Comparison of gene expression signature obtained from systems biology transcriptome data analysis of peripheral blood mononuclear cells in the patients with HCC, CHB and healthy control

Abstract

Hepatitis B virus (HBV) causes acute and chronic hepatitis B (CHB) in humans. CHB is the leading cause of liver failure, cirrhosis, and hepatocellular carcinoma (HCC) worldwide. Despite effective drugs and vaccines, HBV infection remains a major health concern. Therefore, it is necessary to discover new diagnostic and therapeutic methods. Using a minimally invasive method such as blood sampling can be an attractive alternative to a liver biopsy. Peripheral blood mononuclear cells (PBMCs) are important components of the immune system, and the examination of key genes in them has shown the relationship between differential gene expression and diseases, and this issue can be used as a clinical tool. Object: aim of this study is Evaluation of how to change the transcription profile in PBMCs from HCC and CHB patients. These differential expression profiles may be useful for prognostication and staging in different phases of hepatitis B disease. Materials and Methods: In this study, a total of 60 whole blood samples from the participants were collected, including those from healthy people (20 people), CHB patients (20 people) (samples from healthy people and CHB were received from the central laboratory of East Azerbaijan province), and primary HCC (20 people) were received fromfrom the liver transplantation department of Imam Khomeini Hospital in Tehran. Demographic information was obtained from all patients. Patients with immunosuppression, hepatitis C virus (HCV), hepatitis D virus (HDV) positivity, and people with gastrointestinal and liver diseases were excluded from the study. Informed consent was obtained from each participant, and the study was approved by the Ethics Committee of Tabriz University of Medical Sciences. Immediately after blood collection, RNA was extracted from fresh PBMC samples using TRIzol solution. Microarray gene expression data were extracted from GEO gene expression database and the differentially expressed genes (DEGs) of each sample were analyzed by GEO2R server. Enrichment of genes was done (by DAVID Functional Annotation Tool server). Cytoscape software was used to construct protein-protein interaction networks (PPIs) and measure topology characteristics. Finally, SYBR-Green Real-Time PCR was used to confirm their expression changes. In addition, the viral load was also determined in the serum. Results: Comparing the gene expression profiles of healthy people and CHB patients showed that there are 238 DEGs (145 genes with increased expression and 93 genes with decreased expression). Also, 88 DEGs were obtained in the comparison of gene expression profiles of CHB and HCC patients, so 48 genes had increased expression and 40 genes had decreased expression. The four genes (MAX, EGR1, JUN, and MAFF) were introduced as hub genes. Also, the HBV load in HCC patients was significantly higher than that in CHB patients.