Entamoeba histolytica is a human pathogen, which can survive using haemoglobin (Hb) as only iron supply. Two probable haemophores (Ehhmbp26 and Ehhmbp45) are involved in iron acquisition in this parasite. However, mechanisms related to their transcriptional regulation have not been studied yet. In the present work, transcriptional profiles of both genes were evaluated in trophozoites cultures grown with different iron sources. ehhmbp26 gene was repressed totally by free iron, whereas ehhmbp45 gene showed clearly detectable mRNA levels. Expression profiles for both genes were significantly increased under iron privation condition. Interestingly, ehhmbp26 transcript was highly expressed by Holo-transferrin presence. This induction appears to be independent of direct contact between these proteins, because, in vitro assays evidenced that Ehhmbp26 protein was unable to bind this metalloprotein. Besides, in silico analysis of promoter nucleotide sequences of ehhmbp26 and ehhmbp45 genes revealed some distinctive core promoter elements described in E. histolytica and T-rich regions. Taking altogether these data suggest in E. histolytica dissimilar transcriptional mechanisms involved on iron acquisition control the expression of these genes, and they are unlike to those previously described for instance: in bacteria. Our findings evidenced this pathogen regulates the expression of ehhmbp26 and ehhmbp45 genes depending on the available iron supply, always ensuring the success of its infective process.