With the aim of evaluating the relationship between pituitary tumorigenesis and the presence of estrogen receptor-α (ERα) by immunohistochemistry (IH) and their relevance to patients’ clinical presentation, hormonal phenotypes of adenomas, preoperative neuroimaging findings, and the index of cellular replication MIB-1, a study was conducted with material from 91 women and 67 men with pituitary adenomas. The patients had acromegaly (29.7%), Cushing’s disease (14.6%), hyperprolactinemic syndrome (20.9%), and clinically nonfunctioning tumors (34.8%). Of the patients, 14.6% had microadenomas, 52.5% had macroadenomas with or without suprasellar growth, 28.5% had invasive macroadenomas and in 4.4% the adenoma was not visualized. IH showed that 43 were positive for growth hormone (GH), 16 for corticotropin (ACTH), 18 for prolactin (PRL), 18 for PRL+GH, 6 for luteinizing hormone (LH) and follicle-stimulating hormone (FSH), 15 had a plurihormonal reaction, and 42 had nonfunctioning adenomas. The presence of ERα was positive in 9/158 adenomas with a median value for the percentage of labeled cells of 42.89%, and in 6/16 controls (autopsy samples) with a median value for the percentage of labeled cells of 0.024%. ERα was significantly more prevalent in controls than in patients with adenomas (37.5 versus5.7%; p = 0.001); however, the mean ERα concentration in adenomas was significantly greater than in controls (42.89 versus 0.024%; p < 0.001). No significant difference in the concentration of ERα was found across the clinical presentations, hormonal phenotypes or findings of preoperative CT. Among the ERα-positive adenomas, ERα values were significantly greater in invasive macroadenomas (80%) than in microadenomas (3.33%). MIB-1 values did not differ significantly between ERα-positive and -negative adenomas, nor did the correlation between ERα values and the MIB-1 index attain significance in the total sample, even when only ERα-positive adenomas and positive MIB-1 indexes were considered. It was concluded that, when present in pituitary tumors, ERα exhibits a high concentration, and is more common in nonfunctioning and invasive adenomas, but absent in ACTH-secreting ones.