The Experts below are selected from a list of 360 Experts worldwide ranked by ideXlab platform
Peter E. Lobie - One of the best experts on this subject based on the ideXlab platform.
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trefoil factor 3 tff3 stimulates de novo angiogenesis in mammary carcinoma both directly and indirectly via il 8 cxcr2
PLOS ONE, 2015Co-Authors: Vijay Pandey, Xiangjun Kong, Zhengsheng Wu, Xiaonan Wang, Peter E. LobieAbstract:Mammary carcinoma cells produce pro-angiogenic factors to stimulate angiogenesis and tumor growth. Trefoil factor-3 (TFF3) is an oncogene secreted from mammary carcinoma cells and associated with poor prognosis. Herein, we demonstrate that TFF3 produced in mammary carcinoma cells functions as a promoter of tumor angiogenesis. Forced Expression of TFF3 in mammary carcinoma cells promoted proliferation, survival, invasion and in vitro tubule formation of human umbilical vein endothelial cells (HUVEC). MCF7-TFF3 cells with Forced Expression of TFF3 generated tumors with enhanced microvessel density as compared to tumors formed by vector control cells. Depletion of TFF3 in mammary carcinoma cells by siRNA concordantly decreased the angiogenic behavior of HUVEC. Forced Expression of TFF3 in mammary carcinoma cells stimulated IL-8 transcription and subsequently enhanced IL-8 Expression in both mammary carcinoma cells and HUVEC. Depletion of IL-8 in mammary carcinoma cells with Forced Expression of TFF3, or antibody inhibition of IL-8, partially abrogated mammary carcinoma cell TFF3-stimulated HUVEC angiogenic behavior in vitro, as did inhibition of the IL-8 receptor, CXCR2. Depletion of STAT3 by siRNA in MCF-7 cells with Forced Expression of TFF3 partially diminished the angiogenic capability of TFF3 on stimulation of cellular processes of HUVEC. Exogenous recombinant hTFF3 also directly promoted the angiogenic behavior of HUVEC. Hence, TFF3 is a potent angiogenic factor and functions as a promoter of de novo angiogenesis in mammary carcinoma, which may co-coordinate with the growth promoting and metastatic actions of TFF3 in mammary carcinoma to enhance tumor progression.
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trefoil factor 3 promotes metastatic seeding and predicts poor survival outcome of patients with mammary carcinoma
Breast Cancer Research, 2014Co-Authors: Vijay Pandey, Zhengsheng Wu, Min Zhang, Rui Li, Jian Zhang, Peter E. LobieAbstract:Introduction: Recurrence or early metastasis remains the predominant cause of mortality in patients with estrogen receptor positive (ER+) mammary carcinoma (MC). However, the molecular mechanisms underlying the initial progression of ER+ MC to metastasis remains poorly understood. Trefoil factor 3 (TFF3) is an estrogen-responsive oncogene in MC. Herein, we provide evidence for a functional role of TFF3 in metastatic progression of ER+ MC. Methods: The association of TFF3 Expression with clinicopathological parameters and survival outcome in a cohort of MC patients was assessed by immunohistochemistry. The Expression of TFF3 in MCF7 and T47D cells was modulated by Forced Expression or siRNA-mediated depletion of TFF3. mRNA and protein levels were determined using qPCR and western blot. The functional effect of modulation of TFF3 Expression in MC cells was determined in vitro and in vivo. Mechanistic analyses were performed using reporter constructs, modulation of signal transducer and activator of transcription 3 (STAT3) Expression, and pharmacological inhibitors against c-SRC and STAT3 activity. Results: TFF3 protein Expression was positively associated with larger tumour size, lymph node metastasis, higher stage, and poor survival outcome. Forced Expression of TFF3 in ER+ MC cells stimulated colony scattering, cell adhesion to a Collagen I-coated matrix, colony formation on a Collagen I- or Matrigel-coated matrix, endothelial cell adhesion, and transmigration through an endothelial cell barrier. In vivo, Forced Expression of TFF3 in MCF7 cells stimulated the formation of metastatic nodules in animal lungs. TFF3 regulation of the mRNA levels of epithelial, mesenchymal, and metastatic-related genes in ER+ MC cells were consistent with the altered cell behaviour. Forced Expression of TFF3 in ER+ MC cells stimulated phosphorylation of c-SRC that subsequently increased STAT3 activity, which lead to the downregulation of E-cadherin. siRNA-mediated depletion of TFF3 reduced the invasiveness of ER+ MC cells. Conclusions: TFF3 Expression predicts metastasis and poor survival outcome of patients with MC and functionally stimulates cellular invasion and metastasis of ER+ MC cells. Adjuvant functional inhibition of TFF3 may therefore be considered to ameliorate outcome of ER+ MC patients.
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p4 02 05 the regulation of artemin signalling by igf 1 in mammary carcinoma cells
Cancer Research, 2011Co-Authors: C Yip, Peter E. Lobie, Dong-xu Liu, Jk PerryAbstract:Artemin is a neurotrophic signalling factor which belongs to the glial-derived neurotrophic factor (GDNF) family of ligands. Artemin acts as a survival, proliferation and migration factor for a number of neurological cell types, by signalling through the RET (rearranged during transfection) receptor and, in most cases, the GDNF receptor (GFR)-a3 co-receptor. Recently, a number of published studies have implicated Artemin as a potential oncogene in several cell types, including mammary carcinoma cells. Other studies further indicate that Artemin may influence cancer progression and tamoxifen resistance in some breast cancers. Available clinical data has demonstrated that increased Artemin Expression is correlated with decreased overall patient survival in breast cancer patients and a poor outcome in tamoxifen treated breast cancer patients. Here we investigate interaction between the Artemin and the insulin-like growth factor-1 (IGF-1) signal transduction pathways. Using mammary carcinoma cell lines, we demonstrate that IGF-1 treatment increases the endogenous Expression of both Artemin and its endogenous receptors, RET and GFRa3. Semi-quantitative RT-PCR assays demonstrated that IGF-1 stimulated mRNA Expression of Artemin as well as RET and GFRa3 in wild-type MCF-7 and ZR-75-1 cells in a time-dependent and dose-dependent manner. The same effect was not observed in wild-type T47D cells where IGF-1 did not increase Artemin mRNA Expression. We also demonstrated that Forced Expression of Artemin in MCF-7 cells consistently enhanced the response of these cells to IGF-1 in a number of cell function assays. Forced Expression of Artemin significantly enhanced IGF-1-mediated stimulation of total cell number in MCF-7 cells. Consistent with this, Artemin enhanced IGF-1-mediated stimulation of S-phase entry and cell survival. In a soft agar assay, Forced Expression of Artemin also enhanced IGF-1-mediated stimulation of colony formation. Conversely, depletion of Artemin Expression using siRNA abrogated the response to IGF-1 stimulation in MCF-7 cells. Artemin depletion significantly decreased IGF-1-stimulated increase in total cell number by decreasing IGF-1-stimulated cell proliferation and protection from apoptotic cell death. In addition, Forced Expression of Artemin in MCF-7 cells reduced cell sensitivity to the IGF-1 receptor small molecule inhibitor, AG1024. In conclusion, we have demonstrated that IGF-1 increases Artemin mRNA and protein Expression in the breast cancer cell lines MCF-7 and ZR-75-1 and have identified potential cross-talk between the Artemin and IGF-1 signalling pathways in MCF-7 cells. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-02-05.
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artemin synergizes with twist1 to promote metastasis and poor survival outcome in patients with er negative mammary carcinoma
Breast Cancer Research, 2011Co-Authors: Arindam Banerjee, Peter E. Lobie, Jian Kang, Dong-xu Liu, Vijay Pandey, Tao Zhu, Pengxu QianAbstract:ARTEMIN (ARTN) is an estrogen regulated growth factor, the Expression of which promotes resistance to antiestrogen therapies and predicts poorer survival outcome of patients with estrogen receptor (ER) positive mammary carcinoma (ER+MC) treated with tamoxifen. ARTN is also expressed in ER negative mammary carcinoma (ER-MC). Herein, we determined the role of ARTN in ER-MC and defined the mechanism of action producing poor patient prognosis. We modulated the Expression of ARTN in two ER- (mesenchymal/claudin-low) mammary carcinoma cell lines (BT549 and MDA-MB-231) by Forced Expression or small interfering RNA (siRNA) mediated depletion. The effects of modulation of ARTN Expression were examined by various in vitro measures of oncogenicity, including the Expression of TWIST1 messenger RNA (mRNA) and protein. In vitro results were correlated to xenograft studies in immunodeficient mice. Co-Expression of ARTN and TWIST1 and their association to poor survival outcome were examined in a cohort of patients with ER-MC. Pathway analysis was performed by pharmacological inhibition of phosphorylation of AKT (pAKT-Ser 473) or modulation of TWIST1 Expression. ARTN Expression resulted in ER-MC cells with enhanced mesenchymal characteristics, including increased invasion and a gene Expression profile consistent with enhanced mesenchymal phenotype. ARTN stimulated ER-MC cell anchorage independent and 3D matrigel growth, endothelial cell adhesion and transmigration of ER-MC cells through an endothelial cell barrier. Forced Expression of ARTN produced a larger, locally invasive tumour mass with tumour emboli that produced distant metastasis. ARTN regulated TWIST1 Expression in ER-MC cells and ARTN Expression was significantly correlated to TWIST1 Expression in a panel of mammary carcinoma cell lines and in a cohort of patients with ER-MC. Low Expression of both ARTN and TWIST1 predicted 100% relapse free and overall survival in patients with ER-MC, whereas high Expression of both ARTN and TWIST1 was associated with a poor survival outcome. ARTN stimulated an increase in TWIST1 Expression via increased AKT activity. siRNA mediated depletion of TWIST1 abrogated ARTN stimulated cellular behaviour associated with metastasis, and Forced Expression of TWIST1 abrogated the functional effects of ARTN depletion. ARTN and TWIST1 synergize to produce a worse outcome in ER-MC and combined inhibition of ARTN and phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) may therefore provide a novel therapeutic strategy in this subtype of mammary carcinoma.
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trefoil factor 3 is oncogenic and mediates anti estrogen resistance in human mammary carcinoma
Neoplasia, 2010Co-Authors: Nagarajan Kannan, Kumarasamypet M. Mohankumar, Prudence M Grandison, Xiangjun Kong, Lance D Miller, Jianzhong Tang, Hichem C. Mertani, Jian Kang, Jo K Perry, Peter E. LobieAbstract:We report herein that trefoil factor 3 (TFF3) is oncogenic and mediates anti-estrogen resistance in human mammary carcinoma. Forced Expression of TFF3 in mammary carcinoma cells increased cell proliferation and survival, enhanced anchorage-independent growth, and promoted migration and invasion. Moreover, Forced Expression of TFF3 increased tumor size in xenograft models. Conversely, depletion of endogenous TFF3 with small interfering RNA (siRNA) decreased the oncogenicity and invasiveness of mammary carcinoma cells. Neutralization of secreted TFF3 by antibody promoted apoptosis, decreased cell growth in vitro, and arrested mammary carcinoma xenograft growth. TFF3 Expression was significantly correlated to decreased survival of estrogen receptor (ER)-positive breast cancer patients treated with tamoxifen. Forced Expression of TFF3 in mammary carcinoma cells increased ER transcriptional activity, promoted estrogen-independent growth, and produced resistance to tamoxifen and fulvestrant in vitro and to tamoxifen in xenograft models. siRNA-mediated depletion or antibody inhibition of TFF3 significantly enhanced the efficacy of antiestrogens. Increased TFF3 Expression was observed in tamoxifen-resistant (TAMR) cells and antibody inhibition of TFF3 in TAMR cells improved tamoxifen sensitivity. Functional antagonism of TFF3 therefore warrants consideration as a novel therapeutic strategy for mammary carcinoma.
Toshihiko Yanase - One of the best experts on this subject based on the ideXlab platform.
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adipose tissue derived and bone marrow derived mesenchymal cells develop into different lineage of steroidogenic cells by Forced Expression of steroidogenic factor 1
Endocrinology, 2008Co-Authors: Shigeki Gondo, Taijiro Okabe, Tomoko Tanaka, Hidetaka Morinaga, Masatoshi Nomura, Ryoichi Takayanagi, Hajime Nawata, Toshihiko YanaseAbstract:Steroidogenic factor 1 (SF-1)/adrenal 4 binding protein is an essential nuclear receptor for steroidogenesis, as well as for adrenal and gonadal gland development. We have previously clarified that adenovirus-mediated Forced Expression of SF-1 can transform long-term cultured mouse bone marrow mesenchymal cells (BMCs) into ACTH-responsive steroidogenic cells. In the present study, we extended this work to adipose tissue-derived mesenchymal cells (AMCs) and compared its steroidogenic capacity with those of BMCs prepared from the identical mouse. Several cell surface markers, including potential mesenchymal cell markers, were identical in both cell types, and, as expected, Forced Expression of SF-1 caused AMCs to be transformed into ACTH-responsive steroidogenic cells. However, more elaborate studies revealed that the steroidogenic property of AMCs was rather different from that of BMCs, especially in steroidogenic lineage. In response to increased SF-1 Expression and/or treatment with retinoic acid, AMCs w...
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adipose tissue derived and bone marrow derived mesenchymal cells develop into different lineage of steroidogenic cells by Forced Expression of steroidogenic factor 1
Endocrinology, 2008Co-Authors: Shigeki Gondo, Taijiro Okabe, Tomoko Tanaka, Hidetaka Morinaga, Masatoshi Nomura, Ryoichi Takayanagi, Hajime Nawata, Toshihiko YanaseAbstract:Steroidogenic factor 1 (SF-1)/adrenal 4 binding protein is an essential nuclear receptor for steroidogenesis, as well as for adrenal and gonadal gland development. We have previously clarified that adenovirus-mediated Forced Expression of SF-1 can transform long-term cultured mouse bone marrow mesenchymal cells (BMCs) into ACTH-responsive steroidogenic cells. In the present study, we extended this work to adipose tissue-derived mesenchymal cells (AMCs) and compared its steroidogenic capacity with those of BMCs prepared from the identical mouse. Several cell surface markers, including potential mesenchymal cell markers, were identical in both cell types, and, as expected, Forced Expression of SF-1 caused AMCs to be transformed into ACTH-responsive steroidogenic cells. However, more elaborate studies revealed that the steroidogenic property of AMCs was rather different from that of BMCs, especially in steroidogenic lineage. In response to increased SF-1 Expression and/or treatment with retinoic acid, AMCs were much more prone to produce adrenal steroid, corticosterone rather than gonadal steroid, testosterone, whereas the contrary was evident in BMCs. Such marked differences in steroidogenic profiles between AMCs and BMCs were also evident by the changes of steroidogenic enzymes. These novel results suggest a promising utility of AMCs for autologous cell regeneration therapy for patients with steroid insufficiency and also a necessity for appropriate tissue selection in preparing mesenchymal stem cells according to the aim. The different steroidogenic potency of AMCs or BMCs might provide a good model for the clarification of the mechanism of tissue- or cell-specific adrenal and gonadal steroidogenic cell differentiation.
Michael S Parmacek - One of the best experts on this subject based on the ideXlab platform.
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notch signaling represses myocardin induced smooth muscle cell differentiation
Journal of Biological Chemistry, 2005Co-Authors: Aaron Proweller, Warren S Pear, Michael S ParmacekAbstract:Abstract Notch signaling is essential for vascular patterning and response of the vasculature to injury and growth factor stimulation. Despite these findings, the molecular basis of Notch signaling in the vasculature is poorly understood. Here we report that activation of Notch signaling mediated through members of the HRT family of basic helix-loop-helix transcription factors represses smooth muscle cell (SMC) differentiation and Expression of genes encoding smooth muscle cell contractile markers. Activation of Notch receptors by Jagged1 or Forced Expression of the constitutively active Notch1 intracellular domain in C3H10T1/2 fibroblasts inhibited myocardin-dependent transcription of SMC-restricted genes and activity of multiple SMC-restricted transcriptional regulatory elements. Consistent with these findings, Forced Expression of HRT2 inhibited myocardin-induced Expression of SMC-restricted genes and activity of SMC-restricted transcriptional regulatory elements. Moreover, Forced Expression of HRT2 repressed transcription of multiple SMC-restricted transcriptional regulatory elements in A10 SMCs. The repressive function of HRT2 was not mediated via the capacity of HRT2 to bind SMC CArG elements or by disruption of myocardin-SRF protein complexes. Structure-function analyses of HRT2 indicated that repression required the basic DNA binding domain and additional C-terminal sequence. Taken together, these results demonstrate that Notch signaling represses myocardin-dependent SMC transcription. These data are consistent with a model wherein Notch signaling represses SMC differentiation and maintenance of the contractile SMC phenotype.
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gata 6 regulates genes promoting synthetic functions in vascular smooth muscle cells
Arteriosclerosis Thrombosis and Vascular Biology, 2004Co-Authors: John J Lepore, Thomas P Cappola, Patricia Mericko, Edward E Morrisey, Michael S ParmacekAbstract:Objective— Previous studies suggested the zinc-finger transcription factor GATA-6 inhibits vascular smooth muscle cell (VSMC) proliferation and promotes the contractile VSMC phenotype. The objective of this study was to identify bona fide target genes regulated by GATA-6 in VSMCs. Methods and Results— Microarray analyses were performed comparing mRNA from rat aortic smooth muscle cells (SMCs) infected with either adenovirus encoding a dominant-negative GATA-6/engrailed fusion protein or with control adenovirus. These studies identified 122 genes differentially expressed by at least 2-fold, including multiple genes involved in cell–cell signaling and cell–matrix interactions. Among these, endothelin-1 and the angiotensin type1a (AT1a) receptor are known to be induced in VSMCs in response to inflammatory stimuli and to be expressed in a GATA-dependent manner in cardiac myocytes in response to hemodynamic stress. Consistent with these findings, the endothelin-1 and AT1a receptor promoters were activated by Forced Expression of GATA-6 and repressed by Forced Expression of GATA-6/engrailed. Surprisingly, genes encoding SMC contractile proteins were not altered, and myocardin-induced SMC differentiation was not impaired in GATA-6−/− embryonic stem cells. Conclusions— These data demonstrate that in VSMCs, GATA-6 regulates a set of genes associated with synthetic SMC functions and suggest that this transcriptional pathway may be independent from myocardin-induced SMC differentiation.
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megakaryoblastic leukemia factor 1 transduces cytoskeletal signals and induces smooth muscle cell differentiation from undifferentiated embryonic stem cells
Journal of Biological Chemistry, 2004Co-Authors: Mary Chen, Patricia Mericko, John J Lepore, Michael S ParmacekAbstract:Abstract The SAP domain transcription factor myocardin plays a critical role in the transcriptional program regulating smooth muscle cell differentiation. In this report, we describe the capacity of myocardin to physically associate with megakaryoblastic leukemia factor-1 (MKL1) and characterize the function of MKL1 in smooth muscle cells (SMCs). The MKL1 gene is expressed in most human tissues and myocardin and MKL are co-expressed in SMCs. MKL1 and myocardin physically associate via conserved leucine zipper domains. OverExpression of MKL1 transactivates serum response factor (SRF)-dependent SMC-restricted transcriptional regulatory elements including the SM22α promoter, smooth muscle myosin heavy chain promoter/enhancer, and SM-α-actin promoter/enhancer in non-SMCs. Moreover, Forced Expression of MKL1 and SRF in undifferentiated SRF-/- embryonic stem cells activates multiple endogenous SMC-restricted genes at levels equivalent to, or exceeding, myocardin. Forced Expression of a dominant-negative MKL1 mutant reduces myocardin-induced activation of the SMC-specific SM22α promoter. In NIH3T3 fibroblasts MKL1 localizes to the cytoplasm and translocates to the nucleus in response to serum stimulation, actin treadmilling, and RhoA signaling. In contrast, in SMCs MKL1 is observed exclusively in the nucleus regardless of serum conditions or RhoA signaling. However, when actin polymerization is disrupted MKL1 translocates from the nucleus to the cytoplasm in SMCs. Together, these data were consistent with a model wherein MKL1 transduces signals from the cytoskeleton to the nucleus in SMCs and regulates SRF-dependent SMC differentiation autonomously or in concert with myocardin.
Shigeki Gondo - One of the best experts on this subject based on the ideXlab platform.
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adipose tissue derived and bone marrow derived mesenchymal cells develop into different lineage of steroidogenic cells by Forced Expression of steroidogenic factor 1
Endocrinology, 2008Co-Authors: Shigeki Gondo, Taijiro Okabe, Tomoko Tanaka, Hidetaka Morinaga, Masatoshi Nomura, Ryoichi Takayanagi, Hajime Nawata, Toshihiko YanaseAbstract:Steroidogenic factor 1 (SF-1)/adrenal 4 binding protein is an essential nuclear receptor for steroidogenesis, as well as for adrenal and gonadal gland development. We have previously clarified that adenovirus-mediated Forced Expression of SF-1 can transform long-term cultured mouse bone marrow mesenchymal cells (BMCs) into ACTH-responsive steroidogenic cells. In the present study, we extended this work to adipose tissue-derived mesenchymal cells (AMCs) and compared its steroidogenic capacity with those of BMCs prepared from the identical mouse. Several cell surface markers, including potential mesenchymal cell markers, were identical in both cell types, and, as expected, Forced Expression of SF-1 caused AMCs to be transformed into ACTH-responsive steroidogenic cells. However, more elaborate studies revealed that the steroidogenic property of AMCs was rather different from that of BMCs, especially in steroidogenic lineage. In response to increased SF-1 Expression and/or treatment with retinoic acid, AMCs w...
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adipose tissue derived and bone marrow derived mesenchymal cells develop into different lineage of steroidogenic cells by Forced Expression of steroidogenic factor 1
Endocrinology, 2008Co-Authors: Shigeki Gondo, Taijiro Okabe, Tomoko Tanaka, Hidetaka Morinaga, Masatoshi Nomura, Ryoichi Takayanagi, Hajime Nawata, Toshihiko YanaseAbstract:Steroidogenic factor 1 (SF-1)/adrenal 4 binding protein is an essential nuclear receptor for steroidogenesis, as well as for adrenal and gonadal gland development. We have previously clarified that adenovirus-mediated Forced Expression of SF-1 can transform long-term cultured mouse bone marrow mesenchymal cells (BMCs) into ACTH-responsive steroidogenic cells. In the present study, we extended this work to adipose tissue-derived mesenchymal cells (AMCs) and compared its steroidogenic capacity with those of BMCs prepared from the identical mouse. Several cell surface markers, including potential mesenchymal cell markers, were identical in both cell types, and, as expected, Forced Expression of SF-1 caused AMCs to be transformed into ACTH-responsive steroidogenic cells. However, more elaborate studies revealed that the steroidogenic property of AMCs was rather different from that of BMCs, especially in steroidogenic lineage. In response to increased SF-1 Expression and/or treatment with retinoic acid, AMCs were much more prone to produce adrenal steroid, corticosterone rather than gonadal steroid, testosterone, whereas the contrary was evident in BMCs. Such marked differences in steroidogenic profiles between AMCs and BMCs were also evident by the changes of steroidogenic enzymes. These novel results suggest a promising utility of AMCs for autologous cell regeneration therapy for patients with steroid insufficiency and also a necessity for appropriate tissue selection in preparing mesenchymal stem cells according to the aim. The different steroidogenic potency of AMCs or BMCs might provide a good model for the clarification of the mechanism of tissue- or cell-specific adrenal and gonadal steroidogenic cell differentiation.
Vijay Pandey - One of the best experts on this subject based on the ideXlab platform.
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Expression of two non-mutated genetic elements is sufficient to stimulate oncogenic transformation of human mammary epithelial cells
Nature Publishing Group, 2018Co-Authors: Vijay Pandey, Min Zhang, Mingliang You, Weijie Zhang, Rumei Chen, Wei Zhang, Tao ZhuAbstract:Abstract Trefoil factor 3 (TFF3) Expression is positively associated with advanced clinicopathological features of mammary carcinoma (MC). Herein, we provide evidence for a functional role of TFF3 in oncogenic transformation of immortalized, but otherwise normal human mammary epithelial cells (HMECs), namely, HMEC-hTERT, MCF10A, and MCF12A. Forced Expression of TFF3 in immortalized-HMECs enhanced cell proliferation, cell survival, anchorage-independent growth, produced highly disorganised three-dimensional (3D) acinar structures and generated tumours in immunocompromised mice. Forced Expression of TFF3 in immortalized-HMECs stimulated STAT3 activity that was required for TFF3-stimulated cell proliferation, survival, and anchorage-independent growth. TFF3 specifically utilised STAT3 activity to govern a transcriptional program, which was required for TFF3-stimulated oncogenic transformation of immortalized-HMECs, including transcriptional upregulation of CCND1 and BCL2. siRNA-mediated depletion or functional inhibition of STAT3 significantly inhibited the TFF3-stimulated transcription of CCND1 and BCL2 and oncogenicity in immortalized-HMECs. Furthermore, DOX-inducible Expression of TFF3 in HMEC-hTERT cells also permitted anchorage-independent growth and produced disorganized acinar structures in 3D Matrigel culture. Removal of DOX-induced Expression of TFF3 in HMEC-hTERT cells, previously grown with DOX, resulted in efficient normalisation of the disorganized acinar architecture and attenuated cell viability in Matrigel culture. Cumulatively, these findings suggest that TFF3 is a potent oncogene and its increased Expression along with hTERT in HMECs is sufficient to produce oncogenic transformation
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trefoil factor 3 tff3 stimulates de novo angiogenesis in mammary carcinoma both directly and indirectly via il 8 cxcr2
PLOS ONE, 2015Co-Authors: Vijay Pandey, Xiangjun Kong, Zhengsheng Wu, Xiaonan Wang, Peter E. LobieAbstract:Mammary carcinoma cells produce pro-angiogenic factors to stimulate angiogenesis and tumor growth. Trefoil factor-3 (TFF3) is an oncogene secreted from mammary carcinoma cells and associated with poor prognosis. Herein, we demonstrate that TFF3 produced in mammary carcinoma cells functions as a promoter of tumor angiogenesis. Forced Expression of TFF3 in mammary carcinoma cells promoted proliferation, survival, invasion and in vitro tubule formation of human umbilical vein endothelial cells (HUVEC). MCF7-TFF3 cells with Forced Expression of TFF3 generated tumors with enhanced microvessel density as compared to tumors formed by vector control cells. Depletion of TFF3 in mammary carcinoma cells by siRNA concordantly decreased the angiogenic behavior of HUVEC. Forced Expression of TFF3 in mammary carcinoma cells stimulated IL-8 transcription and subsequently enhanced IL-8 Expression in both mammary carcinoma cells and HUVEC. Depletion of IL-8 in mammary carcinoma cells with Forced Expression of TFF3, or antibody inhibition of IL-8, partially abrogated mammary carcinoma cell TFF3-stimulated HUVEC angiogenic behavior in vitro, as did inhibition of the IL-8 receptor, CXCR2. Depletion of STAT3 by siRNA in MCF-7 cells with Forced Expression of TFF3 partially diminished the angiogenic capability of TFF3 on stimulation of cellular processes of HUVEC. Exogenous recombinant hTFF3 also directly promoted the angiogenic behavior of HUVEC. Hence, TFF3 is a potent angiogenic factor and functions as a promoter of de novo angiogenesis in mammary carcinoma, which may co-coordinate with the growth promoting and metastatic actions of TFF3 in mammary carcinoma to enhance tumor progression.
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trefoil factor 3 promotes metastatic seeding and predicts poor survival outcome of patients with mammary carcinoma
Breast Cancer Research, 2014Co-Authors: Vijay Pandey, Zhengsheng Wu, Min Zhang, Rui Li, Jian Zhang, Peter E. LobieAbstract:Introduction: Recurrence or early metastasis remains the predominant cause of mortality in patients with estrogen receptor positive (ER+) mammary carcinoma (MC). However, the molecular mechanisms underlying the initial progression of ER+ MC to metastasis remains poorly understood. Trefoil factor 3 (TFF3) is an estrogen-responsive oncogene in MC. Herein, we provide evidence for a functional role of TFF3 in metastatic progression of ER+ MC. Methods: The association of TFF3 Expression with clinicopathological parameters and survival outcome in a cohort of MC patients was assessed by immunohistochemistry. The Expression of TFF3 in MCF7 and T47D cells was modulated by Forced Expression or siRNA-mediated depletion of TFF3. mRNA and protein levels were determined using qPCR and western blot. The functional effect of modulation of TFF3 Expression in MC cells was determined in vitro and in vivo. Mechanistic analyses were performed using reporter constructs, modulation of signal transducer and activator of transcription 3 (STAT3) Expression, and pharmacological inhibitors against c-SRC and STAT3 activity. Results: TFF3 protein Expression was positively associated with larger tumour size, lymph node metastasis, higher stage, and poor survival outcome. Forced Expression of TFF3 in ER+ MC cells stimulated colony scattering, cell adhesion to a Collagen I-coated matrix, colony formation on a Collagen I- or Matrigel-coated matrix, endothelial cell adhesion, and transmigration through an endothelial cell barrier. In vivo, Forced Expression of TFF3 in MCF7 cells stimulated the formation of metastatic nodules in animal lungs. TFF3 regulation of the mRNA levels of epithelial, mesenchymal, and metastatic-related genes in ER+ MC cells were consistent with the altered cell behaviour. Forced Expression of TFF3 in ER+ MC cells stimulated phosphorylation of c-SRC that subsequently increased STAT3 activity, which lead to the downregulation of E-cadherin. siRNA-mediated depletion of TFF3 reduced the invasiveness of ER+ MC cells. Conclusions: TFF3 Expression predicts metastasis and poor survival outcome of patients with MC and functionally stimulates cellular invasion and metastasis of ER+ MC cells. Adjuvant functional inhibition of TFF3 may therefore be considered to ameliorate outcome of ER+ MC patients.
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artemin synergizes with twist1 to promote metastasis and poor survival outcome in patients with er negative mammary carcinoma
Breast Cancer Research, 2011Co-Authors: Arindam Banerjee, Peter E. Lobie, Jian Kang, Dong-xu Liu, Vijay Pandey, Tao Zhu, Pengxu QianAbstract:ARTEMIN (ARTN) is an estrogen regulated growth factor, the Expression of which promotes resistance to antiestrogen therapies and predicts poorer survival outcome of patients with estrogen receptor (ER) positive mammary carcinoma (ER+MC) treated with tamoxifen. ARTN is also expressed in ER negative mammary carcinoma (ER-MC). Herein, we determined the role of ARTN in ER-MC and defined the mechanism of action producing poor patient prognosis. We modulated the Expression of ARTN in two ER- (mesenchymal/claudin-low) mammary carcinoma cell lines (BT549 and MDA-MB-231) by Forced Expression or small interfering RNA (siRNA) mediated depletion. The effects of modulation of ARTN Expression were examined by various in vitro measures of oncogenicity, including the Expression of TWIST1 messenger RNA (mRNA) and protein. In vitro results were correlated to xenograft studies in immunodeficient mice. Co-Expression of ARTN and TWIST1 and their association to poor survival outcome were examined in a cohort of patients with ER-MC. Pathway analysis was performed by pharmacological inhibition of phosphorylation of AKT (pAKT-Ser 473) or modulation of TWIST1 Expression. ARTN Expression resulted in ER-MC cells with enhanced mesenchymal characteristics, including increased invasion and a gene Expression profile consistent with enhanced mesenchymal phenotype. ARTN stimulated ER-MC cell anchorage independent and 3D matrigel growth, endothelial cell adhesion and transmigration of ER-MC cells through an endothelial cell barrier. Forced Expression of ARTN produced a larger, locally invasive tumour mass with tumour emboli that produced distant metastasis. ARTN regulated TWIST1 Expression in ER-MC cells and ARTN Expression was significantly correlated to TWIST1 Expression in a panel of mammary carcinoma cell lines and in a cohort of patients with ER-MC. Low Expression of both ARTN and TWIST1 predicted 100% relapse free and overall survival in patients with ER-MC, whereas high Expression of both ARTN and TWIST1 was associated with a poor survival outcome. ARTN stimulated an increase in TWIST1 Expression via increased AKT activity. siRNA mediated depletion of TWIST1 abrogated ARTN stimulated cellular behaviour associated with metastasis, and Forced Expression of TWIST1 abrogated the functional effects of ARTN depletion. ARTN and TWIST1 synergize to produce a worse outcome in ER-MC and combined inhibition of ARTN and phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) may therefore provide a novel therapeutic strategy in this subtype of mammary carcinoma.