Molecular characterization of aromatase inhibitor-resistant, tamoxifen-resistant and LTEDaro cell lines.

To determine potential genes involved in mediating resistance to aromatase inhibitors (AIs), a microarray study was performed using MCF-7aro (aromatase overexpressing) cells that are resistant to letrozole (T+LET R), anastrozole (T+ANA R) and exemestane (T+EXE R), as well as LTEDaro and tamoxifen-resistant (T+TAM R) lines for comparison. Based on hierarchical clustering, estrogen-responsive genes were found to be differentially expressed in AI-resistant lines versus LTEDaro and T+TAM R. Additional genome-wide analysis showed that gene expression profiles of the non-steroidal AI-resistant lines were most closely correlated and that T+EXE R lines exhibit differing profiles. Also, LTEDaro and T+TAM R lines are inherently different from expression profiles of AI-resistant lines. Further characterization of these resistant lines revealed that T+LET R, T+ANA R and LTEDaro cells contain a constitutively active estrogen receptor alpha (ERalpha) that does not require the ligand estrogen for activation. Ligand-independent activation of ERalpha does not activate identical estrogen-responsive gene profiles in AI-resistant lines as in LTEDaro lines, thereby establishing differing mechanisms of resistance. This ligand-independent activation of ER was not observed in the parental cell lines MCF-7aro, T+EXE R or T+TAM R cells. Based on the steroidal structure of EXE, our laboratory has shown that this AI has weak estrogen-like properties, and that EXE resistance involves an ER-dependent crosstalk with EGFR growth factor signaling. Recent studies in our laboratory pertaining to pre-clinical models of AI treatment revealed that intermittent use of EXE delays the onset of acquired resistance in comparison to continuous treatment. Specific molecular mechanisms involved in intermittent use of EXE are currently being explored, based on microarray gene expression profiling. Lastly, our laboratory has initiated a study of microRNAs and their potential role in regulating target genes involved in AI-resistance. Overall, we propose a model of acquired resistance that progresses from hormone-dependence (T+TAM R and T+EXE R) to hormone-independence (T+LET R and T+ANA R), eventually resulting in hormone-independence that does not rely on conventional ER signaling (LTEDaro).