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Updated: Mar 19, 2026

An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells
Published on: May 21, 2019
miR-20b downregulates polymerases κ and θ in XP-V tumor cells
Jia Guo1, Zheng Jiang2, Xiangru Li1
1Department of Endodontics, Oral Medical Center, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, P.R. China.
Abstract:
XP-V is a subtype of Xeroderma pigmentosum diseases with typical pigmentation and cancers in sun-exposed regions. The present study investigated the role of microRNA-20b (miR-20b) in the imbalance of polymerase expression levels in XP-V tumor cells. Following software prediction results, certain miRNAs were chosen as candidate regulators for the observed imbalance in polymerases in XP-V tumor cells. Reverse transcription-quantitative polymerase chain reaction and western blot were used to test candidate miRNAs for their ability to reduce the expression of these polymerases. A luciferase reporter assay was used to further verify the western blot results. Polymerases κ and θ were expressed at lower levels in XP-V tumor cells compared to normal control cells. A positive correlation was demonstrated between miR-20b and polymerases κ and θ. It was also demonstrated that a proportion of miRNAs had no effect on polymerases κ and θ, despite the software predicting that these miRNAs would target these two polymerases. Therefore, miR-20b may be responsible for the low expression levels of polymerase κ and θ in XP-V tumor cells, which accelerated mismatch in DNA replication repairing.
Insights
MicroRNA-20b (miR-20b) may cause low expression of polymerases κ and θ in Xeroderma pigmentosum variant (XP-V) tumor cells, accelerating DNA replication errors.
Area of Science:
- Molecular Biology
- Genetics
- Oncology
Background:
- Xeroderma pigmentosum variant (XP-V) is characterized by skin cancers.
- Imbalances in polymerase expression are observed in XP-V tumor cells.
Purpose of the Study:
- Investigate the role of microRNA-20b (miR-20b) in polymerase expression dysregulation in XP-V tumors.
- Identify specific microRNAs targeting polymerases κ and θ.
Main Methods:
- Bioinformatic prediction of microRNA regulators.
- Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and Western blot to assess expression levels.
- Luciferase reporter assay for validation.
Main Results:
- Polymerases κ and θ were significantly downregulated in XP-V tumor cells.
- miR-20b showed a positive correlation with polymerases κ and θ expression.
- Some predicted microRNAs did not affect polymerase expression.
Conclusions:
- miR-20b is implicated in the reduced expression of polymerases κ and θ in XP-V tumors.
- This downregulation may contribute to impaired DNA replication repair and cancer development in XP-V.
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