Liver X Receptors (LXRs) belong to the nuclear receptor’s superfamily of ligand-activated transcription factors, whose endogenous agonists are oxysterols. They play a key role in the regulation of cholesterol homeostasis, induce the de novo synthesis of triacylglycerides, and counteract pro-inflammatory effects. LXRs are also known to compromise cell proliferation in several cancer models. However, their role in breast cancer (BC) has not been depth studied. Here, we have examined the potential involvement of LXRs in BC cells with special emphasis on their possible crosstalk with the Estrogen Receptor alpha (ERɑ). We performed colony formation (CFA) and propidium iodide staining assays in MCF-7 cells treated with or without the ERɑ agonist, Estradiol (E2) and the LXR synthetic agonist, GW3965. Our results showed that GW3965 impaired the cell proliferation capacity induced by E2. To understand the functional pathways involved in these effects, we performed bulk RNA-seq experiments. The differentially expressed genes between E2 and E2+GW3965 conditions revealed several genes whose expression was affected by GW3965, which are widely enriched in terms associated to DNA replication, cell cycle checkpoint signaling and transition between the cell cycle stages (padj<0.05, DESeq2 and clusterProfiler) including genes such as E2F1, E2F7, PCNA, BRCA1, CCNE2, CDK2, CDC7, MKI67, EEIG1 and the MCM family. In addition, we studied the dynamics of the nuclear organization of the Liver X Receptor in LXR-GFP transfected MCF-7 live cells using super-resolution microscopy (Airyscan). We found that the combined treatment (E2+GW3965) led to a decrease in the number of nuclear condensates (foci) of LXR, compared to the GW3965 treatment alone. These results combined show that the LXR interferes with Estrogen Receptor-dependent genomic regulation in MCF7 cells, contributing to the idea of an interaction between these two receptors.