The Expression of AIP -Related Molecules in Elucidation of Cellular Pathways in Pituitary Adenomas

Germline mutations in the aryl hydrocarbon receptor interacting protein ( AIP ) gene predispose to the development of pituitary adenomas. Here, we characterized AIP mutation positive ( AIP mut+) and AIP mutation negative ( AIP mut−) pituitary adenomas by immunohistochemistry. The expressions of the AIP-related proteins aryl hydrocarbon receptor (AHR), AHR nuclear translocator (ARNT), cyclin-dependent kinase inhibitor 1B encoding p27(Kip1), and hypoxia-inducible factor 1-α were examined in 14 AIP mut+ and 53 AIP mut− pituitary adenomas to detect possible expression differences. In addition, the expression of CD34, an endothelial and hematopoietic stem cell marker, was analyzed. We found ARNT to be less frequently expressed in AIP mut+ pituitary adenomas ( P = 0.001), suggesting that AIP regulates the ARNT levels. AIP small interfering RNA-treated HeLa, HEK293, or Aip -null mouse embryonic fibroblast cells did not show lowered expression of ARNT. Instead, in the pituitary adenoma cell line GH3, Aip silencing caused a partial reduction of Arnt and a clear increase in cell proliferation. We also observed a trend for increased expression of nuclear AHR in AIP mut+ samples, although the difference was not statistically significant ( P = 0.06). The expressions of p27(Kip1), hypoxia-inducible factor 1-α, or CD34 did not differ between tumor types. The present study shows that the expression of ARNT protein is significantly reduced in AIP mut+ tumors. We suggest that the down-regulation of ARNT may be connected to an imbalance in AHR/ARNT complex formation arising from aberrant cAMP signaling.