Screening of the HFE gene mutations in patients with idiopathic cirrhosis by PCR-RFLP technique

SCREENING OF THE HFE GENE MUTATIONS IN PATIENTS WITHIDIOPATHIC CIRRHOSIS BY PCR-RFLP TECHNIQUESaffet ÖZTÜRKCirrhosis that is one of the important disease of the liver results from externaland internal etiologic factors. Liver cirrhosis is classified according to anatomy,etiology, activity, prognose and stage. Cirrhosis, excluded from the etiologicalfactors and defined very well, is called idiopathic cirrhosis and constitutes 5-10 % ofcirrhosis patients. Iron overload is the most frequent risk factor in the liver. Severalgenetic factors are involved in the iron metabolism. One of them is the HFE gene,and HFE gene (for hemochromatosis) mutations cause iron overload and so thataffect function of the liver negatively.The aim of our project, is to investigate the mutations (C282Y, H63D, S65C)in the HFE gene in patients with idiopathic cirrhosis using PCR-RFLP, and todetermine the possible relation between HFE gene mutations and the disease. HFEgene mutations were investigated with PCR-RFLP technique. After DNA isolationfrom peripheral blood, the DNA regions including every three mutations wereamplified with PCR technique. PCR products were digested with spesific restrictionenzymes for mutation sites. Digested fragments were run on the electrophoresis, andgenotypes were determined according to band patterns.One of the 16 idiopathic cirrhosis patients is heterozygous and one of them isalso homozygous for H63D mutation. In 141 healthy control group, 30 and 2 of themwere determined heterozygous and homozygous, respectively. C282Y and H63Dmutations were not found in both the patients and the control group. When genotypesand biochemical iron parameters (serum iron concentration, ferritin, total ironbinding capacity and transferrin saturation) of the patients and the control individualswere compared, the ferritin, serum iron concentration and transferrin saturationvalues of these subjects who were homozygous for the H63D mutation were higherthan the heterozygous and normal genotypes for the this mutation. It was discussedwhether the HFE gene mutations (C282Y, H63D and S65C) are the etiologic factorswhich cause the idiopathic cirrhosis.Key words: Idiopathic cirrhosis, HFE gene, PCR, RFLP.