bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1149: The role of the novel Notch1-Sox9 signaling axis in NSCLC progression and EMT
Authors: Capaccione Kathleen M. , Hong Xuehui , Morgan Katherine M. , Allen Thaddeus D. , Miles Gregory D. , Markert Elke K. , Bishop J. Michael , Pine Sharon R.
Published: May 2015

Sox9 plays critical roles in the specification and differentiation of numerous progenitor and differentiated cell types during embryonic and fetal development. Sox9 is overexpressed in 40 - 50% of lung adenocarcinomas and associated with poor prognosis in lung cancer patients. We set out to identify upstream pathways that regulate Sox9 expression in lung cancer, as well as the role of Sox9 in lung adenocarcinoma progression. Several developmental and stem cell pathways are known to induce Sox9 transcription during carcinogenesis, including the TGB-β, Wnt/β-catenin, Sonic Hedgehog, and NF-κB signaling. Sox9 has also been shown to be a transcriptional target of the Notch pathway during mouse development, although the binding sites for Notch within the mouse Sox9 promoter are not conserved in humans. We mined gene expression data from three publicly available datasets and found that Hes1, a known Notch target gene, is co-expressed with Sox9 in lung adenocarcinoma. Furthermore, Sox9 mRNA and protein levels were upregulated over 100-fold as early as 14 days after Notch1 induction in the Notch1-induced mouse model of lung cancer, suggesting that Sox9 overexpression is an early event during lung cancer development. Through a series of in vitro assays, we determined that Sox9 is downstream of Notch1 in lung adenocarcinoma cell lines. By ChIP we determined that Sox9 is a direct target of Notch1 and using luciferase reporter assays, we located the previously unidentified human RBP-Jκ binding site, the principle effector of canonical Notch1 signaling, immediately upstream of the Sox9 transcriptional start site. We also examined TGF-β, a known inducer of epithelial-to-mesenchymal transition (EMT) in lung cancer. We determined that induction of Sox9 expression by Notch1 is independent of TGF-β signaling and that TGF-β and Notch1 cooperate in their regulation of Sox9 expression. Loss of Notch1 expression led to an induced MET phenotype, characterized by decreased cell invasion/migration, MET-like morphological changes, and increased E-cadherin expression, which were rescued by Sox9 overexpression. Our data also demonstrate that Sox9 contributes to Notch1-induced EMT in lung adenocarcinoma. These results establish Sox9 as a key Notch1 target gene mediating Notch1-induced EMT independent of TGF-β, leading to poor survival in lung adenocarcinoma.

Citation Format: Kathleen M. Capaccione, Xuehui Hong, Katherine M. Morgan, Thaddeus D. Allen, Gregory D. Miles, Elke K. Markert, J. Michael Bishop, Sharon R. Pine. The role of the novel Notch1-Sox9 signaling axis in NSCLC progression and EMT. [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 1149. doi:10.1158/1538-7445.AM2014-1149
Abstract 1150: The role of the tumor suppressor gene THY1 in suppression of epithelial-mesenchymal transition (EMT) in nasopharyngeal carcinoma
Authors: Lung Hong Lok , Lung Maria Li
Published: May 2015

Background and aims:

THY1 has been successfully identified as a tumor suppressor gene (TSG) in nasopharyngeal carcinoma (NPC) in our laboratory. The frequency of down-regulated THY1 protein expression was found to be significantly associated with NPC lymph node metastases. However, direct functional evidence is still lacking for THY1 being anti-metastatic in NPC. Epithelial-Mesenchymal Transition (EMT) is well-known as a closely associated process for stem cells as well as tumor cells to invade to other part of the body. We aim to study the association of EMT with the THY1 in NPC.

Methods: Functional analysis of restoration of THY1 expression in the NPC cell lines was studied. The RhoA negative regulator, p190 Rho GTPase-activating protein (p190RhoGAP) and the E-cadherin/beta-catenin cell junction was examined in the THY1-expressing NPC cells.

Results: Both real-time and conventional invasion chamber assays clearly showed that the invasive ability of the THY1 transfectants was consistently lower than vector-alone control. The reverse transcription-polymerase chain reaction (RT-PCR) results show that the gene expression of cell invasion-associated gelatinase MMP-9 was significantly down-regulated in the THY1-transfectants compared with the vector-alone control. The p190RhoGAP was greatly activated by phosphorylation in the THY1 transfectants, when the THY1 gene was switched on in the absence of doxycycline (dox, an analogue of tetracycline). In the presence of dox when THY1 was switched off, the phosphorylation of p190RhoGAP was reduced. It appears that the phosphorylation status of p190RhoGAP is positively regulated by THY1 in NPC cells. The immunofluorescence (IF) confocal microscopy results show that beta-catenin and E-cadherin were much more frequently translocated to the cell-cell junction in the THY1 transfectants than the vector-alone.

Conclusions: These findings suggest that THY1 inhibited NPC cell invasion via the formation of adherens junction and up-regulation of p190RhoGAP. The enhanced formation of cell adherens junction by THY1 is likely to be one of the possible mechanisms to suppress EMT in NPC.

Citation Format: Hong Lok Lung, Maria Li Lung. The role of the tumor suppressor gene THY1 in suppression of epithelial-mesenchymal transition (EMT) in nasopharyngeal carcinoma. [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 1150. doi:10.1158/1538-7445.AM2014-1150
Abstract 1160: Novel CSF-1 receptor ligand IL-34 modulates macrophage-breast cancer cell crosstalk
Authors: Gunawardhana Sandun , Zins Karin , Lucas Trevor , Abraham Dietmar
Published: May 2015

Crosstalk between stromal cells and malignant cells within the tumor microenvironment is important for tumorigenesis. According to current clinical and experimental evidence, tumor associated macrophages (TAMs) play a major role in tumor progression to metastasis in different cancers. In breast cancer, increased number of TAMs is associated with poor prognosis. Previous work in our laboratory has shown that depletion of TAMs by blocking colony-stimulating factor-1 (CSF-1) suppresses tumor growth, matrix metalloprotease production and further recruitment of macrophages in a MCF-7 breast cancer xenograft model. In this study, we aim to investigate Interleukin-34 (IL-34), a novel ligand for the colony stimulating factor-1 receptor (CSF-1R). IL-34 functions as a specific and independent ligand and as an agonist to the CSF-1R. Furthermore it is known to induce proliferation through mitogen activated protein kinase (MAPK) activation in macrophages and cancer cells. To date, nothing is known about the role of IL-34 in breast cancer and its effects on downstream signalling pathway modulation in cancer cell-macrophage crosstalk. Our real-time PCR data shows that in co-cultures with murine macrophages, human MCF-7 and MDA-MB-231 breast cancer cells express increased levels of IL-34. In addition, in vitro migration assays demonstrated that recombinant human and murine IL-34 significantly increases the migration of murine macrophages, MCF-7 and MDA-MB-231 breast cancer cells. IL-34 also increased the proliferation of murine macrophages and breast cancer cells. Experiments on the protein level demonstrated the activation of MAPK/ERK in murine macrophages and breast cancer cells by recombinant IL-34. These preliminary data indicate that TAM-breast cancer cell interactions induce IL-34 production associated with induction of proliferation and the migratory capacity of tumor and stromal cells, which together may influence the invasiveness of the developing tumor. Here we propose IL-34 as a target for the treatment of breast cancer which may affect the accumulation of TAMs and improve breast cancer prognosis.

Citation Format: Sandun Gunawardhana, Karin Zins, Trevor Lucas, Dietmar Abraham. Novel CSF-1 receptor ligand IL-34 modulates macrophage-breast cancer cell crosstalk. [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 1160. doi:10.1158/1538-7445.AM2014-1160
Abstract 1166: CXCR7-mediated signaling axis regulates breast cancer growth and metastasis by modulating tumor microenvironment
Authors: Wani Nissar A.
Published: May 2015

Chemokine CXCL12 has been shown to bind chemokine receptor CXCR4 and CXCR7. Although extensive studies have been done to elucidate the role of CXCL12/CXCR4 signalling axis in tumor growth and metastasis, not much is known about the role of CXCL12/CXCR7 signalling axis in regulating breast cancer growth and metastasis. In order to study the role of CXCR7 in breast cancer growth and metastasis, we employed two models-i) 4T1 and CXCR7 downregulated 4T1 (4T1 shRNA-CXCR7) and ii) highly metastatic 4T1.2. First we observed CXCL12 induced and CXCR7 dependent migration of breast cancer cells and CXCL12 induced activation of ERK, STAT3 and cyclin D1, a downstream target of STAT3. In orthotopic syngenic models of breast cancer, we found reduced tumor growth and metastasis of breast cancer cells genetically silenced or pharmacologically inhibited for CXCR7 or its downstream target STAT3. The downregulation or inhibition of CXCR7 or its downstream target reduced tumor proliferation, angiogenesis and macrophage (M2) recruitment to the tumor site. Further analysis revealed the role of CXCR7 in secretion of cytokines (GMCSF, M-CSF and MCP-2), and expression of MMP-9, MMP-2 and VCAM-1. Moreover, we observed that CXCR7 is highly expressed in breast cancer tissues and positively correlates with disease progression from invasive ductal carcinoma (IDC) to metastasis. These observations highlight the importance of the CXCR7 axis in regulating breast cancer growth and metastasis. Blocking the CXCL12/CXCR7/STAT3 axis and macrophage recruitment to the tumor site with CXCR7 or STAT3 inhibitor may lead to the development of novel therapeutic strategies against breast cancer.

Citation Format: Nissar A. Wani. CXCR7-mediated signaling axis regulates breast cancer growth and metastasis by modulating tumor microenvironment. [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 1166. doi:10.1158/1538-7445.AM2014-1166
Abstract 119: Overexpression SOCS-1 may inhibit invasion and metastasis of cervical cancer
Authors: Kim Moon-Hong , Kim Wonwoo , Kang Miae , Seol Hyesil , Jeong Jae-Hoon
Published: May 2015

Purpose: To investigate whether the overexpression of SOCS-1 affects radiosensitivity, invasion, and metastasis in xenograft nude mouse model.

Methods: We established stable SOCS-1-overexpressing ME-180 cell line with pLNCX2-mSOCS1-Myc. Also we established xenograft nude mouse model for tumor growth delay experiment. Immunohistochemistry was employed to elucidate the pattern of expression of VEGF, CD-31, MMP-2, and TUNEL assay for apoptosis in SOCS-1 overexpressing ME-180 cells. Tumor cells were injected to right thigh of mouse to form cancer mass. Lung metastasis assay was performed in day 45 and 60. Metastatic nodules in both lungs were obtained and counted for statistical analysis.

Results: Tumor growth delay after 5 Gy radiation was observed in SOCS overexpressing tumor compared to wild type tumor but the difference did not show the statistical significance (p>0.05, Mann-Whitney U test). Immunohistochemistry showed that decreased MMP-2 expression, decreased VEGF expression, decreased CD-31 expression, and significantly elevated index of apoptotic body in SOCS-1 overexpressing tumor. Lung metastasis assay showed statistically significant decrement of metastatic nodule count in SOCS-1 overexpressing tumor (p<0.05, Mann-Whitney U test).

Conclusion: Decrement of radiosensitivity was statistically insignificant in SOCS-1 overexpressing tumor while invasion, angiogenesis, and metastasis were inhibited. Also apoptosis was increased in SOCS-1 overexpressing tumor.

Citation Format: Moon-Hong Kim, Wonwoo Kim, Miae Kang, Hyesil Seol, Jae-Hoon Jeong. Overexpression SOCS-1 may inhibit invasion and metastasis of cervical 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 119. doi:10.1158/1538-7445.AM2014-119
Abstract 1220: Investigation of the biological properties of human breast cancer in a nude rat model
Authors: Mokhtari Reza Bayat , Nofiele Joris Tchouala , Islam Syed S. , Yeger Herman , Cheng Hai-Ling Margaret
Published: May 2015

Developing imaging technology for cancer diagnosis and treatment monitoring is best performed on larger pre-clinical animal models recapitulating tumor growth and metastasis more similar to that found in humans. Such models are amenable to morphological and functional imaging techniques at spatial resolutions appropriate for animal imaging and, translatable to human imaging. In comparison to mouse xenografts, rat models offer the advantages of clinical imaging capabilities, development of xenograft and metastatic models, in orthotopic sites. Human breast cancer cell lines MDA-MB-231 and MCF-7 show a variable take rate in mice, ∼ 68% incurring considerable cost and time. Having a more reproducible method is needed for breast cancer studies. Here we present evidence for development of a novel method in the immune-compromised nude rats. Spheroids grown under stem cell conditions were derived from human breast adenocarcinoma estrogen dependent (MCF-7, ZR-75-1) and independent (MDA-MB-231) lines, and xenografted in both subcutaneous and orthotopic (fat pad) sites in the nude rat. To verify that rat tumors could be studied in detail at spatial resolutions achievable on a clinical 3 Tesla scanner, high-resolution magnetic resonance imaging was performed to identify vascular, viable, and necrotic tumor regions. The breast cancer phenotype and was confirmed by immunohistochemistry for ER and HER2. Tumors were characterized with proliferation marker Ki67, vascularization by CD34 and VEGF, and presence of hypoxic regions by HIF1α. Our results indicated that spheroids from all three lines readily generated tumors independent of exogenous estrogen. MRI is an effective and sensitive method for investigating the biological behavior and vascularization of breast cancer. These findings offer a potential novel method for pre-clinical study and investigation of human breast cancer.

Citation Format: Reza Bayat Mokhtari, Joris Tchouala Nofiele, Syed S. Islam, Herman Yeger, Hai-Ling Margaret Cheng. Investigation of the biological properties of human breast cancer in a nude rat model. [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 1220. doi:10.1158/1538-7445.AM2014-1220
Abstract 1221: Establishment and characterization of a chemoresistant glioblastoma cell line from an Iraqi patient
Authors: Al-Shammari Ahmed M. , Al-Juboory Ahmed A. , Al-Mukhtar Asmaa A. , Ali Amal M. , Al-Hili Zaid A. , Yaseen Nahi Y.
Published: May 2015

Glioblastoma multiforme is the most common and aggressive malignant primary brain tumour in humans, and has a very poor prognosis. As it is a well known fact that environmental factors especially radiation exposure is a major risk for glioblastoma multiforme and Iraq had been exposed in a relatively short period to 2 international wars with documented radiation exposure, so this paper might give justification for establishment of a new glioblastoma cell line. This cell line has been established from a cerebral glioblastoma multiforme in a 72-year-old Iraqi man who underwent surgery for an intracranial tumour in 2005. The morphology, growth kinetics, karyotype, immunocytochemistry and angiogenesis factor expression profile of this ANGM5 cell line were studied. The cell line has been grown continuously for more than 200 serial passages in standard culture media presenting no sign of cell senescence. The cultured cells are elongated or multipolar in shape and the population doubling time is 28 hours. The karyotype is complex and the chromosomal number varied between 46 and 48. The cell line is resistant to Temozolomide, cisplatin, vincristine and etoposide. This chemoresistance could be explained by the overexpression of the breast cancer resistance protein (BCRP) which found to be overexpressed by this cell line. Immunocytochemistry also showed that this cell line exhibited several characteristic features of glioblastomas, analysis of glial markers demonstrated that cells are positive for glial fibrillary acidic protein (GFAP), and negative for nestin and calbindin. Protein microarray analysis showed very high production of TIMP2 as well as other factors that are important for the invasiveness and aggressiveness of glioblastoma. This glioblastoma cell line is a useful addition to the cell lines currently available for study of the pathobiology and chemoresistance properties of glioblastoma multiforme as well as for antitumour drug discovery.

Citation Format: Ahmed M. Al-Shammari, Ahmed A. Al-Juboory, Asmaa A. Al-Mukhtar, Amal M. Ali, Zaid A. Al-Hili, Nahi Y. Yaseen. Establishment and characterization of a chemoresistant glioblastoma cell line from an Iraqi patient. [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 1221. doi:10.1158/1538-7445.AM2014-1221
Abstract 1224: A patient derived xenograft (PDX) platform for development of next generation KIT kinase inhibitors in imatinib-resistant gastrointestinal stromal tumors (GIST)
Authors: Mullins Chelsea , Ricono Jill , Carson Patrick , Habets Gaston , Shellooe Rafe , Nguyen Hoa , Broudy Thomas , Mirsaidi Cyrus , Nair Praveen
Published: May 2015

Despite best efforts in the treatment of gastrointestinal stromal tumors (GIST), patients continue to face a poor prognosis. That said, introduction of imatinib into clinical practice has vastly improved outcomes for KIT positive patients. However, as with many kinases, resistance has developed into a clinical dilemma, rendering the drug ineffective. A number of resistance mutations have previously been identified including the well known D816 activating mutation. To overcome this challenge, we have established a set of GIST patient derived xenograft models to recapitulate the original patient tumor, including genetics of resistance. In this work, we show four primary GIST samples which were selected based on mutational and clinical profile. PDX models were developed in immunocompromised mice and were further characterized by immunohistochemistry, additional sequencing and pharmacological efficacy. We then evaluated the efficacy of chemotherapeutic agents in these models. We describe imatinib resistance mutations, and demonstrate in vivo efficacy of dasatinib over imatinib in the resistant GIST PDX model. Take together, this data validates these GIST PDX models as a novel platform for the evaluation of new drug candidates to better delay and circumvent resistance now found in the clinic.

Citation Format: Chelsea Mullins, Jill Ricono, Patrick Carson, Gaston Habets, Rafe Shellooe, Hoa Nguyen, Thomas Broudy, Cyrus Mirsaidi, Praveen Nair. A patient derived xenograft (PDX) platform for development of next generation KIT kinase inhibitors in imatinib-resistant gastrointestinal stromal tumors (GIST). [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 1224. doi:10.1158/1538-7445.AM2014-1224
Abstract 1235: Targeting the DNA binding domain of the androgen receptor as a potential therapy for prostate cancer
Authors: Dalal Kush , Li Huifang , Moniri Mani R. , Ban Fuqiang , Sharma Aishwariya , Cherkasov Artem , Rennie Paul S.
Published: May 2015

The Androgen Receptor (AR) is a ligand inducible transcription factor that drives expression of genes contributing to the growth, recurrence and metastasis of prostate cancer (PCa). The AR is activated by steroid hormones (androgens) that bind to a C-terminal ligand-binding domain (LBD) and cause the transcription factor to enter the nucleus, dimerize and interact with DNA via its conserved DNA-binding domain (DBD). Accordingly, treatment for PCa involves reducing DHT production or using small molecules (i.e. antiandrogens) to compete with the specific interaction between hormones and the AR-LBD, preventing AR activation. This treatment will eventually fail as recurrent PCa changes into a castration resistant form (CRPC) where AR signalling continues even in the absence of androgens. The occurrence of CRPC is driven, in part, by mutations in the AR-LBD that alter the binding of antiandrogens, or by the presence of constitutively active AR splice variants that lack the LBD, effectively removing the site of action of conventional antiandrogens.

Here, we describe a surface exposed pocket on the AR-DBD, based on its 3D-crystal structure, which may qualify as an alternative target for small molecule inhibitors of AR transcriptional activity. Following virtual screening of many compounds, we characterize the interaction of the AR-DBD with various potential inhibitors and measure the efficacy of these agents to block the transcriptional activity of both full-length and splice variant AR in cell culture. Drug-inhibition was reduced upon site-directed mutagenesis of amino-acid residues in the DBD that are suspected of interacting with the compounds. Biochemical approaches are used to further examine the physical interaction of the purified AR-DBD protein with small molecules and to investigate their effect on protein-DNA interactions in vitro. Lastly, we use confocal microscopy of live PCa cells to investigate the effect of our compounds on the spatial localization and dimerization of fluorescently tagged AR proteins. These results illustrate the feasibility of targeting the AR-DBD to potentially inhibit the reactivation of AR transcriptional activity that is observed in CRPC.

Citation Format: Kush Dalal, Huifang Li, Mani R. Moniri, Fuqiang Ban, Aishwariya Sharma, Artem Cherkasov, Paul S. Rennie. Targeting the DNA binding domain of the androgen receptor as a potential therapy for prostate 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 1235. doi:10.1158/1538-7445.AM2014-1235
Abstract 124: Effect of RKIP on MMP expression and regulation of breast cancer metastasis
Authors: Datar Ila J. , Feng Jingwei , Ren Gang , Li Zehui , Qiu Xiaoliang , Al-Mulla Fahd , Bitar Milad S. , Yeung Miranda , Yeung Kam
Published: May 2015

Raf Kinase Inhibitory Protein or RKIP was initially identified as a Raf-1 binding protein using the yeast 2-hybrid screen. RKIP inhibits the activation phosphorylation of MEK by Raf-1 by competitively inhibiting the binding of MEK to Raf-1 and thus exerting an inhibitory effect on the Raf-MEK-ERK pathway. RKIP has been identified as a metastasis suppressor gene. Metastasis is a complex multiple-step process. Expression of RKIP is low in cancer metastases. Although primary tumor growth was unaffected, re-expression of RKIP inhibits cancer metastasis. Mechanistically, RKIP constrains metastasis by inhibiting angiogenesis, local invasion, intravasation, and colonization. The molecular mechanism of how RKIP inhibits these individual steps remains undefined. Using an unbiased PCR based screening and by analyzing DNA microarray expression datasets we observed that the expression of multiple metalloproteases (MMP) including MMP1, MMP3, MMP10 and MMP13 were negatively correlated with RKIP expression in breast cancer cell lines and clinical samples. Since expression of MMPs by cancer cells is important for cancer metastasis, we hypothesize that RKIP may mediate suppression of breast cancer metastasis by inhibiting multiple MMPs. In this study we show that the expression signature of RKIP and MMPs is better at predicting high metastatic risk than the individual gene. Using a combination of loss- and gain-of-function approaches, we found that MMP13 is the cause of RKIP-mediated inhibition of local cancer invasion. Interestingly expression of MMP13 alone was not sufficient to reverse the inhibition of breast cancer cells metastasis to the lung due to the expression of RKIP. We found that RKIP negatively regulated MMP13 through the Erk2-Fra1 transcriptional axis and the repression of MMP13 by RKIP requires the presence of the a TPA-responsive element in the MMP13 proximal promoter. Together, our findings indicate that RKIP inhibits cancer cell invasion, in part, via MMP13 inhibition. These data also implicate RKIP in the regulation of MMPs transcription, suggesting a potential mechanism by which RKIP inhibits tumor progression and metastasis.

Citation Format: Ila J. Datar, Jingwei Feng, Gang Ren, Zehui Li, Xiaoliang Qiu, Fahd Al-Mulla, Milad S. Bitar, Miranda Yeung, Kam Yeung. Effect of RKIP on MMP expression and regulation of breast cancer 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 124. doi:10.1158/1538-7445.AM2014-124
Abstract 1241: Kaempferol suppresses solar ultraviolet radiation-induced skin cancers by targeting RSK2 and MSK1
Authors: Yao Ke , Chen Hanyong , Lee Mee-Hyun , Langfald Alyssa , Kim Myoung Ok , Yu Dong Hoon , Liu Kangdong , Ma Wei-Ya , Bode Ann M. , Dong Ziming , Dong Zigang
Published: May 2015

Ultraviolet (UV) irradiation is the leading factor in the development of skin cancer, which is the most common form of cancer in the United States. Discovering novel chemopreventive agents against this disease is extremely important. Kaempferol, a natural flavonol isolated from tea, broccoli, grapes, apples and other plant sources, is known to have anticancer activity, but its molecular mechanisms and direct target(s) in cancer chemoprevention are still unclear. In this study, our pull-down assay results showed that RSK2 and MSK1 directly interact with kaempferol in both ex vivo and in vitro systems. ATP competition and in vitro kinase assay data revealed that kaempferol interacts with RSK2 and MSK1 at the ATP-binding pocket and inhibits their respective kinase activities. Mechanistic investigations determined that kaempferol acts as an inhibitor of RSK2 and MSK1 kinase activities to attenuate solar UV-induced phosphorylation in mitogen-activated protein kinase signaling cascades in JB6 P+ mouse skin epidermal cells. In a mouse skin tumorigenesis study, kaempferol significantly suppressed solar UV-induced skin carcinogenesis. Further analysis showed that the kaempferol-treated group had a substantial reduction in solar UV-induced phosphorylation of CREB and c-Fos in mouse skin. Taken together, our results identify kaempferol as a safe and novel chemopreventive agent against solar UV-induced skin carcinogenesis that acts by targeting RSK2 and MSK1.

Citation Format: Ke Yao, Hanyong Chen, Mee-Hyun Lee, Alyssa Langfald, Myoung Ok Kim, Dong Hoon Yu, Kangdong Liu, Wei-Ya Ma, Ann M. Bode, Ziming Dong, Zigang Dong. Kaempferol suppresses solar ultraviolet radiation-induced skin cancers by targeting RSK2 and MSK1. [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 1241. doi:10.1158/1538-7445.AM2014-1241
Abstract 1261: Targeting IL-11Rα inhibits osteosarcoma pulmonary metastasis in an orthotopic xenograft mouse model
Authors: Lewis Valerae O. , Devarajan Eswaran , Hughes Dennis PM
Published: May 2015

Osteosarcoma is the most common primary tumor of bones. In the past three decades treatment paradigms and survival rates have not improved. While osteosarcoma clinical phenotype has been well characterized, the underlying tumor molecular biology remains unclear. We have previously demonstrated that IL-11Rα is overexpressed in primary and metastatic osteosarcoma. Recently investigators identified that IL-11 and IL-11Rα play a role in the development of metastasis in breast and gastrointestinal cancer. However the role of IL-11/IL-11Rα signaling in osteosarcoma is unknown. Using in vitro and in vivo osteosarcoma models we investigated the role of IL-11/IL-11Rα in osteosarcoma metastasis. The purpose of our study is to determine the effect of the knockdown of IL-11Rα in osteosarcoma. We stably knocked down IL-11Rα expression in a highly metastatic osteosarcoma cell line, using a lentiviral short hair-pin (shRNA) based approach. Knockdown of IL-11Rα led to inhibition of STAT-3(Tyr705) protein phosphorylation, cell growth, migration, invasion and colony formation in vitro. To determine the role of IL-11Rα in osteosarcoma tumorigeniesis and metastasis, we used an orthotopic metastatic model. In this model, mice injected with KRIB-IL-11Rα shRNA cells developed significantly less metastasis than control mice injected with KRIB control shRNA (P<0.01). The relevance of our data to human conditions was studied by immunohistochemical staining of IL-11Rα protein in human osteosarcoma biopsy samples containing primary tumor and lesions, significant difference (p<0.01) in expression distribution between primary and metastatic osteosarcoma lesions and a trend toward an impact on survival was noted. In conclusion, we showed that IL-11Rα plays an important role in the development of metastasis in osteosarcoma and blockade of this axis may inhibit the formation of metastasis and may provide an excellent target for the treatment of osteosarcoma.

Citation Format: Valerae O. Lewis, Eswaran Devarajan, Dennis PM Hughes. Targeting IL-11Rα inhibits osteosarcoma pulmonary metastasis in an orthotopic xenograft mouse model. [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 1261. doi:10.1158/1538-7445.AM2014-1261
Abstract 129: AIP1 suppresses tumor metastasis by regulating tumor microenvironment and metastatic niche
Authors: Min Wang
Published: May 2015

AIP1 (also called DAB2IP, a new member of RAS-GAP family protein) is downregulated in a variety of human cancer tissues. We and others have recently reported that AIP1 functions as a tumor suppressor in prostate cancer models. However, the role of AIP1 in regulating tumor microenvironment and metastatic niche is not known. In the present study, we implanted tumor cells with the same genetic background into C57BL/6 wild type (WT) and AIP1-deficient (AIP1-KO) mice to address this question. In subcutaneous (SQ) allograft models using B16 melanoma and LLC Lewis lung carcinoma, tumor growth and metastasis to multiple organs including lymph node, lung, liver, spleen and bone marrow were significantly enhanced in AIP1-KO host, suggesting AIP1 in tumor microenvironment limits tumor metastases. However, a direct intravenously injection of tumor cells resulted in no differences in tumor metastasis between WT and AIP1-KO mice, indicating AIP1 in host regulates tumor metastasis at steps prior to extravasation to target tissues. We examined effects of AIP1 deletion on the induction of local inflammation, tumor cell epithelial-mesenchymal transition (EMT), generation of metastatic routes (formation of new blood and lymphatic vessels) and establishment of pre-metastatic niche in distance tissues. SQ tumors in AIP1-KO host showed more profound increases in macrophage infiltration, tumor cell EMT, intratumoral angiogenesis and peri-tumoral lymphangiogenesis as well as lymphangiogenesis in lymph nodes prior to the detection of metastatic tumor cells. Finally, SQ tumor induced an establishment of pre-metastatic niche in lung as indicated by increased immunostaining for fibronectin and accumulation of VEGFR1, c-Kit and CXCR-positive cells which were augmented in AIP1-KO mice. Importantly, WT recipient with AIP1-KO bone marrow-derived cells showed similar phenotype to AIP1-KO mice in tumor growth and metastasis. Taken together, our data suggest that AIP1 functions as tumor metastasis suppressor by regulating tumor microenvironment, metastatic routes and pre-metastatic niche.

Citation Format: Wang Min. AIP1 suppresses tumor metastasis by regulating tumor microenvironment and metastatic niche. [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 129. doi:10.1158/1538-7445.AM2014-129
Abstract 1293: Familial melanoma by histology and age guiding clinical genetic counseling: joint data from five Nordic countries
Authors: Fallah Mahdi , Pukkala Eero , Sundquist Kristina , Tretli Steinar , Olsen Jørgen H. , Tryggvadottir Laufey
Published: May 2015

Background: We aimed to estimate lifetime cumulative risk of melanoma (CRM) in relatives of patients with melanoma by histology and age at diagnosis in patients and relatives.

Methods: A population-based cohort of 238,724 first-degree relatives of 46,091 melanoma patients diagnosed in 1955-2010 in Nordic countries was followed for cancer incidence.

Findings: The CRM (0-79 years) in first-degree relatives of a patient with superficial spreading (SSM), nodular (NM), or lentigo maligna (LMM) melanoma was quite similar, ranging from 2.5% to 2.9%, which represents about 2-fold increase over the general population risk. When one melanoma patient in the family was diagnosed before age 30, the CRM was 3.2%. When there were ≥2 melanoma patients diagnosed before age 30 in a family, CRM for relatives was 13.8%, 5.8% for diagnoses at age 30-59, and 4.8% for diagnoses at age 60 or older. Depending on age at diagnosis of same-sex twins (not known whether monozygotic or dizygotic), their CRM was 7.4-21.3%. Although no familial case of concordant histological types of acral lentiginous/desmoplastic/compound nevus/spindle cell melanomas or malignant blue nevus (MBN) was found, familial risks of discordant histological types of melanoma were interchangeably high for most of the types, e.g. higher risk of SSM when a first-degree relative had NM [standardized incidence ratios (SIR)=2.6, 95%CI=2.1-3.3, n=72] or ALM (4.0, 95%CI=1.5-8.8, n=6) and vice versa. There was a tendency toward concordant age at diagnosis of melanoma among relatives of melanoma patients.

Interpretation: This study provides clinically relevant absolute and relative risk estimates for family members of melanoma patients by age at diagnosis in patients and relatives. Our finding may suggest that the role of genetic or environmental factors in the differentiation of histological types of melanoma is not as strong as its role in the development of melanoma as such.

Citation Format: Mahdi Fallah, Eero Pukkala, Kristina Sundquist, Steinar Tretli, Jørgen H. Olsen, Laufey Tryggvadottir. Familial melanoma by histology and age guiding clinical genetic counseling: joint data from five Nordic countries. [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 1293. doi:10.1158/1538-7445.AM2014-1293
Abstract 1313: Anti-tumorigenic effects of phenformin in human endometrial cancer cells
Authors: Jackson Amanda L. , Kilgore Joshua E. , Qiu Haifeng , Zhou Chunxiao , Gehrig Paola A. , Bae-Jump Victoria L.
Published: May 2015

Introduction: The anti-diabetic drugs, metformin and phenformin, are thought to have anti-tumorigenic benefits. Preclinical studies suggest that phenformin may be more potent for inhibiting tumor growth than metformin. Our objective was to assess the anti-tumorigenic effects of phenformin in endometrial cancer (EC) cell lines and primary cultures.

Methods: The EC cell lines, ECC-1 and Ishikawa, were treated with phenformin under low glucose (2 mM), normal glucose (5.5 mM) and high glucose (25 mM) conditions. EC tumors were collected from consenting patients, at the time of surgical staging for primary culture. Cell proliferation was assessed by MTT assay. Apoptosis was analyzed by Annexin V-FITC assay. Cellometer evaluated cell cycle progression. Invasion was demonstrated by transwell invasion assay. Adhesion was assessed by ELISA. Western blotting was performed to determine expression of the downstream targets of phenformin.

Results: Phenformin inhibited proliferation in a dose-dependent manner in both EC cell lines, within 48-72 hrs of exposure (IC50=1 mM, p=0.0004-0.009). Inhibition of proliferation was seen for all glucose concentrations, but greater potency was observed under low glucose conditions. Treatment with phenformin resulted in G1 arrest, induction of apoptosis (p=0.022-0.017) and inhibition of adhesion (p=0.0053-0.0144) and invasion (p=0.0063-0.05). Phenformin increased p21 and p27 and decreased cyclin D, cyclin E2, CDK4 and CDK 6, corresponding with G1 arrest. Phenformin significantly inhibited proliferation in 62% (8/13) of the EC primary cultures, within 48-72 hrs of exposure (IC50 0.1-5mM, p=0.00001-0.016). 3 additional EC samples (23%) were significantly inhibited by phenformin but did not reach an IC50 (p=0.0003-0.018), and 2/13 (15%) had no response to phenformin. Western blot analysis demonstrated that phenformin increased phosphorylation of AMPK and decreased phosphorylation of S6 coincident with inhibition of proliferation in the EC cell lines and primary cultures.

Conclusion: Phenformin potently inhibited cell growth via G1 arrest and induced apoptosis in EC cell lines and the majority of primary cultures. Inhibition of adhesion/invasion was also seen with phenformin. Continued work is needed to explore whether phenformin has superior benefits over metformin.

Citation Format: Amanda L. Jackson, Joshua E. Kilgore, Haifeng Qiu, Chunxiao Zhou, Paola A. Gehrig, Victoria L. Bae-Jump. Anti-tumorigenic effects of phenformin in human endometrial 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 1313. doi:10.1158/1538-7445.AM2014-1313
Abstract 1359: Identification of a novel hypermethylated tumor suppressor gene in human bladder cancer
Authors: Yeh Chia-Ming , Chen Pi-Che , Huang Wen-Yu , Shen Cheng-Huang , Hsu Cheng-Da , Chan Michael W.
Published: May 2015

Bladder cancer is the sixth most common cancer in the world and the incidence is particularly high in southwestern Taiwan. Although bladder cancer patients have a low mortality rate, long term follow-up with repeated cystoscopy is required due to the high recurrence nature of the tumor. Therefore, a non-invasive detection assay is urgently required for bladder cancer patients. Aberrant promoter hypermethylation which is considered as a hallmark of cancer for over a decade, plays an important role in controlling cancer progression. Nevertheless, detection of promoter hypermethylation in bodily fluid has been implicated as a non-invasive and sensitive tool for cancer diagnosis. We aim to identify novel markers that are hypermethylated in bladder cancer. In this regard, by using illumina 27K CpG island methylation array, we identified several novel targets that are hypermethylated in 7 bladder cancer patient samples but not in primary normal human urothelim. Combined bisulfite restriction analysis (COBRA) and bisulphite pyrosequencing confirmed the array result in bladder cancer cell lines. Further, bisulphite pyrosequencing demonstrated that a significantly higher methylation level of those targets in cancer samples with higher histological grade (P < 0.05) and pathological stage (P < 0.05). The potential of non-invasive diagnosis of bladder cancer detection by using the methylation of these targets in voided urine samples is under-way. The tumor suppressive function of these methylated targets in bladder cancer deserves further investigation.

Citation Format: Chia-Ming Yeh, Pi-Che Chen, Wen-Yu Huang, Cheng-Huang Shen, Cheng-Da Hsu, Michael Wy Chan. Identification of a novel hypermethylated tumor suppressor gene in human bladder 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 1359. doi:10.1158/1538-7445.AM2014-1359
Abstract 1366: Discovery of biomarkers for antitumor drug resistance using 450K methylation data of NCI60
Authors: Nogales Vanesa , Moutinho Catia , Martinez-Cardús Anna , Varma Sudhir , Killian J. Keith , Reinhold William C. , Meltzer Paul S. , Pommier Yves , Esteller Manel
Published: May 2015

The National Cancer Institute Antitumor Cell line panel NCI60 is a very frequently used collection of cancer cell lines that has been largely characterized in different biological aspects. Data from proteomic and RNA expression analyses of this panel is already available to the scientific community. Also, the IC50 data for more than 100,000 different compound treatments is accessible in the developmental therapeutic program of NCI (dtp). However, very little has been explored about the epigenetic component of this panel. We have performed a thorough correlational study combining the available molecular information of NCI60, the data of our 450K NCI60 dataset and the IC50 data from the dtp to discover candidate genes for new biomarkers of resistance to antitumor agents. Finding new biomarkers to determine the most probable response to the therapy seems to be a key stage in the development of personalized medicine. Due to its stability and how easy it is to detect its level of methylation, DNA methylation changes appear to be a perfect candidate mark for the construction of this new generation of biomarkers. In this study we obtained multiples candidate genes whose methylation correlated to resistance to different drugs. In order to perform validation analyses we chose a gene that showed a strong positive correlation with the resistance to several DNA damage agents (DDA).Using overexpression and silencing assays we were able to demonstrate that the expression of this gene has a profound influence on the cell sensitivity to the DDA. Furthermore, through gene expression validation assays, such as 5-azacytidine treatments, we demonstrated that the promotor methylation of this gene is associated to its non-expression. In addition, we observed that this gene methylation correlates with a lower overall survival in an ovarian cancer cohort treated with platinum agents. Little is known about our candidate gene but it seems to be related to the DNA repair machine and also to have a function in the cell cycle control. These features could give an explanation to the importance of its presence in the response to DDA treatment. Our studies show that the analysis of DNA methylation changes in cancer cells in combination with other molecular alterations can be used in correlational studies to find trustworthy biomarkers for antitumor drug resistance.

Citation Format: Vanesa Nogales, Catia Moutinho, Anna Martinez-Cardús, Sudhir Varma, J. Keith Killian, William C. Reinhold, Paul S. Meltzer, Yves Pommier, Manel Esteller. Discovery of biomarkers for antitumor drug resistance using 450K methylation data of NCI60. [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 1366. doi:10.1158/1538-7445.AM2014-1366
Abstract 1400: Extended concurrent gene signatures of ER, HER2 and disease-free survival in breast cancers
Authors: Huang Chi-Cheng , Tu Shih-Hsin , Huang Ching-Shui , Lien Heng-Hui , Lai Liang-Chuan , Chuang Eric Y.
Published: May 2015

Our recent work had identified concurrent gene signatures which highlighted the interplay between copy number variation (CNV) and differential gene expression (GE) for Han Chinese breast cancers. The merit of the approach is the discovery of biomarkers not readily identified by conventional GE only data, for which phenotype-correlation or gene variability is the criteria of gene selection.

A total of 31 array CGH and 83 GE arrays (29 samples with data from both platforms) were performed. Potential targets of cancer therapy were revealed by Genomic Identification of Significant Targets in Cancer (GISTIC) from 31 array CGH. We used these concurrent genes as well as genes with significant GISTIC scores to derive signatures associated with clinical ER, HER2 status, and disease-free survival.

The most common gain regions were 1q21.2 and 17q12-q21.1 while the most common lost region was 22q11.23. In subgroup analysis, recurrent gains for ER positive cancers were 8p23.1, and 8p11.23-11.21. For HER2 overexpressing tumors, recurrent gain was 17q12-21.1 and recurrent losses were 8p11.22, 8p12, and 8p23.1. We identified 1,584 concurrent genes from 29 samples assayed for both array CGH and GE microarrays, and enriched concurrent gene sets for ER, HER2 and disease-free survival were identified independently from our 83 GE arrays (GSE48391) and two series with Han Chinese origin (GSE5460 and GSE20685) as well as three studies of Western countries (GSE7390, GSE2034 and GSE3494). Signature genes were consensus genes from leading edge analysis across all studies, and we used supervised partial least square (PLS) regression to derived predictive model of clinical ER, HER2 and disease-free survival with each signature gene further weighted by the coefficient of concurrence as an additive tuning parameter. The predictive accuracy was 92.3% for ER and 84.6% for HER2 in 208 Han Chinese breast cancers and was 87.3% for ER in 735 breast cancers in Western countries. For five studies with disease-free survival data (no survival data for GSE5460), prognostic discrepancy was observed in four studies except GSE3493 between high-risk and low-risk patients predicted with the concurrent signature.

We conclude that parallel analysis of CGH and microarray data, in conjunction with known gene expression patterns, can be used to identify biomarkers with prognostic values in breast cancer.

Citation Format: Chi-Cheng Huang, Shih-Hsin Tu, Ching-Shui Huang, Heng-Hui Lien, Liang-Chuan Lai, Eric Y. Chuang. Extended concurrent gene signatures of ER, HER2 and disease-free survival in breast cancers. [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 1400. doi:10.1158/1538-7445.AM2014-1400