Evaluation and Molecular Expression of miRNAs miRNA21 an miRNA122 in Hepatocellular Carcinoma

Abstract

Hepatocellular carcinoma is most important reason of cancer deaths among almost 80% of total liver cancers and HCV infection is foremost risk factor. The expression pattern and function of micro-RNA involved in HCC progression provides new insights for disease pathogenesis. Current research work is basically involves wet lab and in silico approaches. Wet lab research works carried out for evaluation of expression pattern of miR-122 and miR-21 in the HCV mediated genesis of HCC using the Real-Time PCR expression system. Relative quantification of mi-RNAs were performed by 2^-and#916;and#916;CT method with U6 mi-RNA as internal reference. Expression of miR-122 was found 7.9 times upregulated in HCV positive diseased group G2 in comparison to healthy control group G1 while miR-21 gene expression was observed 17.4 times higher in HCV positive diseased group G2 in comparison to healthy control group G1. The in silico experiment were performed for the identification of liver specific oncogenic micro-RNAs (mir-21, miR-155, mir-221 and miR-224) involved in the genesis and progression of HCV mediated HCC genesis. Retrieved nucleotide FASTA sequences from GenBank were used as input for RNAfold server and as output result provides secondary structure. 3D-molecular structure was prepared with RNAComposer modeling server and model quality assessed by RNA tools online server utilizes RASP and Dfire methods. Prepared 3D structural model was docked with ligand LHPP (Phospholysine Phosphohistidine Inorganic pyrophosphate Phosphatase) by HDOCK server. Docking score obtained for docking ligand LHPP with miR-21, miR-155, miR-221 and miR-224 is -298.13, -291.00, -309.65 and -286.13 respectively. Overall research result demonstrated that expression pattern of miR-122 and miR-21 could be used in the assessment of HCV patients as a serum diagnostic marker leads to development of HCC. Generated molecular model could be used by researcher for better structural knowledge of mi-RNAs and molecular docking of an anti-cancerous compound for the prevention of HCC. newline