bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3252: A regulatory variant in CYP2E1 affects the risk of lung squamous cell carcinoma
Authors: Cao Lei , Lin Jia , He Bing , Wang Hongge , Rao Juan , Liu Yingwen , Zhang Xuemei
Published: May 2015

Background: Cytochrome P450 2E1 (CYP2E1), an ethanol-inducible enzyme, has been shown to metabolically activate various carcinogens, which is critical for the development of cancers. It has demonstrated that CYP2E1 polymorphisms alter the transcriptional activity of the gene. However, studies on the association between CYP2E1 PstI polymorphism and non-small cell lung cancer have reported conflicting results. Thus, the gain of the present study was to investigate whether CYP2E1 PstI polymorphism is associated with the development of non-small cell lung cancer in Chinese population.

Method: A case-control study was conducted in which CYP2E1 PstI polymorphism was analyzed in 526 Chinese patients with non-small cell lung cancer and 526 age-matched healthy controls by PCR-RFLP. Odds ratios were estimated by multivariate logistic regression.

Result: For CYP2E1 PstI polymorphism, C allele carriers (OR=0.67; CI=0.51-0.87; P=0.002) was associated with a decreased risk of non-small cell lung cancer when compared with GG homozygotes. In the group analyses by pathological types, for lung SCC and other types the ORs of the C allele carriers were (OR=0.60; CI=0.41-0.88; P=0.009) and (OR=0.54; CI=0.30-0.99; P=0.045), respectively. In the group analyses by smoking status, there was a significant association among nonsmokers (OR=0.57; CI=0.40-0.81; P=0.002), but not smokers. In the group analyses by TNM stage, there was no significance between CYP2E1 PstI polymorphism and the grade of non-small cell lung cancer.

Conclusion: CYP2E1 PstI polymorphism is associated with decreased risk of development of non-small cell lung cancer and lung SCC or other types in Chinese patients.

Key Words: lung cancer; CYP2E1; gene polymorphism; susceptibility; genetic variant

Citation Format: Lei Cao, Jia Lin, Bing He, Hongge Wang, Juan Rao, Yingwen Liu, Xuemei Zhang. A regulatory variant in CYP2E1 affects the risk of lung squamous cell 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 3252. doi:10.1158/1538-7445.AM2014-3252
Abstract 3253: Id4 and FKBP52 interaction regulates androgen receptor activity in normal prostate and prostate cancer
Authors: Joshi Jugal B.
Published: May 2015

The androgen receptor (AR) is a ligand activated transcription factor and important member of the steroid hormone nuclear receptors. Androgen signaling through AR plays a critical role in prostate tumorigenesis. AR transcriptional enhancement normally requires FKBP52 mediated peptidyl-prolyl cis-trans isomerase (PPIase) activity as well as HSP90-binding ability. FK506 binding protein 4 (FKBP52) is an AR folding factor that has critically important physiological roles in male reproductive tissues. Despite several experimental evidences, the molecular mechanism by which FKBP52 promotes AR signaling in prostate cancer still remains to be investigated. Our previous lab studies have demonstrated that Id4 is regulated by androgens in normal prostate epithelial and LNCaP cells. However, the interaction of Id4 and with the androgen receptor pathway in regulating the development and function of the normal prostate still remains elusive. Here we demonstrate that Id4 (inhibitor of differentiation-4) a dominant negative regulator of bHLH transcription factors can physically interact with AR, FKBP52 and HSP90 in early stage prostate cancer LNCaP cells and thereby selectively regulating AR transcriptional activity. Id4 is highly expressed in the normal prostate and decreased in prostate cancer due to promoter hypermethylation. Loss of Id4 in LNCaP cells resulted in ligand independent AR activation characterized by increased AR nuclear translocation and expression of PSA, FKBP51, FKBP52 and ARD1, the transcriptional targets of AR. Differential protein expression between LNCaP and LNCaP-Id4 demonstrated that loss of Id4 cells significantly elevated the levels of FBKP52 and Hsp27, the two known transcriptional regulators and nuclear transporters of AR. More importantly, LNCaP-Id4 cells were able to synthesize significantly higher levels of testosterone as compared to the androgen-dependent LNCaP cells. In conclusion, we demonstrate that loss of Id4 in LNCaP cells elevates FKBP52 levels, transcriptionally potentiates AR activity which in turn then leads to progression of androgen-independent prostate cancer. NIH grant #R01CA128914

Citation Format: Jugal B. Joshi. Id4 and FKBP52 interaction regulates androgen receptor activity in normal prostate and 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 3253. doi:10.1158/1538-7445.AM2014-3253
Abstract 3259: Evaluation of the aromatase inhibition potential of standardized grape extract
Authors: Olson Janet E. , Allen Summer V. , Suman Vera J. , Hoskin Tanya L. , Vachon Celine M. , Ingle James N.
Published: May 2015

OBJECTIVE: To determine the role of grape extract as a potential aromatase inhibitor by testing of plasma hormone levels.

PROCEDURE: A six-week study was conducted involving postmenopausal women during which 94 grams of grape extract was consumed in addition to their usual diet. Plasma hormones were measured before and after treatment.

RESULTS: Of the 18 women involved in the study, average age and BMI were 61.4 years and 24.4, respectively. For the hormone levels studied, the following median (IQR) percent change from baseline to 6 weeks values were found: estradiol +11.8% (-34.4%,+44.2%), p = 0.42; estrone +3.4% (-15.7%,+12.9%), p = 0.64; estrone sulfate +5.3% (-19.9%,+56.3%), p = 0.35; testosterone -1.5% (-14.7%,+10.7%), p = 0.97; androstenedione +12.6% (-17.1%, +49.1%), p = 0.15. The hormone levels did not significantly change between baseline and six weeks. Further, the changes that were observed did not tend to go in the hypothesized direction (estrogens and conjugates increased slightly, testosterone decreased slightly). Only androstenedione showed a trend toward change in the hypothesized direction.

CONCLUSION: In this study, there was no evidence that plasma hormone levels are altered by 6 weeks of daily consumption of 94 grams of grape extract powder.

Citation Format: Janet E. Olson, Summer V. Allen, Vera J. Suman, Tanya L. Hoskin, Celine M. Vachon, James N. Ingle. Evaluation of the aromatase inhibition potential of standardized grape extract. [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 3259. doi:10.1158/1538-7445.AM2014-3259
Abstract 3262: Windows of susceptibility to endocrine disruptors and gene expression in mammary tissue
Authors: Gopalakrishnan Kalpana , Li Qian , Ma Yula , Lambertini Luca , Chen Jia , Teitelbaum Susan L. , Manservisi Fabiana , Belpoggi Fiorella , Bua Luciano , Falcioni Laura
Published: May 2015

Breast cancer is hormonally driven. Endocrine disruptors (EDs) are exogenous substances that mimic or antagonize endogenous hormone action and disrupt endocrine system function. While EDs have been shown to modulate the risk for mammary tumors in animal models, their effects at biologically relevant doses and during critical developmental windows remain poorly understood. We sought to examine the individual and combinatorial effects of three common EDs, diethyl phthalate, methyl paraben and triclosan in rats exposed at five distinct developmental windows of susceptibility - prenatal, neonatal, prepubertal, pubertal and parous/nulliparous. Pathological features and whole genome expression of normal rat mammary tissues were profiled to provide both qualitative and quantitative biological effects of EDs. ED exposure at levels comparable to human exposure resulted in profound changes in both gross phenotypes (e.g. reproductive mortality and mammary gland morphology) as well as in molecular genome profiles. Specifically, ED exposure appeared to hamper normal breast development and resulted in increased mortality in the offspring, possibly due to reduced milk production. Whole genome expression profiling of mammary tissues revealed that in the course of development, the number of differentially expressed genes was lower in ED-exposed rats compared to controls, suggesting developmental delay or suppression by EDs. Pathway enrichment analysis indicated that several processes including inflammation, metabolism and development were disrupted by ED exposure. Taken together, our findings reveal novel mechanisms of ED action in mammary development and strongly suggest the importance of taking into account the timing of exposure when studying the relationship between environment and disease.

Citation Format: Kalpana Gopalakrishnan, Qian Li, Yula Ma, Luca Lambertini, Jia Chen, Susan L. Teitelbaum, Fabiana Manservisi, Fiorella Belpoggi, Luciano Bua, Laura Falcioni. Windows of susceptibility to endocrine disruptors and gene expression in mammary tissue. [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 3262. doi:10.1158/1538-7445.AM2014-3262
Abstract 327: Targeting autophagy overcomes enzalutamide resistance in castrate-resistant prostate cancer cells and improves therapeutic response in a xenograft model
Authors: Yang Joy C. , Nguyen Hao G. , Kung Hsing-Jien , Shi Xu-Bao , Tilki Derya , White Ralph W. deVere , Gao Allen C. , Evans Christopher P.
Published: May 2015

Androgen deprivation or treatment with the androgen receptor signaling inhibitor (ARSI), Enzalutamide (MDV-3100, ENZA) or biclutamide induces autophagy in androgen-dependent and castration resistant CaP (CRPC) cell lines. This autophagic process is mediated by AMPK activation and the suppression of mTOR through Raptor phosphorylation (Serine 792). Using autophagy modulators such as clomipramine (CMI) or metformin in combination with ENZA drastically reduces the cell survival in the colony formation assay. Metformin in combination with ENZA causes AR degradation, which is inhibited by proteasome inhibitor MG132. Transcriptome deep sequencing of parental and ENZA resistant C4-2B cells was carried out to examine differential gene expression pattern that may be related to their ability to survive under constant high exposure to ARSI. In-vivo studies with mice orthotopically implanted with ENZA resistant cells demonstrates that the combination of ENZA and autophagy modulators, CMI or metformin significantly reduces tumor growth when compared to control groups (p<0.005). Blocking autophagy pharmacologically or genetically significantly impairs prostate cancer cell survival in- vitro and in-vivo, implying the therapeutics potential of autophagy inhibitors in the antiandrogen resistance setting.This work is supported in part by Stand Up To Cancer - Prostate Cancer Foundation–Prostate Dream Team Translational Cancer Research Grant. This research grant is made possible by the generous support of the Movember Foundation. Stand Up To Cancer is a program of the Entertainment Industry Foundation administered by the American Association for Cancer Research.

Citation Format: Joy C. Yang, Hao G. Nguyen, Hsing-Jien Kung, Xu-Bao Shi, Derya Tilki, Ralph W. deVere White, Allen C. Gao, Christopher P. Evans. Targeting autophagy overcomes enzalutamide resistance in castrate-resistant prostate cancer cells and improves therapeutic response in a xenograft 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 327. doi:10.1158/1538-7445.AM2014-327
Abstract 3271: The association between germline genetic variants in the PI3K-AKT-mTOR pathway and breast cancer disease free survival
Authors: Pande Mala , Bondy Melissa , Do Kim-Anh , Sahin Aysegul A. , Ying Jun , Mills Gordon B. , Thompson Patricia , Brewster Abenaa
Published: May 2015

Background: PI3K-AKT-mTOR pathway activation in breast tumors is associated with poorer breast cancer disease-free survival and decreased responsiveness to tamoxifen and trastuzumab. However, the role of germline variation in this pathway and breast cancer outcomes has not been previously evaluated. We hypothesized that single nucleotide polymorphisms (SNPs) in candidate genes affecting the PI3K-AKT-mTOR signaling pathway may act as genetic modifiers of breast cancer specific outcomes in patients diagnosed with early stage disease.

Methods: We genotyped 106 SNPs in 13 genes (ADIPOQ, IGF1, INS, IRS1, LEP, LEPR, LEPROT, PIK3CA, PIK3R5, PTEN, TSC1, TSC2, and AKT1) in the P13K-AKT-mTOR pathway in a sample of 1020 women with stages I-II breast cancer treated at the MD Anderson Cancer Center between 1985 and 1999 and for whom long term follow-up on recurrence and death were available. The relationship between SNPs and breast cancer disease free survival (DFS) was assessed using Cox regression analysis. After correcting for false discovery rate (0.10) SNPs significantly associated with DFS in any genetic model (additive, dominant or recessive) were included in multivariable analysis.

Results: After adjusting for race, age at diagnosis, tumor stage and treatment, rs1063539 in ADIPOQ (Hazard ratio [HR] 0.40, 95% confidence interval [CI] 0.19 - 0.86), rs11585329 in LEPROT (HR 0.72, 95% CI 0.53 - 0.98), and rs2519757 in TSC1 (HR 0.29, 95% CI 0.09 - 0.91) were associated with improved DFS and rs1520220 in IGF1 (HR 1.56, 95% CI 1.21 - 2.00) and rs2677760 in PIK3CA (HR 1.43, 95% CI 1.70 - 1.92) were associated with worse DFS. The associations were not modified by type of systemic treatment received. The SNPs were not associated with tumor characteristics such as tumor size, lymph node status, nuclear grade, or hormone receptor status.

Conclusion: In this study, germline allelic variation inthe PI3K-AKT-mTOR pathway appears to modify breast cancer patient outcomes. Future studies are needed to replicate our results and to evaluate the relationship between these polymorphisms and activation of the PI3K-AKT-mTOR pathway in breast tumors.

Citation Format: Mala Pande, Melissa Bondy, Kim-Anh Do, Aysegul A. Sahin, Jun Ying, Gordon B. Mills, Patricia Thompson, Abenaa Brewster. The association between germline genetic variants in the PI3K-AKT-mTOR pathway and breast cancer disease free survival. [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 3271. doi:10.1158/1538-7445.AM2014-3271
Abstract 3274: SNP-SNP interaction analyses of NQO1 and NF-κB signaling pathway genes on breast cancer survival and treatment outcome
Authors: Jamshidi Maral , Fagerholm Rainer , Khan Sofia , Aittomäki Kristiina , Blomqvist Carl , Schmidt Marjanka K. , Nevanlinna Heli ,   BCAC: Breast Cancer Association Consortium
Published: May 2015

To improve individualized therapy, new prognostic and predictive markers are needed to identify patients with expected poor breast cancer outcome and patients who profit most from a given treatment. NF-κB pathway plays a pervasive role in the pathogenesis of cancer through its implication in distinct hallmarks of tumorigenesis. Previous studies have indicated the role of NF-κB transcription factor in promoting cell survival and resistance to chemotherapy in breast carcinoma. We have previously found that NQO1-rs1800566 predicts poor survival among breast cancer patients, particularly after anthracycline-based adjuvant chemotherapy and in patients with p53-aberrant tumors. We have also reported the inverse relationship between NQO1 protein expression and NF-κB activation. We have here analyzed the interaction of SNPs residing within the NF-κB signaling pathway genes, and NQO1-rs6499255 (tagging NQO1-rs1800566 with r2> 0.90) on breast cancer patient survival and treatment outcome. The panel of markers included the NF-κB pathway SNPs available in a custom Illumina iSelect genotyping array designed for Collaborative Oncological Gene-Environment Study (iCOGS) as well as NQO1-rs6499255. For the SNP-SNP interaction analysis on patient survival we conducted a likelihood ratio test that compares Cox's regression model with and without an interaction term to evaluate whether the interaction model is a better fit for the prognostic data. The results obtained from this study will be reported. Identification of interactive SNPs/genes contribute to the elucidation of the mechanisms underlying complex diseases such as breast cancer.

Citation Format: Maral Jamshidi, Rainer Fagerholm, Sofia Khan, Kristiina Aittomäki, Carl Blomqvist, Marjanka K. Schmidt, Heli Nevanlinna, BCAC: Breast Cancer Association Consortium. SNP-SNP interaction analyses of NQO1 and NF-κB signaling pathway genes on breast cancer survival and treatment outcome. [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 3274. doi:10.1158/1538-7445.AM2014-3274
Abstract 3275: Genome-wide association study of childhood body fatness as a risk factor of breast cancer
Authors: Lindstrom Sara , Li Jingmei , Huang Hongyan , Chen Constance , Hunter David J. , Hall Per , Kraft Peter , Tamimi Rulla
Published: May 2015

Girls with large body size have a 15-30% reduced risk of developing breast cancer independent of both birth weight and adult body mass index (BMI). The mechanisms underlying this association are poorly understood, especially since large body size at young age is correlated with earlier pubertal timing, a known risk factor for breast cancer. Self-reported childhood body size, a phenotype which is described in the literature as “childhood body fatness”, has an estimated heritability of 70-80% compared to 40-70% for adult BMI and the correlation between childhood body fatness and adult BMI is only modest (r=0.24-0.30). Still, the literature on genetic associations in pediatric body size-related phenotypes is very sparse.

We conducted a genome-wide association study (GWAS) of childhood body fatness in 10,560 individuals within the Nurses’ Health Study (NHS) and the Health Professional Follow-up Study (HPFS). All subjects were imputed using the 1000 Genomes Phase I v3 reference panel. Analysis was adjusted for top principal components and study/gender. We observed three regions with p<10-7. Two of the regions (TNNI3K and BCDIN3D) have previously been identified as associated with BMI, the former in adolescents. The third region (1p35.2, p=4.8x10-8) constitutes a potential novel locus associated with childhood body fatness. Five additional regions (4 novel) showed suggestive association with childhood body fatness (p<10-6). We are currently replicating these new regions in additional 9,800 subjects from NHS, HPFS and SASBAC, a Swedish breast cancer case-control study. We will also assess the association between identified loci and breast cancer risk. Such analysis will help elucidate the shared genetic component between childhood body fatness and breast cancer and increase our understanding to why early life fatness decreases breast cancer risk.

Citation Format: Sara Lindstrom, Jingmei Li, Hongyan Huang, Constance Chen, David J. Hunter, Per Hall, Peter Kraft, Rulla Tamimi. Genome-wide association study of childhood body fatness as a risk factor of 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 3275. doi:10.1158/1538-7445.AM2014-3275
Abstract 3276: Identifying cross-phenotype inflammatory effects in cancer
Authors: Aquino-Michaels Keston , Trubetskoy Vasya , Im Hae Kyung , Cox Nancy
Published: May 2015

Through genome-wide association studies we now better understand many of the genetic factors contributing to cancer. However, the overlap between cancer associated loci and inflammation associated loci remains poorly understood. Inflammation has long been considered the underlying basis to several complex traits and plays well-established roles in elevating risk to various cancer types. In fact, genome-wide association studies have identified loci associated with inflammatory biomarkers and risk of cancer. Therefore we sought to test cross-phenotype genetic effects between cancer and inflammation biomarkers. While genome-wide significant loci on their own typically account for very little of the measured narrow-sense heritability, polygenic approaches show greater promise to account for the missing heritability of complex traits. Therefore we built polygenic signatures from inflammatory biomarker genome-wide association studies. Preliminary analysis of a case control pancreatic cancer dataset (304 cases/1425 controls) where we applied seven inflammatory signatures indicates that polygenic scores built from IL-10 and C-reactive protein (CRP) are significant predictors of case control status. Importantly the directions of effect for IL-10 and CRP are consistent with the canonical roles of these proteins. We extended this approach to a breast cancer dataset (1082 cases/1086 controls) where we found that polygenic scores built from CRP are significant predictors of case control status. In both the pancreatic and breast cancer datasets CRP polygenic scores decrease risk of case status. This approach will not only inform us of the cancers with overlapping inflammatory effects, but also allow us to identify the components of the immune system that mediate cancer risk. Ultimately this understanding may lead to the use of anti-inflammatory medication for individuals at risk for a particular cancer. Therefore this research addresses cancer prevention by studying the inflammatory genetic component to risk.

Citation Format: Keston Aquino-Michaels, Vasya Trubetskoy, Hae Kyung Im, Nancy Cox. Identifying cross-phenotype inflammatory effects in 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 3276. doi:10.1158/1538-7445.AM2014-3276
Abstract 328: Novel quantitative autophagy analysis by organelle flow cytometry after cell sonication
Authors: Degtyarev Michael , Reichelt Mike , Lin Kui
Published: May 2015

Autophagy is a dynamic process of bulk degradation of cellular proteins and organelles in lysosomes. Current methods of autophagy measurement include microscopy-based counting of autophagic vacuoles (AVs) in cells. We have developed a novel method to quantitatively analyze individual AVs using flow cytometry. This method, OFACS (organelle flow after cell sonication), takes advantage of efficient cell disruption with a brief sonication, generating cell homogenates with fluorescently labeled AVs that retain their integrity as confirmed with light and electron microscopy analysis. These AVs could be detected directly in the sonicated cell homogenates on a flow cytometer as a distinct population of expected organelle size on a cytometry plot. Treatment of cells with inhibitors of autophagic flux, such as chloroquine or lysosomal protease inhibitors, increased the number of particles in this population under autophagy inducing conditions, while inhibition of autophagy induction with 3-methyladenine or knockdown of ATG proteins prevented this accumulation. This assay can be easily performed in a high-throughput format and opens up previously unexplored avenues for autophagy analysis.

Citation Format: Michael Degtyarev, Mike Reichelt, Kui Lin. Novel quantitative autophagy analysis by organelle flow cytometry after cell sonication. [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 328. doi:10.1158/1538-7445.AM2014-328
Abstract 3284: Genome-wide association study identifies new susceptibility loci of epithelial ovarian cancer in Han Chinese
Authors: Li Lian , Chen Kexin , Zang Rongyu , Hao Quen , Hu Zhibin , Lin Dongxin , Ma Ding , Jia Weihua , Shen Hongbing , Wei Qingyi
Published: May 2015

Ovarian cancer is the leading cause of cancer death of gynecological malignancies in the world. To identify risk genetic variants for epithelial ovarian cancer (EOC), we performed a three-stage genome-wide association study (GWAS) in Han Chinese women. We scanned 900,015 single nucleotide polymorphisms (SNPs) in 1,057 EOC cases and 1,191 controls of Chinese women. Forty-one promising SNPs were selected for replication in an independent sample set of 960 EOC cases and 1,799 controls of Chinese women. We further evaluated four significant loci in an additional replication population (492 EOC cases and 1,004 controls of Chinese women) and finally identified two new susceptibility loci at 9q22.33 (rs1413299 in COL15A1, Pmeta = 1.88 × 10-8) and 10p11.21 (rs1192691 near ANKRD30A, Pmeta = 2.62 × 10-8). In addition, two other loci at 12q14.2 (rs11175194 in SRGAP1, Pmeta = 1.14 × 10-7) and 9q34.2 (rs633862 near ABO and SURF6, Pmeta = 8.57 × 10-7) were consistently replicated (P<0.05 in all three stages). These results advance our understanding of the genetic susceptibility to EOC in Han Chinese.

Citation Format: Lian Li, Kexin Chen, Rongyu Zang, Quen Hao, Zhibin Hu, Dongxin Lin, Ding Ma, Weihua Jia, Hongbing Shen, Qingyi Wei. Genome-wide association study identifies new susceptibility loci of epithelial ovarian cancer in Han Chinese. [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 3284. doi:10.1158/1538-7445.AM2014-3284
Abstract 3287: Mutational analysis of serous ovarian cancer using Ion Torrent sequencing
Authors: Masghati Salome , Dorigo Oliver , Santisvkulvong Chintda
Published: May 2015

Ovarian cancer is a biologically diverse tumor with heterogeneous molecular subtypes. Genetic mutations in various oncogenes have been previously identified in high grade serous ovarian cancer, including RB1, CDK12, BRAF, PI3KCA, and KRAS. To further investigate genetic variants, we screened 104 high grade serous ovarian cancer specimens for 2.855 hotspot regions of 50 oncogenes and tumor suppressor genes using the Ion AmpliSeq™ Cancer Hotspot Panel. The panel contains 207 primer pairs in a single tube, using only 10ng of DNA to allow sequencing. Data analysis was performed via Ion Torrent software version 3.0, Nextgene software version 2.3.2, Excel and Medcalc.

Our analysis resulted in the detection of 240 mutation variants. The most frequent mutation variants detected were in PDGFRA (91.35%), FGFR3 (90.38%), APC (86.54%), RET (82.69%), EGFR (74.04%), and TP53 (52.88%). When only COSMIC hotspot mutations were analyzed, genes with frequent mutations were APC (COSM19349; 91.35%),

KDR (COSM149673; 56.7%), PDGFRA (COSM22413; 36.5%), KIT (COSM28026; 19.23%) and TP53 (COSM10704; 7.69%). The COSM19349 APC hot spot mutation results in a frameshift and has been described in colorectal cancer. The hotspot PDGFRA mutation (COSM22413) is a silent mutation that has been previously reported in gastrointestinal stromal tumors, gliomas and endometrial carcinoma. KDR (COSM149673) is a missense mutation that has been found in stomach cancer. KIT (COSM28026) is a missense mutation that has been confirmed as a somatic mutation in chordomas, hematologic malignancies and Merkel cell carcinoma. The TP53 (COSM10704) hotspot mutation has been found in many different tissues, including serous ovarian cancer, breast cancer and GI tract malignancies.

We used the dbNSFP database (database for nonsynonymous SNPs' functional predictions) via Nextgene for functional prediction analysis using six algorithms (PolyPhen, SIFT, Mutation Taster, LRT and 1000 Genomes). Among the most frequent hotspot mutations, TP53 (COSM10704) was found to be ‘damaging’ via all prediction tools.

KIT (COSM28026) was found to be ‘possibly damaging’ via PolyPhen.

In summary, Ion Torrent sequencing revealed mutation variants in serous ovarian cancer that have not been previously identified. Further studies will be needed to analyze the functional and clinical importance of these variants.

Citation Format: Salome Masghati, Oliver Dorigo, Chintda Santisvkulvong. Mutational analysis of serous ovarian cancer using Ion Torrent sequencing. [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 3287. doi:10.1158/1538-7445.AM2014-3287
Abstract 3290: Glucose-regulated protein 58 modulates β-catenin protein stability in cervical adenocarcinoma
Authors: Liao Chia-Jung , Lin Syuan-ling , Wang Tzu-Hao , Lin Kwang-Huei
Published: May 2015

Cervical cancer is still a serious problem threatening to women health worldwide. The incidence of cervical adenocarcinoma (AD) is rising in more developed countries. Previously we have identified that glucose-regulated protein 58 (Grp58) serves as a poor independent prognostic factor for cervical AD. The molecular mechanism underlying Grp58 involved cervical AD carcinogenesis is unknown. The DNA microarray and enrichment analysis were used to identify pathway maps perturbed by knockdown of Grp58 expression. The WNT singling map is one of the significant enriched pathways. The β-catenin, vital effector of WNT signaling, was stably accumulated in Grp58 knockdown stable cells. A membrane fashion of β-catenin was observed in Grp58 knockdown but not control cells. By using transwell assay, we have demonstrated that accumulation of β-catenin, which was induced by Grp58 knockdown or lithium chloride treatment, inhibited the migration ability of HeLa cell. Furthermore, an exclusive expression pattern of Grp58 and β-catenin was observed in cervix tissues. In sum, we have demonstrated that β-catenin stability is negatively regulated by Grp58. Overpression of Grp58 might result in lost or decreased of membranous β-catenin in cervical cancer to promote cancer progression.

Citation Format: Chia-Jung Liao, Syuan-ling Lin, Tzu-Hao Wang, Kwang-Huei Lin. Glucose-regulated protein 58 modulates β-catenin protein stability in cervical adenocarcinoma. [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 3290. doi:10.1158/1538-7445.AM2014-3290
Abstract 3292: Epsin promotes breast cancer progression and metastasis by controlling nf-κb activation
Authors: Cai Xiaofeng , Chen Hong
Published: May 2015

Epsins are a family of endocytic clathrin adaptors. We have previously reported that epsin deficiency inhibits tumor growth by enhancing VEGF signaling in vascular endothelial cells. However, the pattern of tumor-intrinsic expression of epsin and its influence on cancer growth and metastasis is still unclear. Here, we demonstrate that epsin is upregulated in human cancers, including breast cancer, where its high expression correlates with poor relapse-free survival. Epsin promotes cell proliferation and invasion in vitro, and breast tumorigenesis and metastasis in vivo. Mechanistically, epsin interacts with and stabilizes TNFR1 signaling complex in breast cancer cells and enhances NF-κB signaling. Our findings suggest an important role of epsin in contributing to breast cancer progression and metastasis.

Citation Format: Xiaofeng Cai, Hong Chen. Epsin promotes breast cancer progression and metastasis by controlling nf-κb activation. [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 3292. doi:10.1158/1538-7445.AM2014-3292
Abstract 3293: Leptin induces breast cancer metastasis through a Neuropilin-1 (NRP-1)/OBR complex
Authors: Belaid-Choucair Zakia , Dam Julie , Goudin Nicolas , Filhole Odile , Cochet Claude , lepelletier Yves , Coman Tereza , Courtois Geneviève , Colige Alain , Defresne Marie-Paule , Jockers Ralf , Hermine Olivier
Published: May 2015

Several studies have demonstrated Neuropilin-1 (NRP-1) implication in tumor progression independently of its known co-receptors. On the basis of growing connections between breast cancer and obesity, we postulated that leptin and its receptor OBR could be potential candidates. First, using NRP-1 over-expression in NRP-1 negative T47D or NRP-1 silencing in NRP-1 positive MDA-MB231 xenografts, we demonstrated that leptin induced metastasis in lymph node only in NRP-1 positive cells. Second, we demonstrated by co-immunoprecipitation, Proximity Ligation assay (duolink) and BRET analysis that NRP-1 is a co-receptor of OBR. Upon leptin exposure this complex is internalized to the nucleus. At this level, NRP-1 Chip-Seq analysis generated from MDA-MB-231 led to identify sequences of RNA polymerase II (Pol2) and transcription factor binding sites of genes implicated in breast cancer metastasis. This finding was confirmed by the detection of NRP-1/Pol2 interaction and by transcriptome analysis of RNA extracts from T47D-NRP-1 xenografts that shows modulation of these genes.

In conclusion, we have demonstrated that NRP-1 is a co-receptor of OBR and upon leptin exposure may play a role as a transcription factor regulating genes involved in cancer progression. Thus, NRP-1 might be a therapeutic target to prevent cancer metastasis induced by leptin

Citation Format: Zakia Belaid-Choucair, Julie Dam, Nicolas Goudin, Odile Filhole, Claude Cochet, Yves lepelletier, Tereza Coman, Geneviève Courtois, Alain Colige, Marie-Paule Defresne, Ralf Jockers, Olivier Hermine. Leptin induces breast cancer metastasis through a Neuropilin-1 (NRP-1)/OBR complex. [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 3293. doi:10.1158/1538-7445.AM2014-3293
Abstract 3294: Notch signaling is regulated by vacuolar ATPase in triple negative breast cancer
Authors: Pamarthy Sahithi , Beaman Kenneth D.
Published: May 2015

Triple Negative Breast Cancer (TNBC) constitutes 15 % of all diagnosed breast cancers. It is more common among African American and younger women. TNBC cells do not express Estrogen, Progesterone and Her2 receptors. As a result, patients have limited treatment options. To effectively treat TNBC, it is important to identify and target specific signaling pathways that contribute to TNBC tumorigenesis.

Notch signaling is an evolutionarily conserved juxtacrine pathway that is important in normal mammary development and has been shown to play a role in TNBC progression. Although there has been considerable research on role of Notch in TNBC, little is known about its regulation. Identifying novel molecules that can regulate Notch can lead to the development of effective combinatorial treatments.

The Vacuolar ATPase (V-ATPase) has been shown to regulate Notch Signaling in Drosophila. V-ATPase is an ATP driven proton pump, which is responsible for maintaining pH. Tumor cells increase expression of V-ATPase, which aids in cancer proliferation and metastasis.

Here, we report that the a2 isoform of V-ATPase regulates Notch Signaling in breast cancer. Among isoforms a1, a2, a3 and a4, breast cancer cell lines robustly expressed a2 V-ATPase on their surface. a2 V-ATPase is present towards the leading edge of cells and is higher in TNBC cell lines (MDA-MB-231, MDA-MB-468) as compared to normal chemically transformed cell line (184A1) and hormonal positive cell line (MCF7). Expression of Notch ligands, receptors and target genes was also higher in TNBC cell lines as compared to Non-TNBC cell lines. siRNA mediated knockdown of a2 V-ATPase resulted in an increase of Notch Signaling in the Triple Negative MDA-MB-231 cell line. To understand the molecular mechanism behind this regulation, we used Bafilomycin to inhibit V-ATPase activity. Following V-ATPase inhibition, the expression of Jagged1, a Notch ligand was increased. Our results suggest that Vacuolar ATPase can regulate Notch Signaling by affecting ligand internalization through endocytosis. We further assessed the effect of V-ATPase and Notch signaling inhibitors on cell proliferation of the breast cancer cell lines (184A1, MCF7, MDA-MB-231, MDA-MB-468). Combined inhibition of V-ATPase and Notch signaling caused significant cell death in all cell lines tested. Together our results identify a2 V-ATPase as a novel regulator of Notch signaling in Breast Cancer and that targeting both V-ATPase and Notch Signaling can be beneficial for Triple Negative Breast Cancer.

Citation Format: Sahithi Pamarthy, Kenneth D. Beaman. Notch signaling is regulated by vacuolar ATPase in triple negative 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 3294. doi:10.1158/1538-7445.AM2014-3294
Abstract 3295: S6 kinase signaling in prognosis and tamoxifen response in two randomized breast cancer cohorts
Authors: Bostner Josefine , Karlsson Elin , Bivik Cecilia , Perez-Tenorio Gizeh , Fornander Tommy , Stål Olle
Published: May 2015

Detecting signals in the mammalian target of rapamycin (mTOR), and the estrogen receptor (ER) pathways for prediction of treatment response may be a future clinical tool in primary breast cancer. Here, we investigated the validity and value of the mTOR targets p70-S6 kinase (S6K) 1 and 2 as biomarkers for tamoxifen sensitivity in vitro and in two independent tamoxifen randomized postmenopausal breast cancer cohorts. In addition, the prognostic value of the S6Ks was evaluated.

We found S6K1 to correlate with HER2 and cytoplasmic Akt activity, whereas S6K2 and phosphorylated S6K were closer connected with ER positivity, low proliferation and nucleic p-Akt. Treatment prediction and prognosis were evaluated by immunohistochemical staining. Nuclear accumulation of S6K1 was indicative of a reduced tamoxifen treatment effect, compared with a significant benefit from tamoxifen treatment in patients without tumor S6K1 nuclear accumulation. Patients with a combination of S6K1 nuclear accumulation and S6K2 cytoplasmic accumulation in the tumor cells had no tamoxifen benefit. Also, S6K1 and S6K2 activation, indicated by p-S6K-t389 expression, was associated with low benefit from tamoxifen compared with untreated patients. In addition, high protein expression of S6K1, independent of localization, predicted worse prognosis. This was not evident for variations in S6K2 or p-S6K-t389 expression.

In conclusion, the mTOR targeted kinases S6K1 and S6K2 interfere with proliferation and response to tamoxifen. Monitoring their activity and intracellular localization may provide biomarkers for breast cancer treatment, allowing for identification of a group of patients less likely to benefit from tamoxifen and thus in need of an alternative or additional treatment.

Citation Format: Josefine Bostner, Elin Karlsson, Cecilia Bivik, Gizeh Perez-Tenorio, Tommy Fornander, Olle Stål. S6 kinase signaling in prognosis and tamoxifen response in two randomized breast cancer cohorts. [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 3295. doi:10.1158/1538-7445.AM2014-3295
Abstract 3300: Estrogen stimulates cell cycle progression dependent on Src kinase activity and IMP1 stabilization of myc mRNA in estrogen receptor-positive breast cancer cells
Authors: Abdullah Christopher , Korkaya Hasan , Courtneidge Sara A.
Published: May 2015

Estrogen (E2) stimulation promotes proliferation in estrogen receptor-positive (ER+) breast cancer, which accounts for 60-75% of all breast cancer. E2 signals through the ER via three pathways: the classical genomic pathway involving transcriptional activation of the ER, crosstalk between ER and other transcription factors, and “non-genomic” signaling cascades, which do not involve the transcriptional activity of the ER. This “non-genomic” pathway likely involves Src family kinases (SFKs) and has been implicated in cell cycle progression in fibroblasts engineered to express the ER (Castoria 1999). While not typically amplified or mutationally activated in ER+ breast cancer, Src is hyperactive in most breast cancers.

To test whether SFK activity is required for G1/S cell cycle progression in breast cancer cells, we pretreated quiescent ER+ MCF7 cells with a selective SFK inhibitor (SU11333) prior to stimulation with E2. Inhibition of SFK activity blocked progression from G1 to S phase. Previous data from our laboratory has demonstrated that G1/S progression in response to peptide growth factor stimulation of quiescent fibroblasts is dependent upon SFK-mediated stabilization of myc mRNA (Bromann 2005). We therefore tested whether SFK activity also regulates myc expression in breast cancer cells. Our data suggest that SFK activity is required for accumulation of myc mRNA in quiescent MCF7s after E2 stimulation, and that E2 stimulated stabilization of myc mRNA is dependent on SFK activity.

One potential myc mRNA stabilizer is IMP1 (IGF2BP1, ZBP1, CRDBP) an RNA binding protein that is primarily expressed during development and in stem cells, and re-expressed in some cancer types, including breast cancer. IMP1 has previously been shown to stabilize myc mRNA in stem cells as well as osteosarcoma cells. While the signaling pathways that mediate this stabilization have not been well studied, previous work has identified a Src phosphorylation site on IMP1 which regulates IMP1 function (Sasaki 2010). We hypothesized that IMP1 may play a role in E2-mediated stabilization of myc mRNA. Indeed, MCF7 cells require IMP1 to accumulate myc mRNA after E2 stimulation. In summary, E2 stimulation activates SFKs, which then promote G1/S cell cycle progression in part by stabilizing myc mRNA levels, through the activity of IMP1.

Bromann et al JBC, 2005

Castoria et al. EMBO J, 1999

Sasaki et al. J Neurosci, 2010

Citation Format: Christopher Abdullah, Hasan Korkaya, Sara A. Courtneidge. Estrogen stimulates cell cycle progression dependent on Src kinase activity and IMP1 stabilization of myc mRNA in estrogen receptor-positive 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 3300. doi:10.1158/1538-7445.AM2014-3300
Abstract 3301: Inhibition of the secretory pathway calcium ATPase 1 (SPCA1) in MDA-MB-231 breast cancer cells and the effect on protein expression
Authors: Lee Jane M. , Ross Diana G. , Monteith Gregory R. , Roberts-Thomson Sarah J.
Published: May 2015

SPCA1 is a Ca2+ pump located on the membrane of the Golgi apparatus and is involved in controlling calcium levels within the Golgi. We have previously shown that SPCA1 is upregulated in some basal-like breast cancers. Given the role of the Golgi in protein trafficking and processing, we hypothesised that SPCA1 may be involved in regulating the expression of proteins important in cancer. To address this hypothesis we used 2D-DIGE to identify proteins that are altered when SPCA1 is silenced in MDA-MB-231 basal cancer cells. MDA-MB-231 cells were seeded into 6-well plates (75,000 cells/well) and treated with SPCA1 or non-targeting siRNA. Protein was isolated 72 h post siRNA treatment, and SPCA1 silencing was confirmed by immunoblotting. 2D-DIGE and MS/MS were used to identify proteins with differential expression with SPCA1 silencing. Heat shock protein 60 (HSP60) was identified as a protein potentially sensitive to SPCA1 silencing using 2D-DIGE. Immunoblotting and real time RT-PCR showed that SPCA1 silencing reduced HSP60 protein and mRNA expression by 81±2% and 60±7% (n=3, P<0.05), respectively. Given the role of HSP60 in cell death pathways, we also assessed the consequences of SPCA1 silencing on heat shock and staurosporine-induced cell death. SPCA1 silencing had no effect on heat shock-induced cell death but reduced the sensitivity of MDA-MB-231 breast cancer cells to staurosporine. Although further studies are required to fully characterize the functional consequences of HSP60 downregulation upon SPCA1 silencing, this work suggests that SPCA1 is a complex regulator of proteins important in cancer pathways.

Citation Format: Jane M. Lee, Diana G. Ross, Gregory R. Monteith, Sarah J. Roberts-Thomson. Inhibition of the secretory pathway calcium ATPase 1 (SPCA1) in MDA-MB-231 breast cancer cells and the effect on protein expression. [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 3301. doi:10.1158/1538-7445.AM2014-3301
Abstract 3307: Angiomotin regulates the Hippo pathway to increase prostate cancer cell growth
Authors: Ortiz Angelica , Zeng Hao , Lin Sue-Hwa
Published: May 2015

Angiomotin (AMOT) is a member of the motin family that consists of three members, AMOT, AMOTL1, and AMOTL2, which function as either positive or negative regulators of cell growth depending on interacting proteins and cellular context. The role of AMOT in prostate cancer (PCa) has not been studied. AMOT is expressed as two isoforms, AMOTp80 and AMOTp130. The AMOTp130 isoform has an extended 409 aa N-terminal domain that is absent in AMOTp80. Both AMOTp80 and AMOTp130 are expressed in LNCaP and C4-2B4, but at a very low level in PC3-mm2 PCa cells. We found that expression of AMOTp130 and AMOTp80 have distinct functions in PCa cells. While AMOTp80 functions as a tumor promoter by enhancing PCa cell proliferation, AMOTp130 did not. We further show that AMOTp80 affects the Hippo tumor suppressor pathway by inhibiting LATS1 and YAP phosphorylation, resulting in nuclear translocation of YAP. Once in the nucleus, YAP increases transcription of the target protein BMP4. Moreover, inhibition of BMP receptor activity by LDN-193189 abrogates AMOTp80-mediated proliferation. These observations suggest that AMOTp80 induces the expression of BMP4, which functions in an autocrine manner to promote PCa cell proliferation. Together, our studies identify AMOTp80-MST-LATS1-YAP-BMP4 as a novel signal pathway that regulates PCa cell proliferation.

Citation Format: Angelica Ortiz, Hao Zeng, Sue-Hwa Lin. Angiomotin regulates the Hippo pathway to increase prostate cancer cell growth. [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 3307. doi:10.1158/1538-7445.AM2014-3307