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Published on: October 4, 2019
Knockdown lncRNA CRNDE enhances temozolomide chemosensitivity by regulating autophagy in glioblastoma
Zijin Zhao1,2,3, Miaomiao Liu4, Wenyong Long1,2,3
1Department of Neurosurgery, Xiangya Hospital, Central South University, 87 Xiangya Road, Changsha, 410008, Hunan, China.
Background:
The regulatory roles of long non-coding RNA (lncRNA) CRNDE in temozolomide (TMZ) chemoresistance to glioblastoma multiforme (GBM) are still poorly understood. Therefore, the function, characteristics, and possible mechanism of CRNDE in TMZ-induced chemoresistance to GBM were explored.
Methods:
Firstly, the expression level of CRNDE in 58 cases of glioma tissue specimens and 30 cases of normal brain tissues were tested by qRT-PCR. Meanwhile, the correlation between CRNDE expression level, the clinicopathological characteristics, and survival time of patients with glioma were analyzed. Then, the CRNDE expression in various glioma cell lines was detected, and CRNDE knockdown cell models were constructed. Subsequently, to explore the effect of CRNDE on chemosensitivity to TMZ, cell viability was detected by the CCK-8 assay and IC50 values, and cell proliferation was detected by cell clone assay and EdU assay, as well as cell survival was detected by apoptosis with flow cytometry under TMZ treatment. Further, the expression of drug-resistance protein ABCG2, autophagy related proteins, and PI3K/Akt/mTOR pathway were measured by western blot or qRT-PCR in TMZ-treated glioma cells. Finally, the mouse tumor xenograft model was established and the tumor volume and weight were measured, and ABCG2 expression was conducted by immunohistochemistry assay.
Results:
The integrated results demonstrated lncRNA CRNDE was a poor prognosis factor for GBM patient, which was upregulated in patients who were resistant to TMZ, and closely associated with chemotherapeutic response status to TMZ treatment. Further, functional assays revealed that knockdown of CRNDE could notably reduce glioma cell viability and proliferation, and elevate cell apoptosis to enhance the chemosensitivity to TMZ in vitro and in vivo. Mechanistically, the depression of CRNDE could diminish the expression of LC3 II/I, Beclin1 and Atg5 and increase the p62 expression level to inhibit autophagy due to the activation of PI3K/Akt/mTOR pathway as well as highly correlated with ABCG2 expression.
Conclusions:
Overall, the study provided that lncRNA CRNDE is a reliable clinical predictor of outcome and prognosis and a potential biomarker for predicting TMZ treatment response in GBM by modulating the autophagy through PI3K/Akt/mTOR pathway and ABCG2 expression which may be a novel therapeutic target for regulating TMZ sensitivity to GBM.
Insights
Long non-coding RNA CRNDE promotes temozolomide resistance in glioblastoma by inhibiting autophagy and upregulating ABCG2. Knockdown of CRNDE enhances chemo-sensitivity, suggesting CRNDE as a therapeutic target.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Glioblastoma multiforme (GBM) chemoresistance to temozolomide (TMZ) is a significant clinical challenge.
- The role of long non-coding RNA (lncRNA) CRNDE in TMZ resistance in GBM remains poorly understood.
- Investigating CRNDE's function and mechanism in TMZ resistance is crucial for improving GBM treatment.
Purpose of the Study:
- To explore the function, characteristics, and mechanism of lncRNA CRNDE in TMZ-induced chemoresistance in GBM.
- To evaluate CRNDE as a potential clinical predictor and biomarker for TMZ treatment response in GBM.
- To investigate CRNDE's role in modulating autophagy and ABCG2 expression in GBM.
Main Methods:
- Quantitative real-time PCR (qRT-PCR) to assess CRNDE expression in glioma tissues and cell lines.
- CRNDE knockdown models were established to study its functional impact on chemosensitivity.
- Cell viability, proliferation, and apoptosis assays were performed under TMZ treatment.
- Western blot and qRT-PCR were used to measure protein and gene expression related to drug resistance, autophagy, and the PI3K/Akt/mTOR pathway.
- In vivo studies using a mouse tumor xenograft model to evaluate CRNDE's effect on tumor growth and ABCG2 expression.
Main Results:
- lncRNA CRNDE was upregulated in TMZ-resistant GBM patients and served as a poor prognostic factor.
- Knockdown of CRNDE significantly reduced glioma cell viability and proliferation while increasing apoptosis, enhancing TMZ chemosensitivity in vitro and in vivo.
- CRNDE knockdown inhibited autophagy by affecting key autophagy-related proteins (LC3 II/I, Beclin1, Atg5, p62) and activating the PI3K/Akt/mTOR pathway.
- CRNDE expression was highly correlated with ABCG2 expression, a key drug-resistance protein.
Conclusions:
- lncRNA CRNDE is a reliable clinical predictor of outcome and prognosis in GBM patients.
- CRNDE acts as a potential biomarker for predicting TMZ treatment response by modulating autophagy and ABCG2 expression via the PI3K/Akt/mTOR pathway.
- Targeting CRNDE may represent a novel therapeutic strategy to overcome TMZ resistance in GBM.
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