The acquisition of an invasive phenotype by epithelial cells occurs through a loss of cellular ad... more The acquisition of an invasive phenotype by epithelial cells occurs through a loss of cellular adhesion and polarity, heralding a multistep process that leads to metastatic dissemination. Since its characterization in 1995, epithelial-mesenchymal transition (EMT) has been closely linked to the metastatic process. As a defining aspect of EMT, loss of cell adhesion through downregulation of E-cadherin is carried out by several transcriptional repressors; key among them the SNAI family of transcription factors. Here we identify for the first time that Lyn kinase functions as a key modulator of SNAI family protein localization and stability through control of the Vav-Rac1-PAK1 (Vav-Rac1-p21-activated kinase) pathway. Accordingly, targeting Lyn in vitro reduces EMT and in vivo reduces metastasis of primary tumors. We also demonstrate the clinical relevance of targeting Lyn as a key player controlling EMT; patient samples across many cancers revealed a strong negative correlation between Lyn and E-cadherin, and high Lyn expression in metastatic tumors as well as metastasis-prone primary tumors. This work reveals a novel pancancer mechanism of Lyndependent control of EMT and further underscores the role of this kinase in tumor progression.
Supplementary Figure S1. GSEA analyses of subtype-specific genes (PCS1, PCS2 and PCS3) in 42D ENZ... more Supplementary Figure S1. GSEA analyses of subtype-specific genes (PCS1, PCS2 and PCS3) in 42D ENZR and 42F ENZR cells. Supplementary Figure S2. A) Expression of FOXM1 in different cell lines. Total proteins were extracted from LNCaP, 16D CRPC and ENZ R 42D and 42F cells and western blot was performed using FOXM1 antibody (Abcam, 1:1000). Vinculin (1:5000) was used as a loading control. FOXM1 band densitometry was normalized to vinculin. B) Upstream regulator analysis of FOXM1 in PCS1 signature and 42D ENZR and 42F ENZR cells assessed by IPA (red: target gene overexpression, green: target gene downregulation, orange arrow: target gene is known as induced and blue arrow: target gene known as reduced by pathway activation). C) Expression of FOXM1 pathway expression in metastatic prostate cancer patients analysed using Genesapiens database (20). Supplementary Figure S3. A) Effect of Mon and Thiostrepton effect on FOXM1 transcriptional activity, 42D ENZR cells were treated with 100 nM of Mon and Thiostrepton for 24 hours and FOXM1 luciferase activity was performed. B) Monensin reduces FOXM1 protein level. Monensin effect (0, 1, 10, 50 and 100 nM) on FOXM1 protein was addressed using Western blot in 42D ENZR cells. Total protein were extracted and western blot was performed using FOXM1 antibody (1:1000). Vinculin (1:5000) was used as a loading control. C) Effect of Mon on cell cycle. Flow cytometry was performed to analyse cell cycle population after Mon treatment for 0 to 72h, percentage of sub-G1 cells in response to Mon in 16D CRPC , 42D ENZR and 42F ENZR cells was evaluated. D) DARTS assay in presence and absence of Mon in pronase treated samples indicated Mon binding to FOXM1. GAPDH was used as a control. E) Analysis of the effect of Mon on FOXM1 pathway by IPA Upstream regulator analysis of Mon exposed 42D ENZR cells (green: target gene downregulation, blue arrow: target gene known as reduced by pathway inactivation). Supplementary Figure S4. Portion of high CD49b (CD49b + /CD24-) cells in 16D ENZR , 42D ENZR and 42F ENZR cells. B) Effect of Mon and C) FOXM1 silencing by FOXM1 siRNA on portion of CD49b + /CD24cells. Supplementary Figure S5. The effect of FOXM1 inhibition on ALDH activity. A) Portion of ALDH active cells in 16D ENZR , 42D ENZR and 42F ENZR cells. B) The effect of Mon and C) FOXM1 silencing on ALDH activity in 42D ENZR and 42F ENZR cells. D) Cell proliferation of ALDH High and ALDH Low cells (separated from 42D ENZR and 42F ENZR cells using FACS and Aldefluor ALDH assay) in response to Mon measured by CTG cell proliferation assay. E) FOXM1 protein expression in
Supplementary Table S3. Up and downregulated genes in response to Mon in 42DENZR cells assessed b... more Supplementary Table S3. Up and downregulated genes in response to Mon in 42DENZR cells assessed by gene expression profiling.
Supplementary Figure S1. Generation of ENZR Xenografts and Cell Lines. Supplementary Figure S2. A... more Supplementary Figure S1. Generation of ENZR Xenografts and Cell Lines. Supplementary Figure S2. AR non-driven ENZR cells display a NE differentiation signature. Supplementary Figure S3. BRN2 expression is associated with PCa progression and a NE phenotype in human tumors. Supplementary Figure S4. BRN2 expression is inversely related to AR activity and is required for ENZ-induced NE marker expression in CRPC. Supplementary Figure S5. BRN2 overexpression induces NE marker expression and reciprocally regulates AR. Supplementary Figure S6. BRN2 activity and markers of NE differentiation are suppressed by androgens in CRPC and ENZR cells. Supplementary Figure S7. SOX2 expression is induced by ENZ and suppressed by androgens. Supplementary Figure S8. BRN2 is required for neuroendocrine marker expression and aggressive growth of CRPC cells in vitro.
Extracellular signal-regulated kinase (ERK)/mitogen-activated protein kinase pathway activity is ... more Extracellular signal-regulated kinase (ERK)/mitogen-activated protein kinase pathway activity is regulated by the antagonist function of activating kinases and inactivating protein phosphatases. Sustained ERK pathway activity is commonly observed in human malignancies; however, the mechanisms by which the pathway is protected from phosphatase-mediated inactivation in the tumor tissue remain obscure. Here, we show that methylesterase PME-1–mediated inhibition of the protein phosphatase 2A promotes basal ERK pathway activity and is required for efficient growth factor response. Mechanistically, PME-1 is shown to support ERK pathway signaling upstream of Raf, but downstream of growth factor receptors and protein kinase C. In malignant gliomas, PME-1 expression levels correlate with both ERK activity and cell proliferation in vivo. Moreover, PME-1 expression significantly correlates with disease progression in human astrocytic gliomas (n = 222). Together, these observations identify PME-1 expression as one mechanism by which ERK pathway activity is maintained in cancer cells and suggest an important functional role for PME-1 in the disease progression of human astrocytic gliomas. [Cancer Res 2009;69(7):2870–7]
Background: Androgen ablation remains the most effective therapy for patients with advanced disea... more Background: Androgen ablation remains the most effective therapy for patients with advanced disease. Unfortunately, most patients progress to castrate resistant prostate cancer (CRPC) characterized with hyper-activation of the androgen receptor (AR). Enzalutamide, a potent AR inhibitor showed efficacy by prolonging survival in CRPC patients. However, ENZ resistance (ENZR) rapidly occurs in patients and in our pre-clinical model targeting AR in ENZ resistant tumors with a 3rd generation AR inhibitor is short lived. The role of signal transducer and activator of transcription (STAT) 3 in the progression of prostate cancer is well established and has been repeatedly linked with maintenance of a stem cell phenotype across cancers. Additionally, drug resistance has been hypothesized to occur via enrichment of cancer stem-like cells (CSC), a phenotype associated with poor survival in patients. Therefore, we propose a shift of focus to target CSC phenotype using small molecule inhibitor of STAT3 called GPA500, instead of the AR axis to deal with ENZ resistance. Methods and Results: We developed a unique model of ENZ-resistance and found that cell lines derived from serially passaged ENZR tumors displayed broad genetic diversity and differential AR activity. Notably, the cell lines 42DENZR and 42FENZR are PSAlow, harbor an expanded CSC population and have STAT3 hyper-activation compared to CRPC controls measured by STAT3-luc reporter. Accordingly, Crystal Violet and MTT proliferation assays showed that 42DENZR and 42FENZR cell lines were more sensitive to the STAT3 inhibitor GPA500 compared to CRPC control. Targeting the ENZ-R cells with GPA500 reduced mRNA levels of CD133, CD44, SOX2, OCT4 and Nanog. The reduction of these CSC markers was accompanied by reduced self-renewal capacity measured by spheroid assay. Cytometry analysis revealed that treatment with GPA500 reduced α2β1+, CD44+ and CD133+ (CSC) population in the 42D and 42F cells. Conclusion: In this study, we provide pre-clinical proof that targeting the STAT3 pathway using GPA500 in ENZ resistance as well as CRPC reduces cell proliferation as well as expression of stem cell markers. Impact: Exploring mechanisms of ENZR resistance serves a critical unmet need in PCa oncology, as the number of men with ENZ resistant CRPC continues to rise. Targeting STAT3 with the small molecule inhibitor GPA500 may provide an effective method to treat ENZR patients or delay the emergence of ENZ resistance in CRPC by reducing the emergence of stem cells. Citation Format: Sepideh Vahid, Daksh Thaper, Alistair Davies, Micaela Janse Van Rensburg, Kate Gibson, Krisi Ketola, Martin Johansson, Jennifer L. Bishop, Amina Zoubeidi. Galiellalactone derivative targets stem cell population in ENZ-resistant prostate cancer through inhibition of STAT3. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3342.
Introduction: Metastasis is the most common cause of death from cancer and occurs when malignant ... more Introduction: Metastasis is the most common cause of death from cancer and occurs when malignant cells discard epithelial restraints and acquire invasive abilities, facilitating their dissemination to permissive micro-environments. This process is enhanced by tumor cell activation of Epithelial Mesenchymal Transition (EMT), a (normally embryonic) developmental program in which epithelial cells assume a mesenchymal phenotype during gastrulation and organogenesis, allowing single cell invasive movement away from the ectodermal layer. Recent evidence strongly implicates EMT induction in malignant progression and treatment resistance. For example, EMT regulatory transcription factors are required for breast cancer metastasis. Several oncogenic pathways (growth factors, Src family, MAPK, AKT) induce EMT. Lyn tyrosine kinase, a member of Src family tyrosine kinase is up-regulated in advanced prostate cancer and has been reported to correlate with aggressive breast cancer. Our objective is to determine the role of Lyn tyrosine kinase in EMT. Methods: LNCaP (Lymph Node Metastasis of Prostate Cancer), BT-549 (Triple Negative Breast Cancer) and UM-UC-13 (Bladder Cancer) cells were transfected with Lyn siRNA; EMT markers were monitored by western blot and qRT-PCR and immunofluorescence, migration by scratch assay and invasion by Boyden chamber. Sub cellular localization of proteins was examined by IF and nuclear/cyto extraction. In vivo experiments were performed in UC13-luc cells with shRNA of Lyn. Results: Here we report that Lyn expression is low in epithelial cells and is up-regulated in mesenchymal cells. Targeting Lyn using siRNA decreases EMT markers (Fibronectin, Vimentin and Zeb-1) at both mRNA and protein levels while increasing the epithelial marker (E-cadherin). Moreover, we found that Lyn siRNA decreases cell migration and invasion. Thisis decrease in mesenchymal phenotype can be attributed to the decrease in the amount of Slug and Snail, transcriptional repressors of E-Cadherin and activator of Vimentin and Fibronectin. Interestingly, we found that Lyn knockout induces a decrease of SLUG only at protein levels and not at mRNA levels. We discovered that Lyn triggers a signaling cascade through Vav-Rac-Pak1 pathway to alter sub cellular localization of the SNAI proteins leading to their proteasomal degradation. This effect results in decreased invasion and migration in vitro as well as decreased metastasis in vivo. Conclusion: Expression of Lyn kinase can be correlated to low prognosis and aggressive/metastatic phenotype. We show that knocking down Lyn by siRNA initiates a switch to a more epithelial phenotype reducing cell migration and invasion. Citation Format: Daksh Thaper, Sepideh Vahid, Ka Mun Nip, Igor Moskalev, Sebastian Frees, Morgan E. Roberts, Krisi Ketola, Kenneth W. Harder, Jennifer L. Bishop, Amina Zoubeidi. Lyn drives cancer metastasis via post-translational regulation of SNAI proteins. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1682.
Supplementary Figure S1. Generation of ENZR Xenografts and Cell Lines. Supplementary Figure S2. A... more Supplementary Figure S1. Generation of ENZR Xenografts and Cell Lines. Supplementary Figure S2. AR non-driven ENZR cells display a NE differentiation signature. Supplementary Figure S3. BRN2 expression is associated with PCa progression and a NE phenotype in human tumors. Supplementary Figure S4. BRN2 expression is inversely related to AR activity and is required for ENZ-induced NE marker expression in CRPC. Supplementary Figure S5. BRN2 overexpression induces NE marker expression and reciprocally regulates AR. Supplementary Figure S6. BRN2 activity and markers of NE differentiation are suppressed by androgens in CRPC and ENZR cells. Supplementary Figure S7. SOX2 expression is induced by ENZ and suppressed by androgens. Supplementary Figure S8. BRN2 is required for neuroendocrine marker expression and aggressive growth of CRPC cells in vitro.
The acquisition of an invasive phenotype by epithelial cells occurs through a loss of cellular ad... more The acquisition of an invasive phenotype by epithelial cells occurs through a loss of cellular adhesion and polarity, heralding a multistep process that leads to metastatic dissemination. Since its characterization in 1995, epithelial-mesenchymal transition (EMT) has been closely linked to the metastatic process. As a defining aspect of EMT, loss of cell adhesion through downregulation of E-cadherin is carried out by several transcriptional repressors; key among them the SNAI family of transcription factors. Here we identify for the first time that Lyn kinase functions as a key modulator of SNAI family protein localization and stability through control of the Vav-Rac1-PAK1 (Vav-Rac1-p21-activated kinase) pathway. Accordingly, targeting Lyn in vitro reduces EMT and in vivo reduces metastasis of primary tumors. We also demonstrate the clinical relevance of targeting Lyn as a key player controlling EMT; patient samples across many cancers revealed a strong negative correlation between Lyn and E-cadherin, and high Lyn expression in metastatic tumors as well as metastasis-prone primary tumors. This work reveals a novel pancancer mechanism of Lyndependent control of EMT and further underscores the role of this kinase in tumor progression.
Supplementary Figure S1. GSEA analyses of subtype-specific genes (PCS1, PCS2 and PCS3) in 42D ENZ... more Supplementary Figure S1. GSEA analyses of subtype-specific genes (PCS1, PCS2 and PCS3) in 42D ENZR and 42F ENZR cells. Supplementary Figure S2. A) Expression of FOXM1 in different cell lines. Total proteins were extracted from LNCaP, 16D CRPC and ENZ R 42D and 42F cells and western blot was performed using FOXM1 antibody (Abcam, 1:1000). Vinculin (1:5000) was used as a loading control. FOXM1 band densitometry was normalized to vinculin. B) Upstream regulator analysis of FOXM1 in PCS1 signature and 42D ENZR and 42F ENZR cells assessed by IPA (red: target gene overexpression, green: target gene downregulation, orange arrow: target gene is known as induced and blue arrow: target gene known as reduced by pathway activation). C) Expression of FOXM1 pathway expression in metastatic prostate cancer patients analysed using Genesapiens database (20). Supplementary Figure S3. A) Effect of Mon and Thiostrepton effect on FOXM1 transcriptional activity, 42D ENZR cells were treated with 100 nM of Mon and Thiostrepton for 24 hours and FOXM1 luciferase activity was performed. B) Monensin reduces FOXM1 protein level. Monensin effect (0, 1, 10, 50 and 100 nM) on FOXM1 protein was addressed using Western blot in 42D ENZR cells. Total protein were extracted and western blot was performed using FOXM1 antibody (1:1000). Vinculin (1:5000) was used as a loading control. C) Effect of Mon on cell cycle. Flow cytometry was performed to analyse cell cycle population after Mon treatment for 0 to 72h, percentage of sub-G1 cells in response to Mon in 16D CRPC , 42D ENZR and 42F ENZR cells was evaluated. D) DARTS assay in presence and absence of Mon in pronase treated samples indicated Mon binding to FOXM1. GAPDH was used as a control. E) Analysis of the effect of Mon on FOXM1 pathway by IPA Upstream regulator analysis of Mon exposed 42D ENZR cells (green: target gene downregulation, blue arrow: target gene known as reduced by pathway inactivation). Supplementary Figure S4. Portion of high CD49b (CD49b + /CD24-) cells in 16D ENZR , 42D ENZR and 42F ENZR cells. B) Effect of Mon and C) FOXM1 silencing by FOXM1 siRNA on portion of CD49b + /CD24cells. Supplementary Figure S5. The effect of FOXM1 inhibition on ALDH activity. A) Portion of ALDH active cells in 16D ENZR , 42D ENZR and 42F ENZR cells. B) The effect of Mon and C) FOXM1 silencing on ALDH activity in 42D ENZR and 42F ENZR cells. D) Cell proliferation of ALDH High and ALDH Low cells (separated from 42D ENZR and 42F ENZR cells using FACS and Aldefluor ALDH assay) in response to Mon measured by CTG cell proliferation assay. E) FOXM1 protein expression in
Supplementary Table S3. Up and downregulated genes in response to Mon in 42DENZR cells assessed b... more Supplementary Table S3. Up and downregulated genes in response to Mon in 42DENZR cells assessed by gene expression profiling.
Supplementary Figure S1. Generation of ENZR Xenografts and Cell Lines. Supplementary Figure S2. A... more Supplementary Figure S1. Generation of ENZR Xenografts and Cell Lines. Supplementary Figure S2. AR non-driven ENZR cells display a NE differentiation signature. Supplementary Figure S3. BRN2 expression is associated with PCa progression and a NE phenotype in human tumors. Supplementary Figure S4. BRN2 expression is inversely related to AR activity and is required for ENZ-induced NE marker expression in CRPC. Supplementary Figure S5. BRN2 overexpression induces NE marker expression and reciprocally regulates AR. Supplementary Figure S6. BRN2 activity and markers of NE differentiation are suppressed by androgens in CRPC and ENZR cells. Supplementary Figure S7. SOX2 expression is induced by ENZ and suppressed by androgens. Supplementary Figure S8. BRN2 is required for neuroendocrine marker expression and aggressive growth of CRPC cells in vitro.
Extracellular signal-regulated kinase (ERK)/mitogen-activated protein kinase pathway activity is ... more Extracellular signal-regulated kinase (ERK)/mitogen-activated protein kinase pathway activity is regulated by the antagonist function of activating kinases and inactivating protein phosphatases. Sustained ERK pathway activity is commonly observed in human malignancies; however, the mechanisms by which the pathway is protected from phosphatase-mediated inactivation in the tumor tissue remain obscure. Here, we show that methylesterase PME-1–mediated inhibition of the protein phosphatase 2A promotes basal ERK pathway activity and is required for efficient growth factor response. Mechanistically, PME-1 is shown to support ERK pathway signaling upstream of Raf, but downstream of growth factor receptors and protein kinase C. In malignant gliomas, PME-1 expression levels correlate with both ERK activity and cell proliferation in vivo. Moreover, PME-1 expression significantly correlates with disease progression in human astrocytic gliomas (n = 222). Together, these observations identify PME-1 expression as one mechanism by which ERK pathway activity is maintained in cancer cells and suggest an important functional role for PME-1 in the disease progression of human astrocytic gliomas. [Cancer Res 2009;69(7):2870–7]
Background: Androgen ablation remains the most effective therapy for patients with advanced disea... more Background: Androgen ablation remains the most effective therapy for patients with advanced disease. Unfortunately, most patients progress to castrate resistant prostate cancer (CRPC) characterized with hyper-activation of the androgen receptor (AR). Enzalutamide, a potent AR inhibitor showed efficacy by prolonging survival in CRPC patients. However, ENZ resistance (ENZR) rapidly occurs in patients and in our pre-clinical model targeting AR in ENZ resistant tumors with a 3rd generation AR inhibitor is short lived. The role of signal transducer and activator of transcription (STAT) 3 in the progression of prostate cancer is well established and has been repeatedly linked with maintenance of a stem cell phenotype across cancers. Additionally, drug resistance has been hypothesized to occur via enrichment of cancer stem-like cells (CSC), a phenotype associated with poor survival in patients. Therefore, we propose a shift of focus to target CSC phenotype using small molecule inhibitor of STAT3 called GPA500, instead of the AR axis to deal with ENZ resistance. Methods and Results: We developed a unique model of ENZ-resistance and found that cell lines derived from serially passaged ENZR tumors displayed broad genetic diversity and differential AR activity. Notably, the cell lines 42DENZR and 42FENZR are PSAlow, harbor an expanded CSC population and have STAT3 hyper-activation compared to CRPC controls measured by STAT3-luc reporter. Accordingly, Crystal Violet and MTT proliferation assays showed that 42DENZR and 42FENZR cell lines were more sensitive to the STAT3 inhibitor GPA500 compared to CRPC control. Targeting the ENZ-R cells with GPA500 reduced mRNA levels of CD133, CD44, SOX2, OCT4 and Nanog. The reduction of these CSC markers was accompanied by reduced self-renewal capacity measured by spheroid assay. Cytometry analysis revealed that treatment with GPA500 reduced α2β1+, CD44+ and CD133+ (CSC) population in the 42D and 42F cells. Conclusion: In this study, we provide pre-clinical proof that targeting the STAT3 pathway using GPA500 in ENZ resistance as well as CRPC reduces cell proliferation as well as expression of stem cell markers. Impact: Exploring mechanisms of ENZR resistance serves a critical unmet need in PCa oncology, as the number of men with ENZ resistant CRPC continues to rise. Targeting STAT3 with the small molecule inhibitor GPA500 may provide an effective method to treat ENZR patients or delay the emergence of ENZ resistance in CRPC by reducing the emergence of stem cells. Citation Format: Sepideh Vahid, Daksh Thaper, Alistair Davies, Micaela Janse Van Rensburg, Kate Gibson, Krisi Ketola, Martin Johansson, Jennifer L. Bishop, Amina Zoubeidi. Galiellalactone derivative targets stem cell population in ENZ-resistant prostate cancer through inhibition of STAT3. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3342.
Introduction: Metastasis is the most common cause of death from cancer and occurs when malignant ... more Introduction: Metastasis is the most common cause of death from cancer and occurs when malignant cells discard epithelial restraints and acquire invasive abilities, facilitating their dissemination to permissive micro-environments. This process is enhanced by tumor cell activation of Epithelial Mesenchymal Transition (EMT), a (normally embryonic) developmental program in which epithelial cells assume a mesenchymal phenotype during gastrulation and organogenesis, allowing single cell invasive movement away from the ectodermal layer. Recent evidence strongly implicates EMT induction in malignant progression and treatment resistance. For example, EMT regulatory transcription factors are required for breast cancer metastasis. Several oncogenic pathways (growth factors, Src family, MAPK, AKT) induce EMT. Lyn tyrosine kinase, a member of Src family tyrosine kinase is up-regulated in advanced prostate cancer and has been reported to correlate with aggressive breast cancer. Our objective is to determine the role of Lyn tyrosine kinase in EMT. Methods: LNCaP (Lymph Node Metastasis of Prostate Cancer), BT-549 (Triple Negative Breast Cancer) and UM-UC-13 (Bladder Cancer) cells were transfected with Lyn siRNA; EMT markers were monitored by western blot and qRT-PCR and immunofluorescence, migration by scratch assay and invasion by Boyden chamber. Sub cellular localization of proteins was examined by IF and nuclear/cyto extraction. In vivo experiments were performed in UC13-luc cells with shRNA of Lyn. Results: Here we report that Lyn expression is low in epithelial cells and is up-regulated in mesenchymal cells. Targeting Lyn using siRNA decreases EMT markers (Fibronectin, Vimentin and Zeb-1) at both mRNA and protein levels while increasing the epithelial marker (E-cadherin). Moreover, we found that Lyn siRNA decreases cell migration and invasion. Thisis decrease in mesenchymal phenotype can be attributed to the decrease in the amount of Slug and Snail, transcriptional repressors of E-Cadherin and activator of Vimentin and Fibronectin. Interestingly, we found that Lyn knockout induces a decrease of SLUG only at protein levels and not at mRNA levels. We discovered that Lyn triggers a signaling cascade through Vav-Rac-Pak1 pathway to alter sub cellular localization of the SNAI proteins leading to their proteasomal degradation. This effect results in decreased invasion and migration in vitro as well as decreased metastasis in vivo. Conclusion: Expression of Lyn kinase can be correlated to low prognosis and aggressive/metastatic phenotype. We show that knocking down Lyn by siRNA initiates a switch to a more epithelial phenotype reducing cell migration and invasion. Citation Format: Daksh Thaper, Sepideh Vahid, Ka Mun Nip, Igor Moskalev, Sebastian Frees, Morgan E. Roberts, Krisi Ketola, Kenneth W. Harder, Jennifer L. Bishop, Amina Zoubeidi. Lyn drives cancer metastasis via post-translational regulation of SNAI proteins. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1682.
Supplementary Figure S1. Generation of ENZR Xenografts and Cell Lines. Supplementary Figure S2. A... more Supplementary Figure S1. Generation of ENZR Xenografts and Cell Lines. Supplementary Figure S2. AR non-driven ENZR cells display a NE differentiation signature. Supplementary Figure S3. BRN2 expression is associated with PCa progression and a NE phenotype in human tumors. Supplementary Figure S4. BRN2 expression is inversely related to AR activity and is required for ENZ-induced NE marker expression in CRPC. Supplementary Figure S5. BRN2 overexpression induces NE marker expression and reciprocally regulates AR. Supplementary Figure S6. BRN2 activity and markers of NE differentiation are suppressed by androgens in CRPC and ENZR cells. Supplementary Figure S7. SOX2 expression is induced by ENZ and suppressed by androgens. Supplementary Figure S8. BRN2 is required for neuroendocrine marker expression and aggressive growth of CRPC cells in vitro.
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Papers by Kirsi Ketola