the underlying mechanism and functional role of miR-92a-3p in RCC cells using reverse transcription-quantitative polymerase chain reaction, western blotting, 3' UTR luciferase assay, cell proliferation assay and soft agar assay. The results demonstrated that miR-92a-3p expression level is significantly upregulated in RCC tissues and cell lines; however, F-box and WD repeat domain containing 7 (FBXW7) expression level was significantly downregulated in RCC tissues and cell lines. Subsequently, whether FBXW7 could be considered as a direct target of miR-92a-3p in RCC cells was investigated. The results demonstrated that miR-92a-3p overexpression significantly promoted RCC cell proliferation and colony formation. Conversely, miR-92a-3p downregulation significantly inhibited RCC cell proliferation and colony formation. In addition, FBXW7 knockdown significantly enhanced RCC cell proliferation and colony formation. Conversely, FBXW7 overexpression significantly inhibited RCC cell proliferation and colony formation. Collectively, these results demonstrated that miR-92a-3p/FBXW7 pathway may represent a novel strategy and therapeutic target for RCC.
Insights
MicroRNAs (miRNAs) like miR-92a-3p are upregulated in kidney cancer, promoting tumor growth by downregulating FBXW7. Targeting this miR-92a-3p/FBXW7 pathway offers a potential new strategy for renal cell carcinoma (RCC) treatment.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Renal cell carcinoma (RCC) incidence is rising globally.
- MicroRNAs (miRNAs) regulate gene expression and are implicated in cancer development.
- Specific miRNAs are increasingly recognized for their roles in RCC pathogenesis.
Purpose of the Study:
- To investigate the mechanism and function of miR-92a-3p in renal cell carcinoma (RCC).
- To explore the relationship between miR-92a-3p and F-box and WD repeat domain containing 7 (FBXW7) in RCC.
- To assess the therapeutic potential of targeting the miR-92a-3p/FBXW7 pathway in RCC.
Main Methods:
- Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) to measure miRNA and gene expression.
- Western blotting to assess protein levels.
- 3' UTR luciferase assay to confirm direct targeting.
- Cell proliferation and soft agar assays to evaluate cellular growth and transformation.
Main Results:
- miR-92a-3p expression was significantly upregulated in RCC tissues and cell lines.
- FBXW7 expression was significantly downregulated in RCC tissues and cell lines.
- miR-92a-3p overexpression promoted RCC cell proliferation and colony formation, while FBXW7 knockdown enhanced these processes.
- FBXW7 overexpression inhibited RCC cell proliferation and colony formation, suggesting it is a direct target of miR-92a-3p.
Conclusions:
- The miR-92a-3p/FBXW7 pathway plays a crucial role in the progression of renal cell carcinoma.
- miR-92a-3p acts as an oncogenic miRNA by targeting and downregulating FBXW7.
- This pathway represents a potential novel therapeutic target for RCC treatment.
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