Brief Article
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World J Gastroenterol. Aug 21, 2012; 18(31): 4156-4161
Published online Aug 21, 2012. doi: 10.3748/wjg.v18.i31.4156
Intrahepatic expression of genes related to metabotropic receptors in chronic hepatitis
Andrzej Cieśla, Maciej Kuśmider, Agata Faron-Górecka, Marta Dziedzicka-Wasylewska, Monika Bociąga-Jasik, Danuta Owczarek, Irena Ciećko-Michalska, Dorota Cibor, Tomasz Mach
Andrzej Cieśla, Monika Bociąga-Jasik, Danuta Owczarek, Irena Ciećko-Michalska, Dorota Cibor, Tomasz Mach, Department of Gastroenterology, Hepatology and Infectious Diseases, Jagiellonian University Medical College, 31-521 Kraków, Poland
Maciej Kuśmider, Agata Faron-Górecka, Marta Dziedzicka-Wasylewska, Institute of Pharmacology, Polish Academy of Sciences, 31-343 Kraków, Poland
Author contributions: Cieśla A, Kuśmider M, Faron-Górecka A, Dziedzicka-Wasylewska M, Mach T designed the research; Cieśla A, Kuśmider M, Faron-Górecka A, Dziedzicka-Wasylewska M, Bociąga-Jasik M, Owczarek D, Ciećko-Michalska I, Cibor D performed the study; Kuśmider M, Faron-Górecka A, Dziedzicka-Wasylewska M performed the analytical part of study; Cieśla A, Kuśmider M, Faron-Górecka A, Dziedzicka-Wasylewska M analyzed the data; Cieśla A, Kuśmider M, Faron-Górecka A, Dziedzicka-Wasylewska M, Bociąga-Jasik M, Owczarek D, Ciećko-Michalska I, Cibor D, Mach T wrote the paper.
Correspondence to: Andrzej Cieśla, MD, Department of Gastroenterology, Hepatology and Infectious Diseases, Jagiellonian University Medical College, 31-521 Kraków, Poland. aciesla@su.krakow.pl
Telephone: +48-12-4247340 Fax: +48-12-42473480
Received: September 1, 2011
Revised: April 23, 2012
Accepted: April 27, 2012
Published online: August 21, 2012
Abstract

AIM: To screen for genes related to metabotropic receptors that might be involved in the development of chronic hepatitis.

METHODS: Assessment of 20 genes associated with metabotropic receptors was performed in liver specimens obtained by punch biopsy from 12 patients with autoimmune and chronic hepatitis type B and C. For this purpose, a microarray with low integrity grade and with oligonucleotide DNA probes complementary to target transcripts was used. Evaluation of gene expression was performed in relation to transcript level, correlation between samples and grouping of clinical parameters used in chronic hepatitis assessment. Clinical markers of chronic hepatitis included alanine and aspartate aminotransferase, γ-glutamyltranspeptidase, alkaline phosphatase and cholinesterase activity, levels of iron ions, total cholesterol, triglycerides, albumin, glucose, hemoglobin, platelets, histological analysis of inflammatory and necrotic status, fibrosis according to METAVIR score, steatosis, as well as anthropometric body mass index, waist/hip index, percentage of adipose tissue and liver size in ultrasound examination. Gender, age, concomitant diseases and drugs were also taken into account. Validation of oligonucleotide microarray gene expression results was done with the use of quantitative real-time polymerase chain reaction (qRT-PCR).

RESULTS: The highest (0.002 < P < 0.046) expression among genes encoding main components of metabotropic receptor pathways, such as the α subunit of G-coupled protein, phosphoinositol-dependent protein kinase or arrestin was comparable to that of angiotensinogen synthesized in the liver. Carcinogenesis suppressor genes, such as chemokine ligand 4, transcription factor early growth response protein 1 and lysophosphatidic acid receptor, were characterized by the lowest expression (0.002 < P < 0.046), while the factor potentially triggering hepatic cancer, transcription factor JUN-B, had a 20-fold higher expression. The correlation between expression of genes of protein kinases PDPK1, phosphoinositide 3-kinase and protein kinase A (Spearman’s coefficient range: 0.762-0.769) confirmed a functional link between these enzymes. Gender (P = 0.0046) and inflammation severity, measured by alanine aminotransferase activity (P = 0.035), were characterized by diverse metabotropic receptor gene expression patterns. The Pearson’s coefficient ranging from -0.35 to 0.99 from the results of qRT-PCR and microarray indicated that qRT-PCR had certain limitations as a validation tool for oligonucleotide microarray studies.

CONCLUSION: A microarray-based analysis of hepatocyte metabotropic G-protein-related gene expression can reveal the molecular basis of chronic hepatitis.

Keywords: Metabotropic receptors; Gene expression; DNA oligonucleotides; Quantitative real-time polymerase chain reaction; Chronic hepatitis