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Dilek Cansu Gurer1, İpek Erdogan1, Ulvi Ahmadov1
1Noncoding RNA Laboratory, Department of Molecular Biology and Genetics, Ízmir, Institute of Technology, Izmir, Turkey.
Abstract:
Cisplatin is a well-known cancer chemotherapeutic agent but how extensively long non-coding RNA (lncRNA) expression is modulated by cisplatin is unknown. It is imperative to employ a comprehensive approach to obtain a better account of cisplatin-mediated changes in the expression of lncRNAs. In this study, we used a transcriptomics approach to profile lncRNAs in cisplatin-treated HeLa cells, which resulted in identification of 10,214 differentially expressed lncRNAs, of which 2,500 were antisense lncRNAs. For functional analyses, we knocked down one of the cisplatin inducible lncRNAs, death receptor 5 antisense (DR5-AS) lncRNA, which resulted in a morphological change in HeLa cell shape without inducing any cell death. A second round of transcriptomics-based profiling revealed differential expression of genes associated with immune system, motility and cell cycle in DR5-AS knockdown HeLa cells. Cellular analyses showed that DR5-AS reduced cell proliferation and caused a cell cycle arrest at S and G2/M phases. Moreover, DR5-AS knockdown reduced the invasive capacity of HeLa cells in zebrafish xenograft model. These results suggest that cisplatin-mediated pleiotropic effects, such as reduction in cell proliferation, metastasis and cell cycle arrest, may be mediated by lncRNAs.
Insights
Cisplatin cancer therapy affects long non-coding RNA (lncRNA) expression. This study identified cisplatin-responsive lncRNAs, including DR5-AS, which regulates cell proliferation, cell cycle, and invasion, suggesting lncRNAs mediate cisplatin
Area of Science:
- Molecular Biology
- Genomics
- Cancer Research
Background:
- Cisplatin is a widely used chemotherapy drug.
- The impact of cisplatin on long non-coding RNA (lncRNA) expression is not fully understood.
- Comprehensive profiling of lncRNAs is necessary to understand cisplatin's effects.
Purpose of the Study:
- To comprehensively profile lncRNA expression changes induced by cisplatin in HeLa cells.
- To investigate the functional role of a specific cisplatin-inducible lncRNA, DR5-AS.
- To elucidate the involvement of lncRNAs in mediating cisplatin's pleiotropic effects.
Main Methods:
- Transcriptomics-based profiling of lncRNAs in cisplatin-treated HeLa cells.
- Knockdown of the death receptor 5 antisense (DR5-AS) lncRNA.
- Second transcriptomics profiling of DR5-AS knockdown cells.
- Cellular assays including proliferation, cell cycle analysis, and invasion assays in a zebrafish xenograft model.
Main Results:
- Identification of 10,214 differentially expressed lncRNAs, including 2,500 antisense lncRNAs, in cisplatin-treated HeLa cells.
- Knockdown of DR5-AS altered HeLa cell morphology, reduced cell proliferation, induced S and G2/M phase cell cycle arrest, and decreased invasive capacity.
- Genes related to the immune system, motility, and cell cycle were differentially expressed in DR5-AS knockdown cells.
Conclusions:
- Cisplatin significantly modulates lncRNA expression in HeLa cells.
- The lncRNA DR5-AS plays a role in regulating cell proliferation, cell cycle progression, and invasion.
- lncRNAs may mediate several of cisplatin's therapeutic effects, including reduced proliferation and metastasis.
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