bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 224: Critical variables for automated protein immunodetection using western blot
Authors: Margulis Alexander , Chimento David P. , Yukhananov Russ 
Published: July 2017

Abstract
Reliable immunodetection of proteins is a critical step in the discovery of novel biomarkers. It is often performed manually. The immunodetection protocol consists of multiple steps including blocking of nonspecific binding sites, incubation using primary and secondary antibodies and extensive washing between steps that easily introduce bias and errors. The quality of results depends on multiple subjective and objective factors such as the qualification and technical skills of the personnel performing the assay and the accuracy of temporal and temperature control, especially during the immunodetection step. BlotCycler™, automated western blot processor, use fluidic control system that allows to eliminate the variability associated with immunodetection and to achieve higher sensitivity by optimized washing procedure. We used BlotCycler to analyze critical variables for immunodetection including temperature, incubation time and reagent mixing. Relatively small changes in duration and temperature of incubation significantly affect not only the intensity of signal but also the specificity of antibody interaction with antigen. Automated immunodetection at 4ºC significantly improve the specificity of detection and sensitivity that allows reproducible detection of low expressing proteins. Automated processing using BlotCycler allows optimization and control of critical variables leading to improved reproducibility and specificity of immunodetection and should be routinely used for biomarker candidate discovery and confirmation, reliable protein quantification, primary antibody specificity testing, and optimization of primary and secondary antibodies concentrations.
Citation Format: Alexander Margulis, David P. Chimento, Russ Yukhananov. Critical variables for automated protein immunodetection using western blot [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 224. doi:10.1158/1538-7445.AM2017-224
Abstract 2240: “Smart” selenium compound ISC-4 inhibits androgen receptor signaling and induces ROS- and p53-dependent apoptosis in LNCaP prostate cancer cells
Authors: Wu Wei , Pramanik Kartick , Jiang Cheng , Amin Shantu , Sharma Arun K. , Lu Junxuan 
Published: July 2017

Abstract
“Smart” selenium compound phenylbutyl isoselenocyanate (ISC-4) was developed by substituting sulfur in cancer chemopreventive compound phenylbutyl isothiocyanate (PBITC) with selenium, resulting in increased anti-cancer activity in several melanoma and colon cancer cell lines. The in vivo anticancer beneficial effects of ISC-4 have been reported in both melanoma and colon cancer mouse models as well. To explore the potential chemopreventive attributes of ISC-4 against prostate carcinogenesis, we selected LNCaP cells (with wild type p53 and functional androgen receptor (AR) signaling) to examine the growth suppression and apoptosis responses in cell culture and interrogated signaling mechanisms through pharmacological and siRNA knockdown approaches. Results show that when compared with PBITC, ISC-4 was more potent at suppressing cell growth and inducing apoptotic cell death in LNCaP cells. ISC-4 induced caspase-mediated apoptosis (as evidenced by cleavage of PARP) in dose- and time-dependent manners in association with decreased AR and its best known target prostate specific antigen (PSA) protein abundance and upregulation of p53-DNA damage response (DDR) protein expression levels: p53, its canonical target p21, and p-H2AX that marks DNA double-strand break sites. Similar to the rapid induction of reactive oxygen species (ROS) by isothiocyanates, ISC-4 exposure resulted in increased dihydroethidium-detectable ROS in a concentration-dependent manner. Preincubation with the ROS scavenger N-Acetyl-L-cysteine (NAC, 10 µM to 2 mM), markedly reduced ISC-4-induced cell death and attenuated p53, p21 and p-H2AX. Moreover, siRNA knockdown of p53 did not suppress ROS production, but decreased ISC-4-induced Annexin V-positive apoptosis, PARP cleavage and the DDR proteins. Taken together, these data suggest that ISC-4 inhibits LNCaP cell growth and survival with concomitant suppression of AR signaling and induction of ROS-p53-DDR mediated apoptosis. Given the pivotal significance of AR signaling in prostate epithelial survival, cancer genesis and progression, and the preservation of wild type p53 in precancerous lesions and early stage prostate cancer, our findings suggest attractive chemopreventive potential of ISC-4 against prostate carcinogenesis.
Citation Format: Wei Wu, Kartick Pramanik, Cheng Jiang, Shantu Amin, Arun K. Sharma, Junxuan Lu. “Smart” selenium compound ISC-4 inhibits androgen receptor signaling and induces ROS- and p53-dependent apoptosis in LNCaP prostate cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2240. doi:10.1158/1538-7445.AM2017-2240
Abstract 2242: Vitamin E δ-tocotrienol inhibits NF-κB activation by up-regulating A20 in macrophages and suppresses colitis-promoted colon tumorigenesis in mice
Authors: Yang Chao , Jiang Qing 
Published: July 2017

Abstract
Nuclear factor-κB (NF-κB) plays a key role in inflammatory responses and cancer development. Inhibition of NF-κB has been shown to attenuate inflammation and cancer progression. δ-Tocotrienol (δTE) is a natural vitamin E form rich in annatto seeds and has been shown to inhibit NF-κB in pancreatic cells. However, potential effect of δTE on NF-κB activation in immune cells and the underlying mechanism have not been investigated. In the present study, we found that δTE inhibited TNF-α induced activation of NF-κB, as indicated by diminished phosphorylation and degradation of IκBα in the cytosol, and phosphorylation of p65 in the nucleus. Furthermore, δTE potently inhibited TNF-α-induced phosphorylation of TAK1, a key upstream kinase that is required for the activation of NF-κB. Interestingly, δTE significantly increased the expression of A20 and CYLD, both of which are negative regulators of NF-κB. Knockout of A20 partially diminished δTE's anti-NF-κB effect, supporting the notion that induction of A20 plays a key role in δTE’s anti-NF-κB effect. To translate mechanistic findings to a whole body environment, we further investigated the effectiveness of δTE and its metabolite, δTE-13’-COOH, against azoxymethane (AOM) induced and colitis-promoted colon tumorigenesis in male BALB/c mice. We found that δTE-rich mixed tocotrienols (δTE /γTE ~8/1, at 0.035% diet) inhibited colon tumorigenesis, as indicated by 36% reduction of multiplicity of large adenomatous polyps (&gt;2mm(2), P=0.07) and decreased tumor surface area by 31% (P&lt;0.05). δTE-13’-COOH (at 0.04% diet) significantly suppressed multiplicity of total polyps by 25% (P=0.06) and large polyps by 54% (P&lt;0.01). Our study demonstrates that δTE has potent anti-NF-κB activity and this vitamin E form and its metabolite suppress colitis-promoted colon tumorigenesis in mice.
Citation Format: Chao Yang, Qing Jiang. Vitamin E δ-tocotrienol inhibits NF-κB activation by up-regulating A20 in macrophages and suppresses colitis-promoted colon tumorigenesis in mice [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2242. doi:10.1158/1538-7445.AM2017-2242
Abstract 2243: Elucidating the mechanism of action of molecular iodine on breast cancer cells
Authors: Xu Zack , Chen Xin , Yam Noymi , Chan kin , Nagavarapu Usha 
Published: July 2017

Abstract
Each year, over 1.7 million women are diagnosed with breast cancer worldwide and about 35% of these diagnosed are expected to die from the disease. A significant barrier to improving treatment of breast cancer lies in our incomplete understanding of breast cancer initiation and metastasis. Regular use of molecular iodine has been shown to reduce the sensitivity of breast cells to the proliferative effects of estrogen, resulting in normalization of breast tissue.The aim of the current study was to evaluate the mechanisms of molecular iodine on two common breast cancer sub-types using well-established breast cancer cell lines. In order to study Luminal A and triple-negative breast cancer in vitro, MCF7 (cells from a luminal A subtype) and MDA-MB231 (cells from a triple-negative subtype), were treated with molecular iodine at various concentrations to measure proliferation and cell death. This was subsequently followed by gene expression analysis of key important molecular markers, which are primarily responsible for cell growth and apoptosis. Primary human mammary epithelial cells derived from healthy female donor were used as an internal control. Data from this study indicated that molecular iodine had potent inhibitory effects on cell growth and showed a dramatic increase in cell death in both breast cancer cell lines. Gene expression analysis using quantitative RT-PCR further confirmed that cell cycle genes controlling G1-S phase transition were largely up-regulated. Changes were not seen in Cyclin B expression levels, which further suggest that cells were arrested before entry into cell division. BCL-2, PPAR-α and PPAR-γ were also up-regulated along with down-regulation of Caspase 3, suggesting molecular iodine induced cell death through activation of a caspase-independent apoptosis pathway. Interestingly, mesenchymal-epithelial transition (MET) occurrence was noticed upon molecular iodine treatment as indicated by sharp increase of GATA3 and E-Cadherin and significant down-regulation of Vimentin in invasive MDA-MB231 cells. Our current study enables us to understand a molecular mechanism controlling tumor cell growth and demonstrates potent cellular effects of molecular iodine on breast cancer cell lines. These results also demonstrate promising effects of molecular iodine for regulation of breast cancer EMT differentiation, which is required for tumor initiation and metastasis. Further studies are underway to determine possible effects of molecular iodine in these breast cancer subtypes using in vitro 3D models.
Note: This abstract was not presented at the meeting.
Citation Format: Zack Xu, Xin Chen, Noymi Yam, kin Chan, Usha Nagavarapu. Elucidating the mechanism of action of molecular iodine on breast cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2243. doi:10.1158/1538-7445.AM2017-2243
Abstract 2251: Tumor tissue gene expression in association with survival of triple-negative breast cancer
Authors: Wang Shuyang , Cai Qiuyin , Cai Hui , Bao Pingping , Wu Jie , Ye Fei , Zheng Wei , Zheng Ying , Shu Xiao-Ou 
Published: July 2017

Abstract
Triple-negative breast cancer (TNBC) is an aggressive type of cancer with limited treatment options. Previous studies have shown that gene expression profiles were associated with TNBC prognosis. Information on specific genes that are predictive of TNBC outcome is limited. Using data and samples from a cohort of 469 TNBC cases from Shanghai Breast Cancer Survival Study (SBCSS), we systematically evaluated expressions of 302 genes in tumor tissue with survival of TNBC. Genes included in the study are PAM50 genes, genes encoding drug metabolizing enzymes and, genes implicated in TNBC biology and progression based on literature reviews. Gene expression levels were measured in total RNA isolated from formalin-fixed paraffin-embedded breast cancer tissues using the NanoString nCounter assay. Cox regression was applied to evaluate disease-free survival (DFS) and overall survival (OS) in association with gene expression. Analysis was adjusted for known predictors of TNBC outcome, including age, disease stage, basal like subtype and DUSP4 gene expression. During a median follow-up of 5.3 years (range: 0.7-8.9 years), 100 deaths and 92 recurrences/breast cancer deaths were documented. Expression levels of 17 genes were significantly associated with OS, and 15 genes with DFS (P&lt;0.05). The top 5 most significant genes are EOMES (Hazard Ratio, HR = 0.88; 95% Confidence Interval, 95% CI: 0.82-0.96), GZMK (HR = 0.91, 95% CI: 0.85-0.97), TUBB3 (HR = 1.12, 95% CI: 1.03-1.21), SIT1 (HR = 0.90, 95% CI: 0.83-0.98) and ZNF224 (HR = 0.89, 95% CI: 0.81-0.97) for OS, and TUBB3 (HR = 1.14, 95% CI: 1.05-1.24), KRT14 (HR = 1.08, 95% CI: 1.02-1.14), BAG1 (HR = 1.19, 95% CI: 1.05-1.35), CD44 (HR = 0.73, 95% CI: 0.58-0.92) and EOMES (HR = 0.89, 95% CI: 0.82-0.97) for DFS. Study is on-going to replicate the findings of this study.
Citation Format: Shuyang Wang, Qiuyin Cai, Hui Cai, Pingping Bao, Jie Wu, Fei Ye, Wei Zheng, Ying Zheng, Xiao-Ou Shu. Tumor tissue gene expression in association with survival of triple-negative breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2251. doi:10.1158/1538-7445.AM2017-2251
Abstract 2253: 5 Gene prognostic signature for dedifferentiated liposarcomas
Authors: Ng Deanna , Shannon Nicholas , Ong Johnny , Teo Melissa 
Published: July 2017

Abstract
Introduction: Liposarcomas, despite being rare consist of a number of entities spanning low to high grade sarcomas. De-differentiated liposarcoma (DDLS) is associated with increased frequency of recurrence and lower overall survival. We aimed to identify gene expression changes associated with OS in DDLS. Secondary aims was to postulate a mechanism for DDLS.
Methods: We analysed data from gene expression omnibus (GEO, ID = GSE30929) to identify genes associated with DDLS (n=39). A signature representing genes overexpressed in DDLS was then applied to an independent dataset, utilising data from The Cancer Genome Atlas (TCGA, n=58 DDLS) to segregate samples based on gene expression and groupings compared for survival.
Results: A 5 gene signature was generated consisting of genes with increased expression in DDLS and its relation to OS. The signature was significantly enriched for genes involved in lipid metabolism (AQP7, PRKAR2B, FABP4 p&lt;0.01) and genes involved in the cell cycle and cell proliferation (FBN2, CEBPA p&lt;0.01). Segregation of DDLS in the TCGA cohort on the basis of median gene signature score gave a significant difference in overall survival. High score (stronger DDLS signature) was associated with a lower median survival(33.5 vs 76.4 months, p = 0.027), and poorer 2 year (63.4% vs 89.2%) and 5 year survival (20.6% vs 71.1%). The survival difference remained significant when applied to retroperitoneal DDLS only (n=51, median survival 34.5 vs 54.2 months, p = 0.034) and poorer 2 year survival (. A postulated mechanism is that DDLS have gained deficiency in AQP7, PRKAR2B and FABP4 leading to fatty acid and glycerol accumulation in cells, increasing cancer cell replication. CEBPA inhibits CDK4 and a deficiency of CEBPA in DDLS leads to up regulation of CDK4, increasing cell proliferation. An upregulation of FBN2 in DDLS up regulates TGF-β1, increasing cell proliferation.
Conclusion: Gene expression changes associated with dedifferentiated liposarcomas include changes in cell cycle checkpoint regulation. The signature is associated with poorer overall survival in DDLS and can be used to prognosticate patients.
Citation Format: Deanna Ng, Nicholas Shannon, Johnny Ong, Melissa Teo. 5 Gene prognostic signature for dedifferentiated liposarcomas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2253. doi:10.1158/1538-7445.AM2017-2253
Abstract 2254: Reactive oxygen species modulator 1 (Romo1) predicts unfavorable prognosis in colorectal cancer patients
Authors: Jo Min Jee , Kim Hong Jun , Lee Suk-Young , Lee Dae-Hee , Oh Sang Cheul 
Published: July 2017

Abstract
Reactive oxygen species modulator 1 (Romo1) is a novel protein that has been reported to be crucial for cancer cell proliferation and invasion. However, its clinical implications in colorectal cancer (CRC) patients are not well-known. For the first time, we investigated the association between Romo1 and the clinical outcomes of CRC patients. We examined Romo1 expression in resected tumor tissues immunohistochemically and assessed it with histological scores. We conducted survival analyses for patients who had curative resection (n=190) in accordance with clinical parameters including level of Romo1 expression, and we examined the association between Romo1 expression and cell invasion using Matrigel invasion assay in CRC cell lines. We observed significantly longer mean disease-free survival (DFS) in the low Romo1 group compared with the high Romo1 group (161 vs 127.6 months, p=0.035), and the median overall survival (OS) of the low Romo1 group was significantly longer than that of the high Romo1 group (196.9 vs 171.3 months, p=0.036). Cell invasiveness decreased in the Romo1 knockdown CRC cells in contrast to the controlled cells. Romo1 overexpression in tumor tissue was associated with a high lymph node ratio (LNR) between the metastatic and examined lymph nodes (p=0.025). Romo1 overexpression in tumor tissue was significantly associated with survival in curatively resected CRC patients, suggesting Romo1 expression as a potential adverse prognostic marker. Increased Romo1 expression was found to be associated with high LNR. Cancer invasiveness appeared to be a key reason for the poor survival related to highly expressed Romo1.
Citation Format: Min Jee Jo, Hong Jun Kim, Suk-Young Lee, Dae-Hee Lee, Sang Cheul Oh. Reactive oxygen species modulator 1 (Romo1) predicts unfavorable prognosis in colorectal cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2254. doi:10.1158/1538-7445.AM2017-2254
Abstract 2256: Semaphorin 3C has a possibility of a new prognostic marker in colorectal cancer
Authors: Kawano Shingo , Amemiya Kota , Tsuchiya Yuki , Hagiwara Toshiaki , Matsuzawa Hirokazu , Makino Yurika , Motegi Shunsuke , Tsukamoto Ryoichi , Ichikawa Ryosuke , Kure Kazumasa , Honjo Kumpei , Ro Hisashi , Ito Shingo , Okazawa Yu , Takahashi Rina , Mizukoshi Kosuke , Kawai Masaya , Munakata Shinya , Niwa Koichiro , Ishiyama Shun , Sugimoto Kiichi , Kamiyama Hirohiko , Komiyama Hiromitsu , Takahashi Makoto , Kojima Yutaka , Goto Michitoshi , Okuzawa Atsushi , Tomiki Yuichi , Sakamoto Kazuhiro , Kojima Motohiro , Ochiai Atsushi 
Published: July 2017

Abstract
Background: Generally, Semaphorins are secretary or transmembrane-bound molecules that act as axon guidance cues in the nervous system. Recent research showed increased expression of semaphorin 3C correlates with cancers that possess higher invasive and metastatic characteristics. For example, in breast cancer, inhibition of semaphorin 3C reduces adhesion and invasion. The aim of this study was to evaluate a possibility that semaphorin 3C might be a new prognostic marker in colorectal cancer.
Material and methods: We used two cohorts. Cohort 1 was the 192 patients with colorectal cancer resected surgically between 2009 and 2010. We used GSE 14333 dataset as Cohort 2 which included 226 patients with the colorectal cancer. In each cohorts, we divided the patients in to the two groups i.e., high or low semaphorin 3C expression group using receiver operating characteristic (ROC) curve based on the information of the recurrence of colorectal cancer. Disease-free-survival (DFS) rates were calculated using Kaplan-Meier methods. Differences between curves were evaluated with the log-rank test.
Results: In the cohort 1, the DFS in the patients with higher expression was worse than that in the patients with lower expression (P = 0.0467). Semaphorin 3C has two probes (203788_s_at and 203789_s_at) in GSE14333. In the cohort 2, using each two probe of semaphorin 3C, the DFS in the patients with higher expression was worse than that of those patients with lower expression (P = 0.0015 and P = 0.0003).
Conclusions: This study suggested that Semaphorin 3C might be a new prognostic marker in colorectal cancer.
Citation Format: Shingo Kawano, Kota Amemiya, Yuki Tsuchiya, Toshiaki Hagiwara, Hirokazu Matsuzawa, Yurika Makino, Shunsuke Motegi, Ryoichi Tsukamoto, Ryosuke Ichikawa, Kazumasa Kure, Kumpei Honjo, Hisashi Ro, Shingo Ito, Yu Okazawa, Rina Takahashi, Kosuke Mizukoshi, Masaya Kawai, Shinya Munakata, Koichiro Niwa, Shun Ishiyama, Kiichi Sugimoto, Hirohiko Kamiyama, Hiromitsu Komiyama, Makoto Takahashi, Yutaka Kojima, Michitoshi Goto, Atsushi Okuzawa, Yuichi Tomiki, Kazuhiro Sakamoto, Motohiro Kojima, Atsushi Ochiai. Semaphorin 3C has a possibility of a new prognostic marker in colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2256. doi:10.1158/1538-7445.AM2017-2256
Abstract 2270: C-reactive protein and ovarian cancer risk: preliminary results from the Ovarian Cancer Cohort Consortium (OC3)
Authors: Poole Elizabeth M. , Trabert Britton , Fortner Renée T. , Wentzensen Nicolas , Tworoger Shelley S. 
Published: July 2017

Abstract
Background: C-reactive protein (CRP), a general marker of inflammation has been consistently associated with increased ovarian cancer risk. However, few studies have had the ability to evaluate differences in association by ovarian tumor subtype.
Methods: We pooled existing data on CRP and ovarian cancer risk among nested case-control studies conducted within 6 studies participating in the Ovarian Cancer Cohort Consortium (OC3): CLUEII, EPIC, NHS, NHSII, NYUWHS, and PLCO. Cohort-specific tertiles and quartiles were analyzed using conditional logistic regression, as were common cutpoints at &lt;1mg/L, 1-&lt;10mg/L and ≥10mg/L and Odds Ratios (OR) and 95% confidence intervals (CI) were calculated using random-effects meta-analysis. We also evaluated differences in the association between serous and non-serous cancers.
Results: There was no significant association between the top vs. bottom tertile (OR: 1.10; 95% CI: 0.91-1.32) or the top vs. bottom quartile (OR: 1.10; 95% CI: 0.89-1.36). However, there was a strong, positive association between very high CRP (≥10mg/L vs. &lt;1 mg/L: OR: 1.96; 95% CI: 1.36-2.84). Associations of very high (≥10 mg/L vs. &lt; 1 mg/L) did not differ by histologic type: serous OR: 1.63; 95% CI: 1.11-2.43; endometrioid OR: 1.78; 95% CI: 0.63-5.00; mucinous OR: 10.43; 95% CI: 1.27-85.86; clear cell OR: 2.30; 95% CI: 0.39-13.34).
Conclusion: Our results confirm the consistent observation of a positive association between very high CRP levels and ovarian cancer risk. Pooled analyses accounting for potential confounders are ongoing.
Note: This abstract was not presented at the meeting.
Citation Format: Elizabeth M. Poole, Britton Trabert, Renée T. Fortner, Nicolas Wentzensen, Shelley S. Tworoger. C-reactive protein and ovarian cancer risk: preliminary results from the Ovarian Cancer Cohort Consortium (OC3) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2270. doi:10.1158/1538-7445.AM2017-2270