Long Non-Coding RNA HAND2-AS1 Acts as a Tumor Suppressor in High-Grade Serous Ovarian Carcinoma
Abstract
:1. Introduction
2. Results
2.1. HAND2-AS1 as an Important Long Non-Coding RNA (lncRNA) in High-Grade Serous Carcinoma (HGSC)
2.2. HAND2-AS1 Expression Is Consistently Downregulated in Ovarian Cancer Cell Lines Possibly Due to Promoter Hypermethylation
2.3. Expression of HAND2-AS1 in Ovarian Cancer Cells Decreases Their Adhesion to Extracellular Matrix, Migration, and Viability
2.4. HAND2-AS1 Expression Is Rescued after Treatment with HDAC Inhibitor
2.5. Identification of the HAND2-AS1-Associated mRNA and ceRNA Network along with Enriched Gene Ontology Biological Processes and Pathways in HGSC
3. Discussion
4. Materials and Methods
4.1. Differential Analysis of Coding and Long Non-Coding RNAs
4.2. Expression of HAND2-AS1 in Various Cancers
4.3. Venn Diagrams
4.4. Cell Lines and Culture Methods
4.5. Treatment with AZA
4.6. Plasmid Preparation
4.7. RNA, cDNA, and qRT-PCR
- ABL fwd 5′-TGGAGATAACACTCTAAGCATAACTAAAGG-3′
- ABL rev 5′-GATGTAGTTGCTTGGGACCCA-3′
- HAND2-AS1 fwd 5′-GTCTACGACTGCCTGTGACT-3′
- HAND2-AS1 rev 5′-TGTCCCATATACAAGAATCGACA-3′
- PAX8 fwd 5′-CCCTTCCAACACGCCACT-3′
- PAX8 rev 5′-CTGCTTTATGGCGAAGGGTG-3′
4.8. Migration Assay
4.9. Adhesion Assay
4.10. Viability Assay
4.11. HDAC Inhibitor Treatment
4.12. mRNA Network
4.13. HAND2-AS1 ceRNA Network Prediction
4.14. Figure for Graphical Summary
4.15. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
lncRNA | Long non-coding RNA |
ncRNA | Non-coding RNA |
HGSC | High-Grade Serous Carcinoma |
FT | Fallopian Tube |
OC | Ovarian Cancer |
HAND2-AS1 | HAND2 Anti sense RNA 1 |
5-AZA | 5-Aza-2’–deoxycytidine |
mRNA | Messenger RNA |
ceRNA | Competing endogenous RNA |
HDAC | Histone De-Acetylases |
miRNA | Micro RNA |
GO BP | Gene Ontology Biological Process |
KEGG | Kyoto Encyclopedia of Genes and Genomes |
TCGA-OV | The Cancer Genome Atlas – Ovarian Cancer |
GTEx | Genotype Tissue Expression |
GEPIA | Gene Expression Profiling Interactive Analysis |
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Gokulnath, P.; de Cristofaro, T.; Manipur, I.; Di Palma, T.; Soriano, A.A.; Guarracino, M.R.; Zannini, M. Long Non-Coding RNA HAND2-AS1 Acts as a Tumor Suppressor in High-Grade Serous Ovarian Carcinoma. Int. J. Mol. Sci. 2020, 21, 4059. https://doi.org/10.3390/ijms21114059
Gokulnath P, de Cristofaro T, Manipur I, Di Palma T, Soriano AA, Guarracino MR, Zannini M. Long Non-Coding RNA HAND2-AS1 Acts as a Tumor Suppressor in High-Grade Serous Ovarian Carcinoma. International Journal of Molecular Sciences. 2020; 21(11):4059. https://doi.org/10.3390/ijms21114059
Chicago/Turabian StyleGokulnath, Priyanka, Tiziana de Cristofaro, Ichcha Manipur, Tina Di Palma, Amata Amy Soriano, Mario Rosario Guarracino, and Mariastella Zannini. 2020. "Long Non-Coding RNA HAND2-AS1 Acts as a Tumor Suppressor in High-Grade Serous Ovarian Carcinoma" International Journal of Molecular Sciences 21, no. 11: 4059. https://doi.org/10.3390/ijms21114059