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[Dysregulation of Long Non-coding RNAs in Glioblastoma Multiforme and Their Study Through Use of Modern
Background:
Glioblastoma (GBM) is the most frequent primary brain tumor characterized by an unfavourable prognosis despite multimodal therapy. Therefore, a lot of efforts and financial resources are dedicated to the research of new therapeutic targets and prognostic or predictive biomarkers. Long non-coding RNAs (lncRNAs) are regulators of gene expression which play a significant role in GBM pathology and, thus, present promising candidates.
Material And Methods:
Our study included 14 patients with GBM and 8 patients with intractable epilepsy from whom we acquired brain tissues during surgical intervention. Ribosomal RNA depleted RNA was used for sequencing by NextSeq 500 instrument (Illumina). Statistical analysis evaluated 24,087 protein-coding and 8,414 non-coding RNAs and their sequential variants with non-zero reads per kilobase per million mapped reads (RPKM) at least in one sample. CLC Genomic Workbench was used for the alignment and target counts. Targeted downregulation of up-regulated ZFAS1, one of the identified lncRNA, level has been carried out by the transient transfection of specific small interfering RNA (siRNA) in GBM stable cell lines (A172, U87MG, T98G). The success of transfection and viability were analyzed in vitro using quantitative real time polymerase chain reaction and MTT assay, resp.
Results:
Statistical analysis has revealed 274 (p < 0.01) dysregulated lncRNAs in GBMs in comparison with non-tumor brain tissues. Moreover, the results have showed 489 dysregulated mRNAs (p < 0.0001) and 26 mRNAs (p < 0.000001). Transfection of ZFAS1 inhibitor led to successful downregulation of ZFAS1 expression level, although it did not have a significant effect on proliferation of GBM cells.
Conclusion:
We described a significant dysregulation of lncRNAs and mRNAs in GBM tissue in comparison with non-tumor tissue. We also succesfully decreased expression level of ZFAS1, which in turn, however, had no impact on the viability of GBM cell lines.Key words: glioblastoma - long non-coding RNA - next-generation sequencing The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers. This tudy was supported by Ministry of Health of the Czech Republic, grant No. 15-33158A. All rights reserved.Submitted: 19. 3. 2018Accepted: 10. 4. 2018.
Insights
Glioblastoma (GBM) exhibits significant alterations in long non-coding RNAs (lncRNAs) and messenger RNAs (mRNAs). While ZFAS1 (a lncRNA) was successfully downregulated, it did not affect GBM cell viability.
Area of Science:
- Oncology
- Genomics
- Molecular Biology
Background:
- Glioblastoma (GBM) is an aggressive brain tumor with poor prognosis.
- Identifying novel therapeutic targets and biomarkers is crucial for GBM treatment.
- Long non-coding RNAs (lncRNAs) are implicated in GBM pathogenesis and represent potential targets.
Purpose of the Study:
- To investigate the differential expression of lncRNAs and mRNAs in GBM.
- To identify potential lncRNA biomarkers for GBM.
- To evaluate the therapeutic potential of targeting ZFAS1 in GBM.
Main Methods:
- Next-generation sequencing (NGS) of RNA from GBM and non-tumor brain tissues.
- Bioinformatic analysis to identify dysregulated lncRNAs and mRNAs.
- In vitro experiments using small interfering RNA (siRNA) to downregulate ZFAS1 in GBM cell lines.
Main Results:
- 274 lncRNAs and 489 mRNAs were found to be significantly dysregulated in GBM.
- ZFAS1, an upregulated lncRNA, was successfully downregulated using siRNA.
- Downregulation of ZFAS1 did not significantly impact the proliferation of GBM cell lines.
Conclusions:
- GBM tissues show significant dysregulation of lncRNAs and mRNAs compared to non-tumor tissues.
- Targeting ZFAS1 may not be a viable therapeutic strategy for GBM due to its lack of effect on cell viability.
- Further research is needed to explore other dysregulated lncRNAs as potential therapeutic targets in GBM.
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