Hemochromatosis is an autosomal recessive iron overload disorder characterized by excessive iron accumulation, which can lead to various health complications, including liver fibrosis, cirrhosis, and an increased risk of hepatocellular carcinoma (HCC). Iron overload promotes oxidative stress, DNA damage, and cellular proliferation, all of which are implicated in carcinogenesis. The TFR2 gene variant rs7385804 has been implicated in hemochromatosis; however, its role in the Pakistani population and its potential link to cancer progression remain unexplored. This study aimed to investigate the association between the TFR2 gene variant rs7385804 and hemochromatosis in Pakistani individuals and assess its possible role in cancer-related mechanisms. We employed a case--control study design, recruiting 200 hemochromatosis patients and 200 healthy controls from the Pakistani population. Genomic DNA was extracted from peripheral blood samples using the phenol-chloroform method. Sanger sequencing and Tetra-ARMS PCR were used to detect the TFR2 gene variant rs7385804. Our results demonstrated a significant association between the TFR2 gene variant rs7385804 and hemochromatosis in the Pakistani population (p < 0.001). The frequency of the variant allele was higher in patients (35%) compared to controls (15%). Sanger sequencing confirmed the presence of the variant in 70% of patients, while Tetra-ARMS PCR showed 90% concordance with sequencing results. Iron overload, mediated by TFR2 dysregulation, can lead to oxidative stress and genomic instability, key drivers in the development of hepatocellular carcinoma and other malignancies. This study highlights the utility of Sanger sequencing and Tetra-ARMS PCR in detecting the TFR2 gene variant rs7385804 and provides evidence that this variant may serve as a genetic marker for hemochromatosis and a potential contributor to iron-related carcinogenesis in the Pakistani population. Further studies are required to validate these findings and elucidate the clinical implications of this association in cancer development.