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Updated: Feb 6, 2026
Analysis of Combinatorial miRNA Treatments to Regulate Cell Cycle and Angiogenesis
Published on: March 30, 2019
Upregulated miR-1258 regulates cell cycle and inhibits cell proliferation by directly targeting E2F8 in CRC
Zhiyuan Zhang1, Jie Li1, Yuanjian Huang1
1Department of General Surgery, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, China.
Objectives:
MicroRNAs (miRNAs) as small noncoding RNA molecules function by regulating their target genes negatively. MiR-1258 was widely researched in multicancers, but its role remains unclear in colorectal cancer (CRC).
Methods:
The expression of miR-1258 and its specific target gene were detected in human CRC specimens and cell lines by miRNA RT-PCR, qRT-PCR and Western blot. The effects of miR-1258 on CRC proliferation were evaluated using CCK-8 assays, EdU incorporation, colony formation assays and cell-cycle assays; in vitro and the in vivo effects were investigated using a mouse tumorigenicity model. Luciferase reporter and RIP assays were employed to identify interactions between miR-1258 and its specific target gene.
Results:
MiR-1258 was downregulated in CRC tissues and CRC cell lines, and upregulated miR-1258 was proved to inhibit proliferation and arrest cell cycle at G0/G1 in vitro and vivo. Luciferase reporter, RIP and western blot assays revealed E2F8 to be a direct target of miR-1258. The effects of miR-1258 in proliferation and cell cycle regulation can be abolished by E2F8 through rescue experiments. By directly targeting E2F8, miR-1258 influenced the expression of several cell-cycle factors, including cyclin D1 (CCND1) and cyclin dependent kinase inhibitor 1A (p21).
Conclusion:
MiR-1258 may function as a suppressive factor by negatively controlling E2F8, thus, highlighting the potential role of miR-1258 as a therapeutic target for human CRC.
Insights
MicroRNA-1258 (miR-1258) is downregulated in colorectal cancer (CRC). Restoring miR-1258 inhibits CRC cell proliferation and tumor growth by targeting E2F8.
Area of Science:
- Molecular Biology
- Oncology
- Genetics
Background:
- MicroRNAs (miRNAs) are small noncoding RNAs that regulate gene expression.
- The role of miR-1258 in colorectal cancer (CRC) is not well understood.
- Previous studies have explored miR-1258 in various cancers.
Purpose of the Study:
- To investigate the function of miR-1258 in colorectal cancer.
- To identify the target genes of miR-1258 in CRC.
- To evaluate the therapeutic potential of miR-1258 in CRC.
Main Methods:
- Quantitative real-time PCR (qRT-PCR) and Western blot to assess expression levels.
- Cell proliferation assays (CCK-8, EdU, colony formation) and cell cycle analysis.
- In vivo mouse tumorigenicity model and luciferase reporter assays.
- RNA immunoprecipitation (RIP) assays to confirm target interaction.
Main Results:
- MiR-1258 expression was significantly downregulated in CRC tissues and cell lines.
- Overexpression of miR-1258 inhibited CRC cell proliferation and induced G0/G1 cell cycle arrest in vitro and in vivo.
- E2F8 was identified as a direct target of miR-1258.
- MiR-1258 suppressed CRC progression by negatively regulating E2F8 and its downstream cell cycle factors (CCND1, p21).
Conclusions:
- MiR-1258 acts as a tumor suppressor in colorectal cancer.
- The miR-1258/E2F8 axis plays a critical role in CRC development.
- MiR-1258 represents a potential therapeutic target for colorectal cancer treatment.
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