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Updated: Apr 28, 2026
Analysis of Combinatorial miRNA Treatments to Regulate Cell Cycle and Angiogenesis
Published on: March 30, 2019
[miR-497 suppresses proliferation of human cervical carcinoma HeLa cells by targeting cyclin E1]
Jiming Han1, Manpeng Huo1, Mingtao Mu1
1Medical College, Yan'an University, Yan'an 716000, China.
Objective:
To evaluate the effect of miR-497 on proliferation of human cervical carcinoma HeLa cells and target relationship between miR-497 and cyclin E1 (CCNE1).
Methods:
Pre-miR-497 sequences were synthesized and cloned into pcDNATM6.2-GW to construct recombinant plasmid pcDNATM6.2-GW-pre-miR-497 and identified by real-time quantitative PCR (qRT-PCR). In addition, sequences of the wild-type CCNE1 (WT-CCNE1) and mutant CCNE1 (MT-CCNE1) were respectively cloned into pmirGLO vectors. MTT assay was used to explore the impact of miR-497 on the proliferation of HeLa cells. Furthermore, the target effect of miR-497 on the CCNE1 was identified by dual-luciferase reporter assay system, qRT-PCR and Western blotting.
Results:
The recombinant plasmids pcDNATM6.2-GW-pre-miR-497 and pmirGLO-WT-CCNE1, pmirGLO-MT-CCNE1 were successfully constructed, and the miR-497 expression level in HeLa cells transfected with pre-miR-497 was significantly higher than that in the neg-miR group (P<0.05). MTT assay showed that miR-497 could significantly inhibit the proliferation of HeLa cells (P<0.05). A remarkable reduction of luciferase activities of WT-CCNE1 reporter was observed in HeLa cells with pre-miR-497 transfection (P<0.01), and the mRNA and protein expression levels of CCNE1 were down-regulated in HeLa cells transfected with pre-miR-497 (P<0.05).
Conclusion:
Over-expressed miR-497 in HeLa cells could suppress cell proliferation by targeting CCNE1.
Insights
MicroRNA-497 (miR-497) significantly inhibits human cervical carcinoma HeLa cell proliferation. This microRNA targets cyclin E1 (CCNE1), suppressing its expression and impacting cancer cell growth.
Area of Science:
- Molecular Biology
- Oncology
- Gene Regulation
Context:
- Cervical carcinoma is a significant global health concern.
- Understanding the molecular mechanisms underlying cervical cancer progression is crucial for developing targeted therapies.
- MicroRNAs play critical roles in regulating gene expression and are implicated in various cancers.
Purpose:
- To investigate the role of miR-497 in the proliferation of human cervical carcinoma HeLa cells.
- To determine the direct target relationship between miR-497 and cyclin E1 (CCNE1).
Summary:
- The study successfully constructed recombinant plasmids to overexpress miR-497 in HeLa cells.
- MTT assays demonstrated that elevated miR-497 levels significantly inhibit HeLa cell proliferation.
- Dual-luciferase reporter assays, qRT-PCR, and Western blotting confirmed that miR-497 directly targets CCNE1, downregulating its mRNA and protein expression.
Impact:
- This research elucidates a novel regulatory pathway in cervical cancer involving miR-497 and CCNE1.
- The findings suggest that miR-497 could be a potential therapeutic agent for cervical carcinoma.
- Targeting the miR-497/CCNE1 axis may offer a new strategy for cervical cancer treatment.
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