Supplementary Figures 1 - 11 from A Genetic Mouse Model of Invasive Endometrial Cancer Driven by Concurrent Loss of Pten and Lkb1 Is Highly Responsive to mTOR Inhibition
Supplementary Figure 1. Representative images of metastases found in the liver (left panel) and peritoneal wall (right panel) of Pten loxp/loxp Lkb1loxp/loxp female mice bearing Pten Lkb1-deficient endometrial tumors. Supplementary Figure 2. Genotyping of endometrial tumors for the recombined alleles of Pten (A) and Lkb1 (B). Genomic DNAs from Pten Lkb1-deficient endometrial tumors (lanes 1–4) or mouse tail (lanes 5) were extracted. PCR analysis was done as previously described (1, 2). Supplementary Figure 3. Representative images of immunohistochemical staining of Pten (A) and Lkb1 (B) proteins in Pten Lkb1-deficient endometrial tumors counterstained with hematoxylin. Bars, 25μM. Normal uterus was shown as a control. Supplementary Figure 4. Representative histopathology images of normal uterus (A) and precursor endometrial lesions (B) from Pten loxp/loxp Lkb1 loxp/loxp female mice with endometrium-specific co-deletion of Pten and Lkb1 (2 weeks post intrauterine injection of Ade-Cre). Scale bars, 25μM. Supplementary Figure 5. Representative images of histopathology of endometrial lesions from Pten loxp/loxp female mice with endometrium-specific deletion of Pten. Mice were sacrificed 10 months post intrauterine injection of Ade-Cre. Scale bars, 50μM. Supplementary Figure 6. Representative histopathology images of endometrial lesions from Lkb1loxp/loxp female mice with endometrium-specific deletion of Lkb1. Mice were sacrificed 10 months post intrauterine injection of Ade-Cre. Scale bars, 100μM. Supplementary Figure 7. Representative images of immunohistochemical staining of CK8 in Pten Lkb1-deficient endometrial tumors. Scale bars, 50μM (left panel); 25μM (right panel). Supplementary Figure 8. Representative histopathology images of metastases found in the lungs of Ptenloxp/loxp Lkb1loxp/loxp female mice bearing Pten Lkb1-deficient endometrial tumors. Bars, 50μM. Supplementary Figure 9. Representative histopathology image of Pten Lkb1-deficient endometrial cancer lesions with desmoplastic stromal response. Scale bar, 25μM. Supplementary Figure 10. Representative images of immunohistochemical staining of proteins as indicated in normal uterus (upper panels, A and B) and Pten Lkb1-deficient endometrial tumors (lower panels, A and B). Scale bars, 25μM.Supplementary Figure 11. Western blot analysis of LKB1 expression in ETN-1 and HEC108 cells with stable expression of vector or flag-tagged LKB1 wild-type. Vinculin was used a loading control.
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ARTICLE ABSTRACT
Signals from the tumor suppressors PTEN and LKB1 converge on mTOR to negatively regulate its function in cancer cells. Notably, both of these suppressors are attenuated in a significant fraction of human endometrial tumors. In this study, we generated a genetic mouse model of endometrial cancer driven by concomitant loss of these suppressors to gain pathophysiological insight into this disease. Dual loss of Pten and Lkb1 in the endometrial epithelium led to rapid development of advanced endometrioid endometrial tumors with 100% penetrance and short host survival. The tumors displayed dysregulated phosphatidylinositol 3-kinase (PI3K)/Akt and Lkb1/Ampk signaling with hyperactivation of mTOR signaling. Treatment with a dual PI3K/mTOR inhibitor, BEZ235, extended the time before tumor onset and prolonged overall survival. The PI3K inhibitor GDC-0941 used as a single agent reduced the growth rate of primary tumor implants in Pten/Lkb1-deficient mice, and the mTOR inhibitor RAD001 was unexpectedly as effective as BEZ235 in triggering tumor regression. In parallel, we also found that ectopic expression of LKB1 in PTEN/LKB1-deficient human endometrial cancer cells increased their sensitivity to PI3K inhibition. Together, our results demonstrated that Pten/Lkb1-deficient endometrial tumors rely strongly on deregulated mTOR signaling, and they provided evidence that LKB1 status may modulate the response of PTEN-deficient tumors to PI3K or mTOR inhibitors. Cancer Res; 74(1); 15–23. ©2013 AACR.Usage metrics
