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Integrated in-silico and in-vitro analysis of lncRNA H19/miRNA-675/p53 in OSCC: Structural characterization and
Ramya Sekar1, Selvaraj Jayaraman2, Vishnupriya Veeraraghavan3
1Department of Oral & Maxillofacial Pathology and Oral Microbiology, Meenakshi Ammal Dental College and Hospital, Meenakshi Academy of Higher Education and Research (Deemed to Be University), Alapakkam Main Road, Maduravoyal, Chennai-95, India.
Background:
Long non-coding RNAs (lncRNA) H19 has drawn special attention because of its varied role in several malignancies, including OSCC. Therefore, this study was conducted to assess the association between H19-miR675-p53 by in-silico analysis, quantify the expression levels of H19, miRNA-675, and target oncogene p53 in cancerous versus normal individuals, and Correlate the Clinicopathological findings with their expression pattern.
Methods:
The secondary structure of lncRNA H19 was predicted using the RNAfold web server ( http://rna.tbi.univie.ac.at/cgi-bin/RNAWebSuite/RNAfold.cgi ). The FASTA sequence of H19 was retrieved from the NCBI database ( https://www.ncbi.nlm.nih.gov/ ). We performed molecular docking studies to analyze the interaction between miRNA-675 and p53 using the MDockPP ( https://zougrouptoolkit.missouri.edu/MDockPP/ ) web server. Real-time PCR was used to measure the amounts of H19 and miR-675, and Immunohistochemistry was used to analyse the pattern of p53 expression.
Result:
The study successfully associated miR-675 from the first exon of H19 modulating p53 via in silico analysis. It was found that H19 and miR-675 levels were higher in OSCC patients (3.12 ± 1.16) compared to healthy patients (1.0 ± 0.0), and was statistically significant (p-value < 0.001).
Conclusion:
The specificity of H19 expression in OSCC compared to normal presents an attractive target for cancer-specific therapies, minimizing the risk of off-target effects.
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