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Characterization of Functionally Associated miRNAs in Glioblastoma and their Engineering into Artificial Clusters for Gene Therapy
Published on: October 4, 2019
Knockdown of circ_0008344 contributes to radiosensitization in glioma via miR-433-3p/RNF2 axis
Liang DI1, Xiaofei Zhao, Jing Ding
1General Surgery Department, Beijing You'an Hospital, Capital Medical University, Beijing 100069, China.
Abstract:
Numerous studies have identified that circular RNAs (circRNAs) functioned as important regulators in tumor initiation, carcinogenesis, drug or radiation resistance. This study aims to reveal the function of circ_0008344 on radiosensitivity in glioma. The quantitative real-time polymerase chain reaction (qRT-PCR) was implemented for detecting the circ_0008344 and microRNA-433-3p (miR-433-3p) levels. Cell survival intensity and apoptosis were analyzed through colony formation assay and flow cytometry respectively. The protein levels were examined via Western blot. Dual-luciferase reporter assay was exploited for the analysis of target combination. Xenograft models were established in mice for circ_0008344 research in vivo. Our data showed that circ_0008344 level was signally increased in radioresistant glioma tissues and its down-regulation facilitated the susceptibility of glioma cells to radiation. Additionally, we found that circ_0008344 could interact with miR-433-3p and regulated radiosensitivity of glioma cells via sponging miR-433-3p. Ring finger protein 2 (RNF2) was proved to be a target of miR-433-3p and it was regulated by circ_0008344/miR-433-3p axis. The promotion of circ_0008344 knockdown on radiosensitivity was counteracted by RNF2 overexpression in glioma cells. Further experiment in vivo indicated that circ_0008344 down-regulation inhibited glioma growth and acted on miR-433-3p/RNF2 axis to enhance the radiosensitivity in glioma. These evidences manifested that knockdown of circ_0008344 exerted the radiosensitivity-promoting effect on glioma via the miR-433-3p-mediated RNF2 down-regulation, identifying circ_0008344 as a novel diagnostic biomarker of radioresistance and therapeutic target in glioma radiotherapy.
Insights
Knockdown of circ_0008344 enhances glioma radiosensitivity by sponging miR-433-3p and down-regulating RNF2. This circular RNA is a potential biomarker and therapeutic target for glioma radiotherapy.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Circular RNAs (circRNAs) are key regulators in cancer development and treatment resistance.
- Glioma radiosensitivity is a critical factor in treatment outcomes.
- Understanding the role of specific circRNAs in glioma radiosensitivity is crucial for developing novel therapeutic strategies.
Purpose of the Study:
- To investigate the function of circ_0008344 in regulating radiosensitivity in glioma.
- To elucidate the underlying molecular mechanism involving microRNA-433-3p (miR-433-3p) and Ring finger protein 2 (RNF2).
Main Methods:
- Quantitative real-time polymerase chain reaction (qRT-PCR) for circRNA and miRNA expression analysis.
- Cell viability and apoptosis assays (colony formation, flow cytometry) to assess radiosensitivity.
- Western blot for protein level determination.
- Dual-luciferase reporter assay for target interaction validation.
- In vivo xenograft models in mice to evaluate therapeutic efficacy.
Main Results:
- circ_0008344 expression was significantly elevated in radioresistant glioma tissues.
- Down-regulation of circ_0008344 increased glioma cell sensitivity to radiation.
- circ_0008344 acts as a sponge for miR-433-3p, and RNF2 is a downstream target regulated by this axis.
- Knockdown of circ_0008344 enhanced radiosensitivity by down-regulating RNF2 via miR-433-3p, an effect reversed by RNF2 overexpression.
- In vivo studies confirmed that circ_0008344 knockdown inhibited tumor growth and improved radiosensitivity.
Conclusions:
- circ_0008344 promotes radioresistance in glioma by sponging miR-433-3p and consequently up-regulating RNF2.
- Down-regulation of circ_0008344 enhances glioma radiosensitivity through the miR-433-3p/RNF2 pathway.
- circ_0008344 represents a promising diagnostic biomarker for radioresistance and a potential therapeutic target in glioma radiotherapy.
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