Tumorigenesis involves multistep genetic alterations. To elucidate the microRNA (miRNA)–gene interaction network in carcinogenesis, we examined their genome‐wide expression profiles in 96 pairs of tumor/non‐tumor tissues from hepatocellular carcinoma (HCC). Comprehensive analysis of the coordinate expression of miRNAs and mRNAs reveals that miR‐122 is under‐expressed in HCC and that increased expression of miR‐122 seed‐matched genes leads to a loss of mitochondrial metabolic function. Furthermore, the miR‐122 secondary targets, which decrease in expression, are good prognostic markers for HCC. Transcriptome profiling data from additional 180 HCC and 40 liver cirrhotic patients in the same cohort were used to confirm the anti‐correlation of miR‐122 primary and secondary target gene sets. The HCC findings can be recapitulated in mouse liver by silencing miR‐122 with antagomir treatment followed by gene‐expression microarray analysis. In vitro miR‐122 data further provided a direct link between induction of miR‐122‐controlled genes and impairment of mitochondrial metabolism. In conclusion, miR‐122 regulates mitochondrial metabolism and its loss may be detrimental to sustaining critical liver function and contribute to morbidity and mortality of liver cancer patients.