PPARγ signaling correlates with LATS2 in human and mouse tumors and promotes cell death in a LATS2-dependent manner. PPARγ signaling correlates with LATS2.

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PPARγ signaling correlates with LATS2 in human and mouse tumors and promotes cell death in a LATS2-dependent manner. PPARγ signaling correlates with LATS2 in human and mouse tumors and promotes cell death in a LATS2-dependent manner. (A) KEGG pathways significantly enriched within the list of genes commonly down-regulated in Lats2-CKO PyMT (compared with WT-PyMT) tumors and LATS2L lumB human tumors (TCGA). (B) RT-qPCR quantification of Pparg and Plin1 expression in tumors derived from 3 mo old Lats2-CKO PyMT (n = 9) and WT-PyMT littermate controls (n = 6); mean ± SEM; *P-value < 0.05, ***P-value < 0.001. (C) Expression levels of the indicated genes (left) and proteins (right) in cultured WT PyMT and Lats2-CKO PyMT tumor-derived cells stably transduced with LATS2 or vector control; mean ± SD of 2 technical repeats. (D) ZR75-1 cells were transfected with GFP-LATS2 or control GFP plasmid, stained 48 h later with anti-PPARγ and cleaved caspase 3 (CC3) antibodies, and subjected to imaging flow cytometry (ImageStreamX). Only GFP-positive cells with intact nuclei were analyzed. Cells with nuclear localization of the transfected protein were identified by similarity between GFP and DAPI staining. Percentage of cells positively stained for PPARγ in each subpopulation is presented. Representative images are shown at the bottom. BF = bright field. (E) Left panel: scatter plot depicting PPARG and LATS2 expression in luminal tumors in the TCGA-BRCA dataset (n = 613). Right panel: GSEA of genes ranked according to expression fold change between LATS2L tumors and LATSH tumors in the TCGA lumB dataset. (F) Kaplan-Meier analysis of survival probability of luminal breast cancer patients (METABRIC dataset, n = 1139; Cox proportional hazards model) defined according to expression levels of LATS2 and PPARG.(G) RT-qPCR quantification of PPARG and LPL (left) and LATS2 (right) mRNA in ZR75-1 cells treated for 48 h with increasing concentrations of RGZ (0, 10, 25, 50, and 100 μM, respectively); mean ± SD of two technical replicates. (H) ZR75-1/vector (vector) and ZR75-1/LATS2 (LATS2) cells were treated with 100 μM RGZ for 48 h, followed by PI exclusion analysis; mean ± SEM of the ratio between % PI+ cells in each cell type, from three independent experiments. (I) Cell lines derived from WT-PyMT and Lats2-CKO PyMT tumors were treated with 100 μM RGZ for 48 h, followed by PI exclusion analysis; mean ± SEM of the ratio between % PI+ cells in treated versus untreated cells, from two independent experiments. Noa Furth et al. LSA 2018;1:e201800171 © 2018 Furth et al.