CIBTech Journal of Biotechnology ISSN: 2319–3859 (Online) An Online International Journal Available at http://www.cibtech.org/cjb.htm 2012 Vol. 1 (2&3) Jul-Sept. &Oct.-Dec., pp.22-27/Mistry et al. Research Article 22 MUTATIONAL ANALYSIS OF HFE GENE IN PREMENOPAUSAL AND MENOPAUSAL WOMEN IN POPULATION OF GUJARAT * Kinnari Mistry, Sanjay Lal, Vinni Malhotra and Ekta Rana Department of Medical Biotechnology, Ashok and Rita Patel Institute of Integrated Study & Research in Biotechnology and Allied sciences (ARIBAS), New Vallabh Vidyanagar – 388121 (Gujarat) India * Author for Correspondence ABSTRACT Heriditary hemachromatosis(HH) is the prototype disease for primary iron overload. The gene that causes most cases of HH is designated as HFE. Two missense mutations (C282Yand H63D) of this gene are found to be associated with HH phenotype. In present study, our aim is to find out HFE gene mutations in premenopausal and menopausal women in population of Gujarat. Polymerase chain reaction-restriction fragment length polymorphism method was used for screening C282Yand H63D mutation in 54 subjects [Premenopausal phase iron-overload subjects (27); Menopausal phase iron-overload subjects (27)] and 21 healthy controls. Our findings show that out of 54 female iron-overload subjects, 1 had a heterozygous mutation of the H63D region of HFE gene. The genotype frequency of menopausal phase female subjects was 24(89 %) for homozygous H63D and 3(11.11%) for heterozygous H63D. This corresponds to allelic frequency of 94.44% for C-allele. The second important finding of our study was that all subjects were free from C282Y mutation. The results of our case-control study indicate that H63D has a positive association with iron-overload. We did not find any C282Y mutation among the women who participated in this study. Ideally, a sample larger than ours should be studied in a genetic association study to rule out the chance factor. Key Words: HFE gene, Primary Iron Overload, Heriditary Hemachromatosis, HFE Mutations INTRODUCTION Iron is indispensable for basic cellular functions. However, this metal is also a catalyst for chemical reactions associated with the production of reactive oxygen species, which may lead to oxidative stress and cellular damage. Hence, the controlled regulation of iron homeostasis is necessary to keep the body iron at a moderate level to avoid iron deficiency and iron overload (Parkkila et al., 2001). Body iron homeostasis is regulated primarily by duodenal and upper small intestinal absorption and is responsive to body iron stores. Hence, iron absorption is increased during iron deficiency and down-regulated when iron are replete (Pietrangelo et al., 1995). There is no effective method for the elimination of excess body iron, and iron overload from iatrogenic or idiopathic pathological causes can lead to multiple systemic complications. Hereditary haemochromatosis, the prototype disorder of iron overload due to misregulated iron homeostasis in humans, is caused by an inappropriate increase in iron absorption in the duodenum and upper small intestine. Iron overload increases the risk of various clinical complications such as arthritis, diabetes, cardiomyopathy, skin pigmentation, gonadal failure and liver cirrhosis (CLD) (Fleming et al., 2002). The genes identified to be responsible for primary iron overload in HH are HFE, HJV, HAMP, TFR2 and SLC11A (Feder et al., 1996). The hereditary hemochromatosis gene HFE plays a pivotal role in iron homeostasis. The hereditary hemochromatosis gene HFE (6p21.3), 4 Mb telomeric to the HLA-A locus, and its product has a structure similar to MHC class I molecules. It has a critical role in iron homeostasis. Two missense mutations (C282Yand H63D) of this gene are found to be associated with HH phenotype (Feder et al., 1996). The C282Y mutation, most frequent amongst Caucasians (Beutler et al., 1996) results from a G-to-A transition at nucleotide 845 of the HFE gene (845 G → A) that produces a substitution of cysteine for a tyrosine at amino acid position 282 (C282Y) in the protein product. In the