Hsa-miR-195 targets PCMT1 in hepatocellular carcinoma that increases tumor life span

Theodor Bilharz Research Institute

Bibliographic Information

Authors: Amer M.; Elhefnawi M.; El-Ahwany E.; Awad A.F.; Gawad N.A.; Zada S.; Tawab F.M.A.

Journal: Tumor Biology

Publisher: Kluwer Academic Publishers

Publication Date: 14 August 2014

Volume / Issue: Volume 35 / Issue 11

Pages: 11301–11309

ISSN: 10104283

DOI: 10.1007/s13277-014-2445-4

Scopus: View on Scopus

PubMed: 25119594

Document Type: Article

Authors and Affiliations

Amer M., Biotechnology Department, Faculty of Biotechnology, Misr University for Science and Technology, Giza, Egypt, Biology Department, American University in Cairo, Cairo, Egypt; Elhefnawi M., Biomedical Informatics and Chemoinformatics Research Group, Center of Excellence for Advanced Sciences, National Research Center, Giza, Egypt; El-Ahwany E., Immunology Department, Theodor Bilharz Research Institute, Giza, Egypt, Biology Department, American University in Cairo, Cairo, Egypt; Awad A.F., Genetics Department, Faculty of Agriculture, Ain-Shams University, Cairo, Egypt; Gawad N.A., Genetics Department, Faculty of Agriculture, Ain-Shams University, Cairo, Egypt; Zada S., Biology Department, American University in Cairo, Cairo, Egypt; Tawab F.M.A., Genetics Department, Faculty of Agriculture, Ain-Shams University, Cairo, Egypt

Abstract

MicroRNAs are small 19–25 nucleotides which have been shown to play important roles in the regulation of gene expression in many organisms. Downregulation or accumulation of miRNAs implies either tumor suppression or oncogenic activation. In this study, differentially expressed hsa-miR-195 in hepatocellular carcinoma (HCC) was identified and analyzed. The prediction was done using a consensus approach of tools. The validation steps were done at two different levels in silico and in vitro. FGF7, GHR, PCMT1, CITED2, PEX5, PEX13, NOVA1, AXIN2, and TSPYL2 were detected with high significant (P < 0.005). These genes are involved in important pathways in cancer like MAPK signaling pathway, Jak-STAT signaling pathways, regulation of actin cytoskeleton, angiogenesis, Wnt signaling pathway, and TGF-beta signaling pathway. In vitro target validation was done for protein-l-isoaspartate (d-aspartate) O-methyltransferase (PCMT1). The co-transfection of pmirGLO-PCMT1 and pEGP-miR-195 showed highly significant results. Firefly luciferase was detected using Lumiscensor and t test analysis was done. Firefly luciferase expression was significantly decreased (P < 0.001) in comparison to the control. The low expression of firefly luciferase validates the method of target prediction that we used in this work by working on PCMT1 as a target for miR-195. Furthermore, the rest of the predicted genes are suspected to be real targets for hsa-miR-195. These target genes control almost all the hallmarks of liver cancer which can be used as therapeutic targets in cancer treatment. © 2014, International Society of Oncology and BioMarkers (ISOBM).

Keywords

Cancer; Luciferase; miRNA; Target prediction; 3' Untranslated Regions; Carcinoma, Hepatocellular; Computer Simulation; Gene Ontology; Humans; Liver Neoplasms; Luciferases; MicroRNAs; Protein D-Aspartate-L-Isoaspartate Methyltransferase; Tumor Cells, Cultured; 3' untranslated region; microRNA; MIRN195 microRNA, human; PCMT1 protein, human; protein dextro aspartate methyltransferase; genetics; human; liver cell carcinoma; liver tumor; metabolism; pathology; tumor cell culture

Citation Information

Scopus Citations: 43

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