Background: Hepatocellular carcinoma (HCC) poses a substantial global health burden characterized by high mortality rates and limited treatment options, particularly in regions with high prevalence rates such as Egypt. To investigate the role of miR-16 targeting HO-1 in Egyptian patients with liver cancer, elucidating the interplay between miRNAs and their target genes in hepatocarcinogenesis, and assessing their prognostic significance and therapeutic potential in this high-risk population. Methods: Fifty patients with HCC undergoing follow-up at the general hospital of Benha University, were included in this study. Tumoral and healthy marginal tissues were collected, and RNA was extracted and subjected to reverse transcription to synthesize complementary DNA (cDNA). The expression levels of miR-16 and HO-1 were determined using quantitative real-time PCR, and correlations with clinical parameters were assessed. Results: Analysis of demographic data revealed a higher proportion of patients over 60 years old, with a majority being female. Significant alterations in the expression levels of HO-1 and miR-16 were observed in tumor samples compared to controls, suggesting their involvement in HCC pathogenesis. Differential expression patterns between metastatic and non-metastatic samples indicate potential roles in metastasis initiation and progression. Correlation analysis revealed associations between genetic expression, demographic characteristics, and clinical parameters, underscoring the complex interplay between miR-16, HO-1, and disease progression. Conclusion: The findings highlight the potential of miR-16 targeting HO-1 as a therapeutic strategy and emphasize the importance of personalized treatment approaches in high-risk populations. Further research is warranted to elucidate the precise mechanisms and develop novel therapeutic interventions for liver cancer