bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1075: Molecular determinants of colon and renal cancers' immune contextures
Authors: Becht Etienne , Giraldo Nicolas A. , Remark Romain , Reynies Aurelien de , Lacroix Laetitia , Damotte Diane , Sautès-Fridman Catherine , Fridman Wolf-Herman
Published: May 2015

In the vast majority of cancer types, as demonstrated and exemplified in colorectal cancer (CRC), a high density of CD8+ T cells in the tumor microenvironment correlates with a good prognosis for the patient. However, an opposite correlation has been reported in primary clear cell renal cell carcinoma (RCC) tumors. Our team studied the immune infiltrates of pulmonary metastases from CRC and RCC. We reported that despite the advanced stage of the disease in these patients, the immune infiltrate of pulmonary metastases remained a prognostic factor. As in the primary tumors, a high density of CD8+ T cells correlated with good prognosis for CRC metastases while it correlated with a bad prognosis for RCC metastases. These results suggest that the identity of the tumor cell is critical in shaping the immune contexture of a given tumor. We therefore investigated the molecular characteristics of the tumor cells which could be responsible for this opposite clinical impact in RCC versus CRC. First, we assessed the prognostic value associated with the expression of genes specific for different immune populations in several cohorts of primary CRC (n=177) and RCC tumors (n=72 and n=479), and confirmed the opposite impact of infiltrating lymphocyte densities on patient survival. In addition, RCC tumors showed a higher expression of genes related to inflammation, immunosuppression and angiogenesis. By comparing transcriptomic data from RCC and CRC cell lines, we observed that RCC tumor cells also expressed a significantly higher amount of these transcripts when compared to CRC tumor cells. Altogether, these results support the hypothesis that RCC malignant cells produce factors which are able to modify and determine the functional orientation of their immune microenvironment. Identifying which factors and pathways are critical for the RCC-related detrimental impact of CD8+ T cells on prognosis will be a major breakthrough, enabling the identification of new immunomodulatory drug targets and guide researchers and clinicians in immunotherapy protocols.

Citation Format: Etienne Becht, Nicolas A. Giraldo, Romain Remark, Aurelien de Reynies, Laetitia Lacroix, Diane Damotte, Catherine Sautès-Fridman, Wolf-Herman Fridman. Molecular determinants of colon and renal cancers' immune contextures. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1075. doi:10.1158/1538-7445.AM2014-1075
Abstract 1078: Cancer-associated fibroblasts promote endometrial cancer cell proliferation in vitro and in vivo
Authors: Chung Ivy , Subramaniam Kavita S. , Tham Seng Tian , Mohamed Zahurin , Adenan Noor Azmi Mat , Woo Yin Ling
Published: May 2015

Cancer-associated fibroblasts (CAFs) recently demonstrated tumor-promoting roles in various cancer types, yet their implication in endometrial cancer (EC) has not been fully explored. We isolated fibroblasts (CAFs) and its epithelial counterpart from human EC tissues using CD90- and CD326-antibodies conjugated magnetic beads, respectively. We collected CAFs secretion by concentrating spent supernatants from cells growing in media containing 2% fetal bovine serum. Both human EC cell lines and primary EC epithelial cells showed increased cell proliferation in a dose-dependent manner, following treatment with CAFs secretion. This was in contrast to the growth inhibitory effects shown with secretions from normal endometrial fibroblasts. EC cells also demonstrated increased cell motility and invasiveness in response to CAFs secretion. Using cytokine array and ELISA, we further identified several cytokines which were overexpressed in CAFs compared to normal endometrial fibroblasts: macrophage chemoattractant protein-1 (MCP-1), interleukin-6 (IL-6), growth regulated oncogene (GRO), regulated on activation, normal T cell expressed and secreted (RANTES) and vascular endothelial growth factor (VEGF). These cytokines may activate MAPK/Erk and/or PI3K/Akt pathways to promote EC cell proliferation, as shown by immunoblotting and ELISA. Indeed, CAFs-mediated EC proliferation was suppressed independently by specific inhibitors for PI3K/Akt (LY294002) and MAPK/Erk (U0126). To determine if CAFs promote EC tumor proliferation in vivo, we inoculated fluorescently labeled-epithelial cell and CAFs subcutaneously at the flank of nude mice in the ratio of 1:2 and 1:4. Tumor sizes were measured twice a week using calipers and monitored weekly using in vivo imaging. Inoculation of CAFs and normal fibroblasts alone did not induce any tumor growth; however, co-injection of EC with CAFs (1:4) showed approximately 3 times higher growth rate when compared to EC alone. More interestingly, there was no sign of tumor growth in mice inoculated with combination of EC and normal fibroblasts (1:4) at the end of experiment. Taken together, our data suggests that CAFs exert tumor-promoting effects both in vitro and in vivo partly via specific cytokines-mediated activation of MAPK/Erk and PI3K/Akt pathways. Delineating the roles of CAFs in EC tumor microenvironment may provide potential therapeutic targets for the treatment of EC.

Citation Format: Ivy Chung, Kavita S. Subramaniam, Seng Tian Tham, Zahurin Mohamed, Noor Azmi Mat Adenan, Yin Ling Woo. Cancer-associated fibroblasts promote endometrial cancer cell proliferation in vitro and in vivo. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1078. doi:10.1158/1538-7445.AM2014-1078
Abstract 1079: COX-2 blockade immunologically suppresses brain metastasis of lung cancer
Authors: Fujita Mitsugu , Nakata Susumu , Okuda Takeshi , Kato Amami , Yoshie Osamu
Published: May 2015

Epidemiological studies have highlighted associations between the regular use of nonsteroidal anti-inflammatory drugs (NSAIDs) and reduced cancer risks in human. In addition, NSAIDs reduce VEGF-D production levels, which in turn inhibit lymphatic vessel dilation and eventually cancer metastasis. Most NSAIDs function as cyclooxygenase-2 (COX-2) inhibitors that prevent the production of prostaglandin E2 (PGE2). The PGE2 induces myeloid-derived suppressor cells (MDSCs) in the bone marrow of cancer-bearing hosts, which is known to be responsible for T-cell suppression in chronic inflammation. Based on these findings, we hypothesized that COX-2 blockade would suppress brain metastasis of cancers by inhibiting MDSC development and their chemokine-mediated accumulation in the tumor microenvironment (TME). In both mouse and human brain metastasis cases, the expression levels of COX-2 and CCL2, a chemokine primarily attracting MDSCs, were increased in the brain tumor tissues compared with normal brain or peripheral bloods. Concomitantly, the infiltration of MDSCs (CD11+Gr-1+ for mice and CD14-CD15+HLA-DRlow for human, respectively) was observed at high levels in the tumor tissues. In the mouse brain tumor models using LL/2 murine lung cancer cell line, treatment with NSAIDs inhibited the PGE2 production levels and the formation of brain metastases. Furthermore, the NSAID treatment reduced the CCL2 in the TME, which was associated with a reduction in Ly6ChighLy6Glow monocytic and Ly6ClowLy6Ghigh granulocytic MDSCs in both the bone marrow and the TME. Mice genetically deficient for CCR2, a receptor for CCL2, exhibited low penetration of brain metastases along with reduced infiltration of MDSCs. Collectively, our findings show that the COX-2 pathway promotes brain metastases of lung cancers by supporting systemic development of MDSC and their accumulation in the TME mediated by the CCL2-CCR2 chemokine axis.

Citation Format: Mitsugu Fujita, Susumu Nakata, Takeshi Okuda, Amami Kato, Osamu Yoshie. COX-2 blockade immunologically suppresses brain metastasis of lung cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1079. doi:10.1158/1538-7445.AM2014-1079
Abstract 11: Absence of TGFBI favors tumor angiogenesis
Authors: Seo Jun-Young , Jung Hae Uk , Son Hye-Nam , Kim Soyoun , Jun Eunsung , Nam Ju-Ock , Kim Jung-Eun , Kim In-San
Published: May 2015

TGFBI, a circulating extracellular matrix protein, modulates several integrin-mediated cellular functions. The protein contains an N-terminal EMI domain, four consecutive FAS1 domains, and the RGD motif in the C-terminal domain. The fourth FAS1 domain of TGFBI exerts anti-angiogenic activity through interactions with αvβ3 integrin. Here, we used TGFBI-deficient mice and TGFBI transgenic mice to investigate the role of TGFBI in tumor growth in vivo. The absence of TGFBI led to enhanced tumor angiogenesis and a 3.6-fold increase in tumor growth. In contrast, a 1.7-fold increase in circulating levels of TGFBI in TGFBI transgenic mice induced a 2.4-fold decrease in tumor growth. We found that the binding affinity (Kd) of TGFBI for αvβ3 integrin was approximately 3.8 × 10 8 M, a value ∼2300-fold higher than that of a single FAS1 domain, and demonstrated that this greater affinity was due to the cooperative action of four FAS1 domains and the RGD motif. Moreover, TGFBI exhibited potent anti-angiogenic and anti-tumorigenic activities and specifically targeted the tumor vasculature and accumulated at the tumor site. Collectively, our results support the theory that TGFBI acts as a potent endogenous anti-tumor and anti-angiogenic molecule by targeting αvβ3 integrin, and highlight the importance of physiological circulating TGFBI levels in inhibiting tumor growth.

Citation Format: Jun-Young Seo, Hae Uk Jung, Hye-Nam Son, Soyoun Kim, Eunsung Jun, Ju-Ock Nam, Jung-Eun Kim, In-San Kim. Absence of TGFBI favors tumor angiogenesis. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 11. doi:10.1158/1538-7445.AM2014-11
Abstract 1103: MDSC targeting therapy for uterine cervical cancer displaying TRL
Authors: Matsumoto Yuri , Kawano Mahiru , Sasano Tomoyuki , Mabuchi Seiji
Published: May 2015

Objectives: The aim of this study was to investigate the clinical implications and underlying mechanism of tumor-related-leukocytosis (TRL) in cervical cancer. Methods: The clinical data of cervical cancer patients who were treated with definitive radiotherapy were analyzed to investigate the association between TRL and treatment outcome in retrospective and prospective settings. Clinical samples, cervical cancer cell lines, and a mouse model of cervical cancer were employed to examine the mechanisms responsible for TRL in cervical cancer, focusing on the myeloid-derived suppressor cell (MDSC) and tumor-derived granulocyte colony-stimulating factor(G-CSF). Results: TRL was significantly associated with shorter survival. The TRL-positive patients displayed upregulated tumor G-CSF expression and elevated serum G-CSF levels and increased MDSC frequencies in the blood compared with the TRL-negative patients. In vitro and in vivo investigations revealed that tumor-derived G-CSF is involved in the underlying causative mechanism of TRL and that the increase in the number of MDSC induced by tumor-derived G-CSF is responsible for the rapidly progressive nature of TRL-positive cervical cancer. The administration of anti-Gr-1 antibody or the depletion of MDSC by splenectomy significantly inhibited tumor growth in TRL-positive cervical cancer. Conclusion: TRL is an indicator of a poor prognosis in cervical cancer patients treated with definitive radiotherapy. MDSC induced by tumor-derived G-CSF is responsible for the rapidly progressive nature of TRL-positive cervical cancer. MDSC-targeting treatments might have therapeutic potential in TRL-positive cervical cancer.

Note: This abstract was not presented at the meeting.

Citation Format: Yuri Matsumoto, Mahiru Kawano, Tomoyuki Sasano, Seiji Mabuchi. MDSC targeting therapy for uterine cervical cancer displaying TRL. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1103. doi:10.1158/1538-7445.AM2014-1103
Abstract 1110: Colorectal liver metastases induce a systemic phase shift of the biological clock
Authors: Huisman Sander A. , Tamanini F. , Ahmadi A. , IJzermans J.N.M. , Horst G.T.J. van der , Bruin R.W.F. de
Published: May 2015

Circadian clock genes drive cyclic expression of 10-15% of the transcriptome. Disruption of the circadian clock may deregulate the cell's normal functions and can lead to carcinogenesis, indicating that clock genes also act as tumor suppressor genes. In previous work, we have shown that circadian rhythmicity is disrupted in C26 colon carcinoma cell-derived hepatic metastases, as compared to the non-cancerous tissue of the liver. It has been suggested that neoplastic tissue can influence the circadian clock of adjacent tissues, causing a tissue specific phase shift. A better understanding of how tumors affect the circadian system may help elucidate the role of the clock in cancer patients suffering from fatigue and sleep disorders. Therefore, we have examined the circadian clock in liver and kidney tissue of mice carrying hepatic C26 colon carcinoma cell metastases.

Male BALB/c mice were divided into two groups. In group 1, hepatic metastases were induced by injecting C26 colorectal carcinoma cells into the spleen. Group 2 was sham operated. Three weeks after (sham) surgery, animals were sacrificed over a 24-hour period (6 time points, n=4 per time point) and liver and kidneys were collected to compare circadian rhythmicity in control and tumor bearing animals. RNA was isolated and qRT-PCR was performed to determine the expression levels of 5 clock genes (Rev-Erbα, Per1, Per2, Bmal1, Cry1) and 3 clock controlled genes (Dbp, p21 and Wee1).

Liver and kidney tissue of healthy control mice showed normal 24-hour oscillations of all clock genes, consistent with normal circadian rhythmicity. Liver and kidney tissue of tumor-bearing mice, as compared to control mice, showed a marked phase difference in clock rhythms. In the liver we observed a 4-hour phase advance for Bmal1, Rev-Erbα and Dbp. In contrast in the kidney, there was a 4-hour phase delay for Bmal1, Rev-Erbα, Cry1 and DBP, and a 8 hour delay for Per2.

These data show that colorectal liver metastases induce a phase advance in the liver, and a phase delay in the kidney . This suggests a systemic effect of the tumor on peripheral clocks. This difference in clock phase between tumor-bearing mice and healthy controls may provide new clues to further understand the role of the clock for fatigue and sleep problems in cancer patients, but may also give opportunities to administer chemotherapeutic treatment in a chronotherapeutical setting .

Citation Format: Sander A. Huisman, F. Tamanini, A. Ahmadi, J.N.M. IJzermans, G.T.J. van der Horst, R.W.F. de Bruin. Colorectal liver metastases induce a systemic phase shift of the biological clock. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1110. doi:10.1158/1538-7445.AM2014-1110
Abstract 1112: Role of osteoprotegerin in inflammatory and invasive breast cancer
Authors: Goswami Sudeshna , SHARMA-WALIA NEELAM
Published: May 2015

Inflammatory breast cancer (IBC) is a highly aggressive and angioinvasive form of breast cancer associated with a high incidence of early nodal and systemic metastasis. A proactive, complex and dynamic tumor microenvironment of IBC adds to the grim scenario of the disease by accumulating inflammatory and angiogenic growth factors and creating a niche for the growth and metastasis of tumor cells. The secretions from the primary inflammatory ductal carcinoma of the breast (SUM149PT), and highly invasive ductal carcinoma cells (SUM1315MO2) secrete unusually high levels of osteoprotegerin (OPG) as compared to those obtained from the primary human mammary epithelial cells (HMEC). OPG, which plays a pivotal role in bone remodeling by regulating osteoclast formation, is a multifaceted molecule, and has been found to be involved in multiple myeloma and prostate cancers. This prompted us to examine the role of OPG in the aggressive breast cancers, which has not been deciphered till date. For a better understanding of the OPG functions, apart from using adherent cells, we used in vitro ‘sphere culture’ as our working model. We studied the functional significance of OPG in aggressive breast cancer biology by utilizing well standardized techniques such as OPG staining (immunohistochemistry, immunofluorescence), OPG secretion (ELISA), angiogenesis (in vitro tube formation), proliferation (MTT), and cell cycle/aneuploidy analysis (PI staining). We demonstrated that (1) OPG secretion as well as protein level is upregulated in IBC (SUM149PT) and highly Invasive ductal carcinoma cells (SUM1315MO2), and their spheres compared to those of normal human mammary epithelial (HMEC); (2) OPG stained specifically in patient breast cancer tissue samples when compared to the uninvolved tissue from the same patient; (3) OPG rich secretions from IBC and highly invasive breast cancer were highly angiogenic; (4) OPG enriched culture medium induced proliferation of the control HMEC spheres; (5) the presence of OPG triggered the onset of aneuploidy in the control HMEC adherent cells as well as spheres. The functional studies showing the biological significance of OPG (i,e its effect on proliferation, angiogenesis, and aneuploidy) were confirmed using recombinant human OPG and OPG rich/depleted medium obtained from breast cancer cells. Overall our studies highlight the link between OPG and aneuploidy in aggressive breast cancer cells, and opens up avenues for new therapeutic targets for treating IBC.

Citation Format: Sudeshna Goswami, NEELAM SHARMA-WALIA. Role of osteoprotegerin in inflammatory and invasive breast cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1112. doi:10.1158/1538-7445.AM2014-1112
Abstract 1117: Platelet interaction induces Tissue Factor and a pro-metastatic phenotype in ovarian cancer cells
Authors: Orellana Renan F. , Oliva Barbara , Kato Sumie , Bravo Loreto , Gonzalez Pamela , Panes Olga , Mezzano Diego , Cuello Mauricio , Owen Gareth
Published: May 2015

Introduction: An increase in circulating platelets, or thrombocytosis, is recognized as an independent risk factor correlated with bad prognosis and metastasis, however the exact role that platelets play in tumor progression has not been fully elucidated. The expression of Tissue Factor (TF) protein by cancer cells has been shown to correlate with Hypercoagulable state and metastasis. It is speculated that cancer cells need to acquire a “Metastasis Initiating Cell (MIC)” phenotype. To achieve this, cells may suffer an Epithelial to Mesenchymal Transition (EMT) coupled with stem-like protein expression in which E-cadherin is lowered, while CD44 and N-cadherin are increased. Ovarian cancer patients develop ascitis, a build up of peritoneal fluid that contains cancerous and serum derived cells. After the surgical removal of the primary and visible tumors, these ascitic borne cells form peritoneal tumors and thus make a good model to study platelet/cancer cells interaction in metastasis. The aim of this work was determine the effect of platelet-cancer cell interaction on TF and EMT marker levels and cancer cell migration in ascites-derived cancer cells. Material and Methods: Ovarian ascitic-isolated cancer cells and cell lines were co-cultivated with human washed platelets (from healthy donors). TF and EMT marker levels were determined by Western blotting. Cancer cell migration was determined by Boyden chambers and the scratch assay. Results: The co-culture of ovarian cancer cells with platelets causes: 1) a phenotypic change in cancer cells, 2) increases TF protein levels and activity, 3) induces EMT markers, 3) promotes chemoattraction and cancer cell migration. Conclusion: We present evidence that platelet-cancer cell interaction promotes a Metastasis Initiating Cell phenotype and enhanced migration. We speculate that platelet-cancer cell interaction may promote the formation of metastatic foci worsening the patient prognosis.

Fundings: FONDECYT 1120292, 3120003, 1100870, CORFO 13IDL2-18608, BMRC 13CTI-21526-P6

Citation Format: Renan F. Orellana, Barbara Oliva, Sumie Kato, Loreto Bravo, Pamela Gonzalez, Olga Panes, Diego Mezzano, Mauricio Cuello, Gareth Owen. Platelet interaction induces Tissue Factor and a pro-metastatic phenotype in ovarian cancer cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1117. doi:10.1158/1538-7445.AM2014-1117
Abstract 1118: Stress-induced differential Survivin release in prostate cancer health disparities
Authors: Khan Salma , Turay David , Jutzy Jessica Ms , Aspe Jonathan R. , Sanchez Tino W. , Mirshahidi Saeid , Casiano Carlos A. , Wall Nathan R.
Published: May 2015

African-American men have the highest rate of prostate cancer (PCa) incidence in the U.S. In addition, their prostate cancer mortality rate is more than twice as high as the rate for white Americans. The causes of higher rates of prostate cancer among African-American males are largely unknown. Many studies are underway to unravel the impact of a wide variety of potential risk factors, including dietary and other lifestyle differences, occupational exposures, hormonal and genetic differences. Several lines of evidence suggest that one of the main events associated with the conversion to an aggressive phenotype is increased resistance to apoptosis (3) mainly due to upregulation of antiapoptotic genes, including Bcl-2, Bcl-XL, Mcl-1, and Survivin. Recently, we reported that Survivin is released in PCa sera and found to be in the exosomes. Therefore, we speculate that Survivin could be a potential target protein released differentially in prostate cancers in African American (AA) and Caucasian (CC) populations in response to stress. In this study, we have collected serum samples from prostate cancer patients of AA and CC, measured Survivin levels by ELISA. Exosome islation was perfomed by Exoquick approach and quantified by acetylcholinesterase activity assays. We have applied an in-vitro approach to compare the differences between cancer cell lines derived from AA (MDA-MB-PCa) and CC (PC3) patients in terms of stress and stress activated Survivin release by exosomes. Under oxidative stress and drug treatments, we have identified Survivin highly expressed in exosome derived from MDA-MB-PCa compared to PC3 cell lines. In addition, we found a relative overexpression of Survivin protein in AA-PCa compared to CC-PCa both in sera and exosomes. Based on our data, Survivin may be a key stress-induced protein in AA-PCa patients. It is possible that attempts to block Survivin release will not only interrupt the cancer proliferation and interfere with anti-apoptotic pathways, thus preventing cancer progression.

Citation Format: Salma Khan, David Turay, Jessica Ms Jutzy, Jonathan R. Aspe, Tino W. Sanchez, Saeid Mirshahidi, Carlos A. Casiano, Nathan R. Wall. Stress-induced differential Survivin release in prostate cancer health disparities. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1118. doi:10.1158/1538-7445.AM2014-1118
Abstract 1121: Extracellular matrix-integrin B1 signaling is a major mediator of epithelial-to-mesenchymal transition and contributes to prostate cancer invasion and metastasis
Authors: Bugiel Steven R. , McKittrick Elisabeth , Zhao Huijun , Howe Grant A. , Addison Christina L.
Published: May 2015

Within North America, prostate cancer is the most commonly diagnosed cancer in men with devastating implications deriving from metastasis, particularly those established in the bone. We have elucidated mechanisms by which prostate cancer undergoes metastasis with particular focus on the relationship between extracellular-matrix binding proteins, Integrin B1 (ITGB1) and epithelial-to-mesenchymal transition (EMT). Previous results in the laboratory suggested that prostate tumor cells depleted of ITGB1 were unable to form colonies in soft agarose and were impaired in their invasive abilities in vitro. In order to determine the possible mechanisms by which ITGB1 controlled these phenotypes, we performed numerous genetic techniques including siRNA targeted depletion of ITGB1 and subsequent profiling of pathway specific message RNA by RTqPCR to monitor the effects ITGB1 depletion on factors known to control EMT which is also associated with cell invasion. We found that prostate tumor cells depleted of ITGB1 had altered expression of numerous genes associated with EMT or cell invasion, many of which have been validated by RT-qPCR and western blot. In particular, a protease inhibitor SERPINE1 also known as Plasminogen Activator Inhibitor 1 (PAI-1) which is associated with EMT has been established as a target of ITGB1 regulation, in part via the ability of ITGB1 to regulate upstream factors associated with SERPINE1 expression such as TGFB2. Of particular interest to prostate cancer bone metastasis, we further found that collagen I interacting with ITGB1 significantly upregulated this pathway, and given the known role of EMT in mediating resistance to anti-cancer agents, this finding could help elucidate novel targets for the development of new drugs to treat prostate cancer bone metastasis.

Citation Format: Steven R. Bugiel, Elisabeth McKittrick, Huijun Zhao, Grant A. Howe, Christina L. Addison. Extracellular matrix-integrin B1 signaling is a major mediator of epithelial-to-mesenchymal transition and contributes to prostate cancer invasion and metastasis. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1121. doi:10.1158/1538-7445.AM2014-1121
Abstract 1122: Tetrandrine induces expression of growth/differentiation factor-15 and promoted terminal differentiation of PCa cells
Authors: Koul Sweaty , Koul Hari K.
Published: May 2015

Tetrandrine (Tet) is a naturally derived bis-benzylisoquinoline alkaloid, from S tenrandra, a herb that has been used in Chinese medicine for its anti-inflammatory, immunologic and antiallergenic effects. Results from our laboratory show that Tetrandarine blocks prostate cancer growth, promotes cell cycle arrest and induces apoptosis in a time and concentration dependent manner. In the present study we performed un-biased gene expression analysis of LNCaP cells following treatment with Tet. Analysis of gene expression revealed that GDF-15 was one of the most highly expressed cytokines following treatment of LNCaP cells with Tet. The gene expression results were further confirmed in relative quantitative PCR and Real time PCR assays, as was protein expression of GDF-15 confirmed by Western blot analysis. Our results, in light of the role of GDF-15 as a macrophage inhibitory factor as well as prostate epithelial differentiating factor, suggest that Tet may serve as a useful agent to promote prostate epithelial differentiation and inhibit tumor macrophage infiltration. Additional studies revealed that Treatment of PCa cells with Tet inhibited cell migration and invasion and suppressed EMT. Taken together we conclude that Tet by regulating GDF-15 expression may serve as an inhibitor of Prostate cancer cell progression and metastasis.

(Financial Support: Supported in part by NIH/NCI R01CA161880 (PI:H.Koul); VA Merit Award 01BX001258 (PI:H.Koul); Carroll W. Feist endowed Cahir and LSUHSC Chair Commitment Funds)

Citation Format: Sweaty Koul, Hari K. Koul. Tetrandrine induces expression of growth/differentiation factor-15 and promoted terminal differentiation of PCa cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1122. doi:10.1158/1538-7445.AM2014-1122
Abstract 1123: Fucosylated TGF-ß receptors transduce a signal for epithelial-mesenchymal transition in colorectal cancer cells
Authors: Takimoto Rishu , Hirakawa Masahiro , Tamura Fumito , Yoshida Makoto , Ono Michihiro , Sato Yasushi , Osuga Takahiro , Kato Junji
Published: May 2015

Background: Transforming growth factor-ß (TGF-ß) is a major inducer of epithelial-mesenchymal transition (EMT) in different cell types. TGF-ß-mediated EMT is thought to contribute to tumor cell spread and metastasis. Sialyl Lewis antigens synthesized by fucosyltransferase (FUT) 3 and FUT6 are highly expressed in patients with metastatic colorectal cancer (CRC) and are utilized as tumor markers for cancer detection and evaluation of treatment efficacy. However, the role of FUT3 and FUT6 in augmenting the malignant potential of CRC induced by TGF-ß is unclear.

Methods: CRC cell lines were transfected with siRNAs for FUT3/6 and were examined by cell proliferation, invasion, and migration assays. The expression and phosphorylation status of TGF-ß downstream molecules were analyzed by western blot. Fucosylation of TGF-ß receptor was examined by lectin blot analysis.

Results: Inhibition of FUT3/6 expression by siRNAs suppressed the fucosylation of type I TGF-ß receptor and phosphorylation of the downstream molecules, thereby inhibiting the invasion and migration of CRC cells by EMT.

Citation Format: Rishu Takimoto, Masahiro Hirakawa, Fumito Tamura, Makoto Yoshida, Michihiro Ono, Yasushi Sato, Takahiro Osuga, Junji Kato. Fucosylated TGF-ß receptors transduce a signal for epithelial-mesenchymal transition in colorectal cancer cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1123. doi:10.1158/1538-7445.AM2014-1123
Abstract 1126: Tristetraprolin inhibits Twist1-induced cancer cell migration
Authors: Cho Wha J. , Yoon Nal A. , Vo Mai-Tram , Min Young Joo , Park Jeong Woo
Published: May 2015

Epithelial-Mesenchymal Transition (EMT) enhances tumor cell migration. However, detail mechanisms for the regulation of EMT are not well known. Tristetraprolin (TTP) is an AU-rich element (ARE) binding protein that can promote the decay of the transcripts. This study demonstrates that TTP plays a critical role in EMT process. Overexpression of TTP in ovarian cancer cells decreased the expression of mesenchymal markers (N-cadherin and Vimentin), increased the expression of epithelial marker (E-cadherin), and inhibited cell migration and invasion activities. On the contrary, inhibition of TTP by siRNA decreased the expression of E-cadherin, increased the expression of N-cadherin and Vimentin, and promoted cell migration and invasion. The cDNA microarray analysis revealed that Twist1 was significantly down-regulated in TTP-overexpressed cells. Twist1 mRNA contains ARE within its 3′-UTR and TTP enhances the decay of Twist1 mRNA through binding to its 3′-UTR. In addition, TTP directly bound to the ARE of Twist1 mRNA by EMSA assay. Ectopic expression of Twist1 gene without 3′UTR blocked the inhibitory of TTP on the EMT. We also observed enhanced levels of Twist1 and reduced TTP levels in human ovarian adenocarcinoma. Collectively, our data indicate that TTP conducts anti-metastatic activity by down-regulating Twist1, presenting a novel mechanism of TTP-mediated regulation of tumor metastasis.

Citation Format: Wha Ja Cho, Nal Ae Yoon, Mai-Tram Vo, Young Joo Min, Jeong Woo Park. Tristetraprolin inhibits Twist1-induced cancer cell migration. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1126. doi:10.1158/1538-7445.AM2014-1126
Abstract 1127: Snail inhibits lung colonization and metastasis in luminal type A breast cancer cells
Authors: Lin Shih-Pei , Hung Shih-Chieh
Published: May 2015

Breast cancer is the most common malignant disease in Western women. In these patients, it is not the primary tumor, but its metastases at distant sites that are the main cause of death. Breast cancer include five biologically distinct subgroups including Luminal A (ER+ ck8h/ck18h), Luminal B (ER+ ck8l/ck18l), Basal (ER- ck5+/ck17+) and ERBB2+ (ER- ERBB2+ GFRBP7+) and normal breast-like types. The time to distant metastasis and overall survive rate were distinct among these subgroups. Luminal type has the best prognosis, while basal type or ERBB2+ has the worst prognosis. What mechanism involves in this difference remains to be elucidated. In the current study, we demonstrated (1) Luminal type A tumor such as MCF7 when overexpressed with active mutant form of Snail gene increased in the expression of markers for epithelial mesenchymal transition such as Vimentin, N-cadherin and Fibronectin but decreased in the expression of E-cadherin compared to cells overexpressed with control vectors of wild type Snail gene. Moreover, these cells increased in the capacity of migration and invasion. (2) While, MCF7 overexpressed with active mutant form of Snail decreased in the ability to survive and form sphere in serum-free medium, the in vitro colonization, and were unable to metastasize to lung.Therefore, the elucidation of the differences in molecules or signaling pathways involve in controlling migration, invasion and distant metastasis may have a therapeutically beneficial effect on breast cancer treatment.

Citation Format: Shih-Pei Lin, Shih-Chieh Hung. Snail inhibits lung colonization and metastasis in luminal type A breast cancer cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1127. doi:10.1158/1538-7445.AM2014-1127
Abstract 1133: Partial EMT, hybrid epithelial-mesenchymal phenotype and epithelial plasticity: role of (miR-200/ZEB) loop
Authors: Jolly Mohit Kumar , Lu Mingyang , Levine Herbert , Onuchic Jose N. , Ben-Jacob Eshel
Published: May 2015

Epithelial to Mesenchymal Transition (EMT) and Mesenchymal to Epithelial Transition (MET) are critical for embryonic development and tissue repair. Aberrantly regulated transitions are also implicated in cancer metastasis. The core regulatory genetic network for these transitions appears to allow for the existence of a hybrid epithelial/mesenchymal phenotype (E/M). Such hybrid cells are seen during wound healing and branching morphogenesis, and have mixed epithelial and mesenchymal characteristics, which enable them to migrate collectively. The core regulatory network for the cell fate determination between these three phenotypes is composed of two highly interconnected mutual-inhibition feedback circuits - miR-34/SNAIL and miR-200/ZEB. Here we applied a novel theoretical framework to model the microRNA-based regulation to study this core regulatory unit. More specifically, we looked into the functions of the two loops in isolation, and subsequently of the combined unit when the two loops are integrated into the full regulatory circuit. We found that miR-200/ZEB forms a tristable circuit that acts as a ternary decision making switch, driven by miR-34/SNAIL that is a monostable loop which acts as noise-buffering integrator of internal and external signals that induce or inhibit EMT/MET. Our results are corroborated by recent experiments indicating the (miR-200/ZEB) to be the commitment point and decision making loop (eg. Cells don't revert to epithelial phenotype on removal of EMT-inducing signal, if ZEB expression has been turned on.) .We associate the three stable states of the miR-200/ZEB circuit to different phenotypes - (1,0) - (high miR-200)/(low ZEB) - epithelial, (0,1) - (low miR-200)/(high ZEB) - mesenchymal, and (1/2,1/2) - (medium miR-200)/(medium ZEB) - hybrid. Thus, we propose that the hybrid epithelial - mesenchymal phenotype has concurrent expression of both miR-200 and ZEB. Our hypothesis is consistent with recent experimental studies (e.g. ZEB expression measurements in collectively migrating cells), and also explains the lack of hybrid phenotype observed during MET in colonization and induced pluripotent stem cells (iPSCs). We also present testable predictions of dynamic gene expression during complete and partial transitions (EMT/MET).

Citation Format: Mohit Kumar Jolly, Mingyang Lu, Herbert Levine, Jose N. Onuchic, Eshel Ben-Jacob. Partial EMT, hybrid epithelial-mesenchymal phenotype and epithelial plasticity: role of (miR-200/ZEB) loop. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1133. doi:10.1158/1538-7445.AM2014-1133
Abstract 1137: Bub1 is a key regulator of TGF-β signaling
Authors: Nyati Shyam , Schinske-Sebolt Katrina , Pitchiaya Sethuramasundaram , Chekhovskiy Katerina , Chator Areeb , Chaudhry Nauman , Dosch Joseph , Dort Marcian E. Van , Varambally Sooryanarayana , Kumar-Sinha Chandan , Nyati Mukesh K. , Ray Dipankar , Walter Nils G. , Yu Hongtao , Ross Brian D. , Rehemtulla Alnawaz
Published: May 2015

The transforming growth factor-beta (TGF-β) family of cytokines regulates many processes such as immune suppression, angiogenesis, wound healing and epithelial to mesenchymal transition (EMT. Early in tumorigenesis, when epithelial cells retain exquisite growth sensitivity to this ligand, TGF-β signaling elicits a tumor suppressing activity. However, transformed cells become refractory to TGF-β-mediated growth inhibition and acquire a phenotype wherein the intracellular signaling circuitry of the cells is altered, leading to tumorigenic and metastatic effects in response to TGF-β exposure. Although TGF-β activating pathways have been studied, the molecular participants are poorly defined. Here we identify budding uninhibited by benzimidazoles-1 (Bub1) as an integral component of canonical and non-canonical TGF-β signaling pathways, where Bub1 is required for TGFBRI-TGFBRII complex formation and activation. Bub1-depleted cells exhibited reductions in TGF-β dependent Smad2/3 phosphorylation, recruitment of Smad2/3 to the TGFBRI-II complex, PI3K/Akt and p38MAPK activation, Smad binding element driven promoter activity (SBE4-Luc), and invasion and migration. Furthermore, a targeted small molecule inhibitor of Bub1 kinase activity (2OH-BNPP1), as well as an inactive kinase mutant of Bub1, abrogated ligand mediated TGF-β signaling and phenotypic response. These studies demonstrate a role for the Bub1 kinase in mediating TGF-β dependent signaling beyond its established function in cell-cycle regulation and chromosome cohesion and uncover the underlying basis for the pleiotropic cellular response commonly observed upon activation of the pathway.

Citation Format: Shyam Nyati, Katrina Schinske-Sebolt, Sethuramasundaram Pitchiaya, Katerina Chekhovskiy, Areeb Chator, Nauman Chaudhry, Joseph Dosch, Marcian E. Van Dort, Varambally, Kumar-Sinha, Nyati, Ray, Walter, Sooryanarayana Varambally, Chandan Kumar-Sinha, Mukesh K. Nyati, Dipankar Ray, Nils G. Walter, Hongtao Yu, Brian D. Ross, Alnawaz Rehemtulla. Bub1 is a key regulator of TGF-β signaling. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1137. doi:10.1158/1538-7445.AM2014-1137
Abstract 1145: Cytoplasmic p27 promotes epithelial-mesenchymal transition and tumor progression via Twist1 upregulation
Authors: Zhao Dekuang , Besser Alexandra H. , Zhou Wen , Wander Seth A. , Sun Jun , Durante Michael , Hong Feng , Wang Bin , Ince Tan , Briegel Karoline , Slingerland Joyce M.
Published: May 2015

p27 is a cell cycle regulator that acts largely to restrain normal cell growth. It is rarely mutated or deleted in human cancers, but is often degraded or mislocalized to the cytoplasm in aggressive tumors. Phosphorylation at T157 or T198 by different PI3K effector kinases leads to p27 cytoplasmic accumulation and enhanced cell motility and invasion in part via effects on the actin cytoskeleton, which are independent of its cell cycle role. However, the functional contributions of C-terminally phosphorylated, cytoplasmic p27 to cancer progression remain poorly understood. Targeted inhibition of PI3K/mTOR impaired tumor cell motility and metastasis via modulation of p27 in a bone metastatic model (Wander, S. et al. BCRT 2013). We observed that p27 knockdown in highly metastatic cancer lines with high levels of cytoplasmic p27pT157pT198 reverted EMT and impaired tumor cell invasion in vitro and metastasis in vivo. This led us to investigate whether PI3K-activated, phosphorylated p27 may function as a novel EMT regulator. A cell cycle defective (CK-) and double phosphomimetic p27 mutant (T157D/T198D or DD) was introduced into immortal human mammary epithelial cells with low PI3K activity. This revealed a novel, oncogenic function of p27 in regulating tumor progression, in that p27CK-DD induced epithelial-mesenchymal transition (EMT) and soft agar colony formation. p27CK-DD also increased motility, invasion and formation of metastasis in bladder and breast cancer models. A RT-PCR screen for potential p27-induced EMT mediators revealed a 20-fold increase in expression of the Twist1 transcription factor in p27CK-DD-transduced cells. Knockdown of Twist reversed the p27CK-DD-mediated increase in EMT marker expression. p27 knockdown rapidly attenuated Twist1-promoter activity and reduced Twist1 mRNA levels and also increased E-cadherin expression in metastatic cells, suggesting that Twist1 may play a critical role in p27CK-DD-induced EMT. These data extend our understanding of p27 function in human cancer and suggest that PI3K deregulated p27 may promote EMT and tumor metastasis through transcriptional activation of TWIST1.

Citation Format: Dekuang Zhao, Alexandra H. Besser, Wen Zhou, Seth A. Wander, Jun Sun, Michael Durante, Feng Hong, Bin Wang, Tan Ince, Karoline Briegel, Joyce M. Slingerland. Cytoplasmic p27 promotes epithelial-mesenchymal transition and tumor progression via Twist1 upregulation. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1145. doi:10.1158/1538-7445.AM2014-1145
Abstract 1153: Stromal FoxM1 and p53 in lung cancer as mediators of tumor progression, metastatic spread and chemo-resistance
Authors: Kamer Iris , Daniel Inbal , Perry Gili , Onn Amir , Bar Jair
Published: May 2015

Introduction:

Insight into the processes of tumor initiation and progression as well as metastatic spread and response to therapies requires studies that take into consideration the stromal and epithelial components of tumors. Both FoxM1 and p53 have a major role in the progression, metastasis and chemo-resistance of non-small cell lung cancer (NSCLC). p53 is a well-recognized tumor-suppressor gene, commonly mutated in NSCLC and correlated with tumor aggressiveness. An anti-tumor role of p53 in the stroma has been demonstrated by us and others. p53 is reported to suppress FoxM1 transcription. FoxM1 is an oncogene, contributing to lung cancer progression and metastatic spread. Little is known about the role of FoxM1 in cancer's micro-environment. We hypothesized that tumor-induced-downregulation of p53 and upregulation of FoxM1 in the stromal compartments promotes lung cancer progression, metastatic spread and chemo-resistance.

Materials and Methods:

Cancer associated fibroblasts (CAFs) and normal fibroblasts (NFs) were produced from fresh lung tumor specimens. Cell lines and primary cells were grown in standard tissue culture conditions. Co-cultures were performed using transwell culture systems. DNA damage was induced by cisplatin treatment. Expression levels of mRNA and proteins were tested by real-time RT-PCR and western blots.

Results:

We have found FoxM1 levels to be higher in primary lung CAFs compared to NFs. In addition, CAFs that were treated with conditioned media of transformed epithelial cells showed down-regulation of stromal-p53 and up-regulation of FoxM1, in DNA damage conditions. mRNA levels of FoxM1 remain stable. While cancer-cells-CM up-regulate FoxM1 in CAFs, it was found to down-regulates it in NFs, suggesting both cancer and CAFs have evolved a unique mechanism of FoxM1 induction in stroma.

In order to examine a possible paracrine effect between p53 levels in the stroma, and FoxM1 levels in tumor cells, we co-cultivated fibroblasts knocked-down for p53 with lung cancer cells and found up-regulation of FoxM1 the tumor cells.

Summary:

We demonstrate up-regulation of stromal FoxM1 by cancer-secreted-factors as well as by stable CAFs modifications, concomitantly to stromal p53 down-regulation. High stromal FoxM1 may promote tumor cells aggressiveness, similarly to low stromal p53, possibly being downstream to p53. Elaboration of these pathways will deepen our understanding of lung cancer and suggest novel therapeutic targets.

Citation Format: Iris Kamer, Inbal Daniel, Gili Perry, Amir Onn, Jair Bar. Stromal FoxM1 and p53 in lung cancer as mediators of tumor progression, metastatic spread and chemo-resistance. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1153. doi:10.1158/1538-7445.AM2014-1153