bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1736: Mitochondrial priming of apoptotic machinery in pancreatic cancer by BPM 31510 to enhance efficacy of chemotherapy.
Authors: Narain Niven R. , Walshe Tony E. , Lee Arleide , Ouro-Djoba Rakib , Mauro Lucia , Yunis Adel , Vishnudas Vivek K. , Sarangarjan Rangaprasad , Jimenez Joaquin J.
Published: August 2014

Recent evidence suggests that the cytotoxic effects of chemotherapy, specifically in cancer cells is a consequence of “pre-treatment priming” event wherein exposure to the chemotherapeutic agents pushes the cancer cells beyond an apoptotic threshold to effectuate cell death. (Letai, A et. al, Science. 2011 Nov 25;334 (6059) BPM 31510 is a ubidecarenone containing proprietary formulation that effectuates a metabolic switch from glycolysis towards enhanced mitochondrial oxidative phosphorylation resulting in the recapitulation of apoptosis in cancer. The current study was aimed at investigating whether pretreatment with BPM 31510 results in mitochondrial priming, thereby augmenting the cytotoxic effect of standard of care chemotherapeutic agents. The human derived pancreatic cancer cell line Mia-Paca-2 was subjected to either (a) pretreatment with BPM 31510 followed by co-treatment with gemcitabine or (b) co-treatment with BPM 31510 and gemcitabine. BPM 31510 exposure resulted in decreased cell proliferation rates that were significantly higher than gemcitabine alone in Mia-Paca-2 cell line. Pre-treatment with BPM 31510 augmented the cytotoxic potential of gemcitabine, a phenomenon that was also observed in the co-treatment regimen. In vivo pancreatic cancer model using Mia-Paca-2 cell lines were developed to further investigate the priming effect of BPM 31510. Results demonstrate that 1-week and 2-week pretreatment with intravenous BPM 31510 followed by gemcitabine results in improved survival in the pancreatic cancer model compared to the co-treatment regimen. The data demonstrates that addition of BPM

31510 synergizes the cytotoxic effect of gemcitabine in pancreatic cancer. In addition, pretreatment with BPM 31510 followed by gemcitabine treatment is associated with improved survival. Taken together, the data suggest that BPM 31510 may be a viable mitochondrial priming agent to sensitize cancer cells to the cytotoxic effects of gemcitabine in pancreatic cancer.

Citation Format: Niven R. Narain, Tony E. Walshe, Arleide Lee, Rakib Ouro-Djoba, Lucia Mauro, Adel Yunis, Vivek K. Vishnudas, Rangaprasad Sarangarjan, Joaquin J. Jimenez. Mitochondrial priming of apoptotic machinery in pancreatic cancer by BPM 31510 to enhance efficacy of chemotherapy. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1736. doi:10.1158/1538-7445.AM2013-1736
Abstract 1737: Pharmacological inhibition of fatty acid synthase regulates BH3-only proteins and sensitizes breast cancer cells to a small molecule Bcl-2/Bcl-xL inhibitor to induce apoptosis.
Authors: Venkatapoorna Chandra Mohan Kurapaty , Espinoza Ingrid , Kaufmann Scott H , Lupu Ruth
Published: August 2014

Fatty acid synthase (FASN), the key enzyme in de novo lipogenesis, is significantly upregulated and activated in many carcinomas, including breast cancer. Pharmacological inhibition of FASN induces apoptosis of FASN expressing cancer cells. However, the mechanism is still unclear. Inhibition of FASN in breast cancer cells induces a dose dependent increase in apoptotic cell death with a concomitant upregulation of the pro-apoptotic BH3-only proteins Noxa, Bim and Puma (mRNA and protein). Blockage of the BH3-only protein expression using specific small interfering ribonucleic acid (siRNA), significantly diminishes the apoptotic effect induced by the inhibition of FASN. Moreover, addition of palmitate, the end product of FASN, protects the cells from the proapoptotic effects of FASN inhibition by suppressing expression of Noxa, Bim and Puma. These data indicate that Noxa, Bim and Puma are essential for the apoptotic effect induced by the inhibition of FASN. Combination therapeutic regimens have received growing attention for the possibility of higher efficacy with reduced toxicity. Thus, we investigated the combinatorial effect of pharmacological inhibition of FASN and Bcl-2/Bcl-xL. While a small molecule Bcl-2/Bcl-xL inhibitor alone did not induce apoptosis, co-treatment of the small molecule Bcl-2/Bcl-xL inhibitor with a synthetic FASN inhibitor significantly surpassed the apoptotic effect of the synthetic FASN inhibitor alone. Accordingly, combined treatment of a FASN inhibitors and Bcl-2/Bcl-xL antagonists represents a new rational combination that merits further consideration in FASN overexpressing tumors.

Citation Format: Chandra Mohan Kurapaty Venkatapoorna, Ingrid Espinoza, Scott H Kaufmann, Ruth Lupu. Pharmacological inhibition of fatty acid synthase regulates BH3-only proteins and sensitizes breast cancer cells to a small molecule Bcl-2/Bcl-xL inhibitor to induce apoptosis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1737. doi:10.1158/1538-7445.AM2013-1737

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 1738: Multiparametric approaches to evaluate mitochondrial toxicity, oxidative stress and apoptosis in drug discovery studies.
Authors: Gillis Katherine , Clor Julie , Tran Kimvan , Khan Asima , Tyagarajan Kamala
Published: August 2014

Drug discovery and toxicity studies require access to methods and technologies that can provide comprehensive evaluation of the types of cellular impacts caused by anti-cancer compounds. In particular, there is growing emphasis on the study of mitochondrial toxicity at an early phase of compound evaluation and its relations to multiple cell death processes that include apoptotic cell death, caspase-independent death and autophagic processes. The study of oxidative stress and its relation to cell death pathways also provides insights into mechanism of cellular stress. In this study we have used multiparametric assays that correlate changes in mitochondria with different cell health parameters using plate-based microcapillary cytometry on guava easyCyte platforms. Compound screening studies were performed on Jurkats (suspension) and HeLa adherent cell lines using an array of over 80 compounds to study inter-relationships between mitochondrial perturbations and different cell health parameters. Compounds were evaluated for mitochondrial depolarization and relationship to apoptosis and cell death; mitochondrial perturbations with regard to caspase levels were also evaluated; and the parallel connection between mitochondrial superoxide stress and apoptosis was also studied. Our studies identified several hit compounds, examples of which included compounds like gambogic acid, thimerosal, camtothecin, antimycin A, staurosporine, celastrol and juglone which caused significant mitochondrial perturbation and apoptosis. In addition, compounds such as antimycin A and staurosporine caused significant oxidative stress. Simplified and rapid screening assays that provide information on multiple parameters of cellular stress and apoptosis can greatly benefit the identification of hit compounds and provide a more comprehensive understanding of the mechanism of compound toxicity.

Citation Format: Katherine Gillis, Julie Clor, Kimvan Tran, Asima Khan, Kamala Tyagarajan. Multiparametric approaches to evaluate mitochondrial toxicity, oxidative stress and apoptosis in drug discovery studies. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1738. doi:10.1158/1538-7445.AM2013-1738
Abstract 1740: SATB2 localizes to mitotic microtubules and centrosome of cell cycles and regulates G1 phase cell cycle via proteosomal-dependent CDK2 in SKOV-3 ovarian cancer cells.
Authors: Sin Eun Ah , Sohn Eun J. , Jung Ji Hoon , Lee Duckgue , Kim Bonglee , Jung Duck-Beom , Kim Ji-Hyun , Lee Hyo-Jeong , Kim Sung Hoon
Published: August 2014

Though special AT-rich sequence-binding protein 2 (SATB2) is known as a transcriptional regulator of skeletal development and osteogenic differentiation, the underlying role of SATB2 still remains unclear. In the present study, we found that SATB2 localized to the mitotic microtubules, centrosome and midbodies in mitotic cells with α-tubulin. SATB2 knockdown by siRNA transfection reduced the cell viability, and increased the number of ethidium homodimer positive stained cells in SKOV3 ovarian cancer cells. Consistently, cell cycle analysis revealed that SATB2 knockdown accumulated G1 arrest and downregulated the cyclin-dependent kinase 2 (CDK2) expression by western blotting, while immunofluorescence assay revealed that CDK2 and SATB2 were co-localized in SKOV3 cells. Furthermore, proteosomal inhibitor MG132 treatment reversed the downregulation of CDK2 induced by SATB2 knockdown in SKOV3 cells. Overall, our findings suggest that SATB2 regulates the mitosis and G1 phase cell cycle via proteosomal ubiquitination in SKOV3 ovarian cancer cells.

Citation Format: Eun Ah Sin, Eun J. Sohn, Ji Hoon Jung, Duckgue Lee, Bonglee Kim, Duck-Beom Jung, Ji-Hyun Kim, Hyo-Jeong Lee, Sung Hoon Kim. SATB2 localizes to mitotic microtubules and centrosome of cell cycles and regulates G1 phase cell cycle via proteosomal-dependent CDK2 in SKOV-3 ovarian cancer cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1740. doi:10.1158/1538-7445.AM2013-1740
Abstract 1754: Dormant cancer cells are resistant to conventional therapies and induce tumor vessels after chemotherapy.
Authors: Yano Shuya , Mii Sumiyuki , Tome Yasunori , Hiroshima Yukihiko , Uehara Fuminari , Miwa Shinji , Fujiwara Toshiyoshi , Hoffman Robert M.
Published: August 2014

Dormant cancer cells were visualized with a fluorescence ubiquitination cell cycle indicator (FUCCI). FUCCI-expressing xenografts were established from MKN45 gastric cancer cells. Within 7 days after inoculation, FUCCI-expressing nascent tumors contained a mixture of cancer cells in G0/G1 phase and in S/G2/M phases. Nascent tumors treated with cyto-toxic agents showed that cancer cells in G0/G1 phase can survive and become dormant. As tumors grew, the proliferating area was only at the surface and the dormant area predominated. Chemotherapy killed only the minority population of proliferating cancer cells at the surface and had little effect on the majority of dormant cancer cells in the tumor center. Furthermore, we investigated tumor chemoresistance using nestin-driven GFP transgenic mice that have GFP-expressing nascent tumor vessels. Chemotherapy-treated tumors had much more and deeper tumor vessels than control tumors, suggesting that dormant cancer cells may protect themselves by inducing tumor vessels. This report suggests that dormant cancer cells play a large role in drug resistance.

Citation Format: Shuya Yano, Sumiyuki Mii, Yasunori Tome, Yukihiko Hiroshima, Fuminari Uehara, Shinji Miwa, Toshiyoshi Fujiwara, Robert M. Hoffman. Dormant cancer cells are resistant to conventional therapies and induce tumor vessels after chemotherapy. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1754. doi:10.1158/1538-7445.AM2013-1754
Abstract 1757: Gelfoam® histoculture enables cancer cells to mimic in vivo cell cycle behavior.
Authors: Yano Shuya , Mii Sumiyuki , Tome Yasunori , Hiroshima Yukihiko , Uehara Fuminari , Miwa Shinji , Hoffman Robert M.
Published: August 2014

Tumors in vivo are well known to behave quite differently from cancer cells cultured on plastic or glass plates (monolayer culture). Therefore, cancer cell-cycle kinetics were visualized in Gelfoam® 3D-histoculture using HeLa cells and MKN45 cells expressing fluorescence ubiquitination cell cycle indicator (FUCCI), whereby quiescent cancer cells express mKusabira-Orange fluorescent protein (red) and proliferating cells express mAzami-Green fluorescent protein (green). Gelfoam® histoculture of FUCCI-expressing cancer cells results in tumor-like structures with quiescent cancer cells in the center and proliferating cells at the surface, similar to what occurs in vivo as visualized by FUCCI confocal microscopy. These results have profound implications for cell cycle studies, since most cellular and molecular studies of the cancer cell cycle are performed on plastic monolayer culture where characteristics of the cells may be quite different from the tumor in vivo.

Citation Format: Shuya Yano, Sumiyuki Mii, Yasunori Tome, Yukihiko Hiroshima, Fuminari Uehara, Shinji Miwa, Robert M. Hoffman. Gelfoam® histoculture enables cancer cells to mimic in vivo cell cycle behavior. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1757. doi:10.1158/1538-7445.AM2013-1757
Abstract 1764: Overexpression of the DNA repair regulator PARI in pancreatic cancers promotes genomic instability and tumorigenesis.
Authors: O'Connor Kevin , Dejsuphong Donniphat , D'Andrea Alan , Moldovan George-Lucian
Published: August 2014

Inappropriate homologous recombination (HR) causes genomic instability and cancer. We recently showed that the protein PARI, containing a UvrD-like helicase domain, is a novel PCNA interacting partner, required for preservation of genome stability in human and DT40 chicken cells (Moldovan et al, Molecular Cell 2012). Using cell-based and biochemical assays, we showed that PARI restricts unscheduled recombination by interfering with the formation of RAD51-DNA HR structures. Thus, we proposed that PARI is a long sought-after factor that suppresses inappropriate recombination events at mammalian replication forks.

We recently found that PARI is specifically overexpressed in pancreatic cancer cells (O'Connor et al, Cancer Research, in revision). PARI up-regulation results in DNA repair deficiency and genomic instability. Importantly, PARI knockdown compromises the proliferation of pancreatic cancer cells in vitro. PARI depletion results in an altered cell cycle, defined by S-phase accumulation, perturbed DNA replication, and activation of the DNA damage response pathway in the absence of exogenous DNA damage. Moreover, we found that in pancreatic cancer cells, PARI overexpression results in DNA damage tolerance by promoting replication of damaged DNA. Finally, using a xenograft tumor model in nude mice we show that PARI knockdown reduces pancreatic tumor growth in vivo. Thus, our work suggests that PARI is a potential target for pancreatic ductal adenocarcinoma therapy.

Citation Format: Kevin O'Connor, Donniphat Dejsuphong, Alan D'Andrea, George-Lucian Moldovan. Overexpression of the DNA repair regulator PARI in pancreatic cancers promotes genomic instability and tumorigenesis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1764. doi:10.1158/1538-7445.AM2013-1764
Abstract 1785: Ubiquitin specific protease 3 is a regulator of histone ubiquitination required for maintenance of genomic stability.
Authors: Sharma Nidhi , Zhu Qianzheng , Wang Qi-En , Wani Altaf A.
Published: August 2014

The ubiquitination of histones is crucial for the maintenance of genomic stability. Little is understood about the deubiquitinating enzymes that catalyze the removal of ubiquitin from histones in response to UV damage. In this study, we identify the ubiquitin-specific protease 3 (USP3) as a deubiquitinating enzyme which catalyzes the cleavage of ubiquitin from γH2AX and uH2A in response to UV damage. Using GFP-tagged USP3 we demonstrate that USP3 is a nuclear protein. To determine the function of USP3 in chromatin regulation we ectopically expressed USP3 in Hela cells and analyzed the ubiquitination status of histones. Our analysis demonstrated that the overexpression of the wild type USP3 (USP3wt), but not the catalytically inactive mutant (USP3C1685) resulted in a decrease in the ubiquitination of γH2AX and uH2A. This result was further supported by the functional ablation of USP3 by RNAi. The efficient reduction of the USP3 protein was accompanied by a significant increase in the levels of uH2A and the ubiquitination of γH2AX. In addition, we were also able to demonstrate that the depletion of USP3 increases the persistence of uH2A and Fk2 foci formation in response to UV damage. Together, our results implicate that USP3 is involved in the removal of ubiquitin marks from chromatin-bound proteins as a means to attenuate the damage signal. Thus, in the presence of UV damage, the functional ablation of USP3 may lead to a delay in cell cycle progression and the activation of checkpoint signals, which in turn could induce genotoxic stress. This work was supported by the PHS grant ES2388, ES12991 and CA93413.

Citation Format: Nidhi Sharma, Qianzheng Zhu, Qi-En Wang, Altaf A. Wani. Ubiquitin specific protease 3 is a regulator of histone ubiquitination required for maintenance of genomic stability. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1785. doi:10.1158/1538-7445.AM2013-1785
Abstract 1786: UV damage-induced histone H3 lysine 56 deacetylation regulates nucleotide excision repair and checkpoint signaling.
Authors: Battu Aruna , Wani Altaf Ahmad
Published: August 2014

Nucleotide excision repair (NER) is the major pathway in cells for the removal of bulky photoproducts and ensuring integrity of the genome. NER constitutes lesion recognition, excision of damaged DNA, synthesis of a DNA fragment, and ligation of the repair patch. Nonetheless, DNA within the cells is compacted into chromatin and lesion recognition requires chromatin to be remodeled to permit access of repair and checkpoint machinery to the damage site. Consequent to repair of the lesions, chromatin structure is restored to its original state so that cells resume cell cycle progression. Although several reports have demonstrated the role of chromatin remodelers, histone modifications and histone chaperones in DNA damage response (DDR), knowledge of how these epigenetic mechanisms control the DDR is largely in its infancy. We and others have recently observed that histone H3 is deacetylated on lysine 56 immediately in response to UV-induced DNA damage. Acetylation is promptly restored upon completion of repair and is necessary for recovery from DNA damage induced cell cycle arrest. However, the function of H3K56 deacetylation in the DDR remains to be determined. We found that UV-induced deacetylation of H3K56 is abrogated in the presence of TSA and sodium butyrate, both of which inhibit class I histone deacetylases (HDACs). Knockdown of either HDAC1 or HDAC2 alone did not affect UV-induced deacetylation of H3K56. However, simultaneous knockdown of both HDAC1 and 2 resulted in retention of H3K56 acetylation after UV irradiation. Consistently, both HDAC1 and HDAC2 localized to sites of UV damage. These observations are consistent with that reported by Miller et. al., indicating that HDAC1 and HDAC2 are responsible for the deacetylation of H3K56 in response to DNA damage. Since the deacetylation of H3K56 is one of the most immediate responses to UV-irradiation, we assumed it to play a crucial role in NER. Accordingly, we observed that CPD removal was affected in H1299 cells with histone H3K56Q mutation that mimics constitutive acetylation. Furthermore, we found that localization of the checkpoint kinase ATM to UV damage sites was affected in these cells. Consequently, ATM-mediated H2AX phosphorylation was also found to be defective in the mutant cells. Based on our observations, we speculate that HDAC1 and 2 mediated H3K56 deacetylation regulates NER and checkpoint signaling cascade in response to UV damage.

This work was supported by PHS grants ES2388, ES12991 and CA93413 and Pelotonia postdoctoral fellowship to AB.

Citation Format: Aruna Battu, Altaf Ahmad Wani. UV damage-induced histone H3 lysine 56 deacetylation regulates nucleotide excision repair and checkpoint signaling. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1786. doi:10.1158/1538-7445.AM2013-1786
Abstract 1797: Differential gene expression profiles of FFPE samples with SensationPlus™ and GeneChip™ arrays.
Authors: Chamnongpol Sangpen , Ujvari Andrea , Kourennaia Olga , Myers Kim , Bowers Lisa , Fisher Jesse , Post Marc , Kubu Chris
Published: August 2014

Formalin-fixed and paraffin-embedded (FFPE) tissue samples represent a largely untapped wealth of information on the transcriptional states of clinically archived materials for retrospective studies. Although the RNA is degraded during fixation and storage and has been considered unusable, Affymetrix has developed SensationPlus™, a target-preparation method that allows transcriptional profiling from 20 ng of FFPE total RNA using GeneChip™ arrays. We will demonstrate that SensationPlus™ expression profiles from decade old FFPE materials are extremely similar to data from their matched fresh frozen (FF) RNA counterparts.

Citation Format: Sangpen Chamnongpol, Andrea Ujvari, Olga Kourennaia, Kim Myers, Lisa Bowers, Jesse Fisher, Marc Post, Chris Kubu. Differential gene expression profiles of FFPE samples with SensationPlus™ and GeneChip™ arrays. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1797. doi:10.1158/1538-7445.AM2013-1797
Abstract 1805: LSD1 regulation of SKP2 expression in prostate cancer.
Authors: Ding Liya , Huang Haojie
Published: August 2014

Prostate carcinogenesis is a complicated process involving both genetic and epigenetic changes. Aberrant expression of LSD1 (Lysine-specific demethylase 1) has been reported in human prostate cancers. Although high expression of LSD1 is implicated in prostate tumorigenesis, the underlying molecular mechanisms are not fully understood. Here we reported that LSD1 promotes proliferation and migration of prostate cancer cells via up-regulation of SKP2 expression. Knockdown of LSD1 using small interfering RNA (siRNA) decreases SKP2 expression and inhibits prostate cancer cell proliferation and migration. Promoter luciferase reporter and chromatin immunoprecipitation (ChIP) experiments show that LSD1 regulates SKP2 expression by controlling the levels of H3K4 methylation in SKP2 promoter and this function of LSD1 is mediated through its demethylase activity. As demonstrated by immunohistochemistry, LSD1 and SKP2 proteins positively correlate with each other and their expression is associated with cell proliferation in human prostate cancer tissues. Our data not only identify SKP2 as a direct regulated target gene of LSD1 but also reveal that SKP2 plays an important role in contributing LSD1 mediated prostate cancer cell proliferation and migration, suggesting that the LSD1-SKP2 axis can serve as therapeutic targets for treatment of prostate cancers.

Citation Format: Liya Ding, Haojie Huang. LSD1 regulation of SKP2 expression in prostate cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1805. doi:10.1158/1538-7445.AM2013-1805
Abstract 1806: AP-1 transcriptionally regulates expression of miR-155 in colon cancer cells.
Authors: Onyeagucha Benjamin C. , Mercado-Pimentel Melania , Flemington Erik , Nelson Mark A.
Published: August 2014

Accumulating evidence indicates elevated S100P promotes the pathogenesis of cancers, including colon cancer. S100P exerts its effects by binding to and activating the Receptor for Advance Glycation End-products (RAGE). The effects of up-regulated S100P/RAGE signaling on cell functions are well documented. Despite these overwhelming evidences, little is known about the downstream targets of S100P/RAGE signaling. In the present study, we demonstrated for the first time that activation of RAGE by S100P regulates oncogenic microRNA-155 (miR-155) expression through Activator Protein-1 (AP-1) stimulation in colon cancer cells. Both S100P and miR-155 expressions are up-regulated in colon tumor specimens. Ectopic S100P expression leads to elevation of miR-155 level. Conversely, knockdown of S100P results in a decrease in miR-155 levels. Exogenous S100P induces miR-155 expression, but blockage of the RAGE receptor with anti-RAGE antibody suppresses the induction of miR-155 by exogenous S100P. Attenuation Blockage of AP-1 activation by S100P, through pharmacological inhibition of MEK activation or genetic inhibition of c-Jun activation using dominant negative c-Jun (TAM67) suppresses miR-155 induction by exogenous S100P. Exogenous S100P treatment stimulates the enrichment of c-Fos, an AP-1 family member at the miR-155 promoter site. Finally, functional study shows that miR-155 knockdown decreases colon cancer cell cell cell growtholon formation and motilityotility in S100P stably transfected cells and parental cells. Taken together, these data demonstrate that the expression of miR-155 is regulated by S100P is dependent on RAGE activation and stimulation of AP-1. Furthermore, the results show that miR-155 is a downstream target of S100P/RAGE signaling and a critical player in S100P functions in colon cancer cells.

Citation Format: Benjamin C. Onyeagucha, Melania Mercado-Pimentel, Erik Flemington, Mark A. Nelson. AP-1 transcriptionally regulates expression of miR-155 in colon cancer cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1806. doi:10.1158/1538-7445.AM2013-1806
Abstract 1830: Cancer-associated variants at 8q24 are correlated with expression of adjacent long non-coding RNAs involved in cell cycle regulation.
Authors: Kim Taewan , Croce Carlo M.
Published: August 2014

The mechanism by which cancer risk variants at 8q24.21 increase cancer risk is unclear for lack of protein-coding genes in the region, although such variants have been strongly associated with cancer development. By sequencing three colorectal cancer-associated variants and examining the expression levels of the lncRNAs at 8q24.21, we show that expression of long non-coding RNAs (lncRNAs) in the region is correlated with risk alleles of the cancer-associated variants. We further report that the lncRNAs regulate cell cycle by modulating expression of cell cycle-related genes such as CDKN1A and GADD45, suggesting the potential oncogenic functions of the lncRNAs. Our data provide new insight of disease-related variants in a gene desert.

Citation Format: Taewan Kim, Carlo M. Croce. Cancer-associated variants at 8q24 are correlated with expression of adjacent long non-coding RNAs involved in cell cycle regulation. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1830. doi:10.1158/1538-7445.AM2013-1830
Abstract 1832: Integrated analysis of clinical ChIPSeq data and public databases identified candidate long non-coding RNAs critically involved in gastrointestinal cancer.
Authors: Maruyama Reo , Yamamoto Eiichiro , Kumegawa Kohei , Tsuyada Akihiro , Ashida Masami , Suzuki Ryo , Tokino Takashi , Shinomura Yasuhisa , Suzuki Hiromu
Published: August 2014

Recent work has suggested various important functions and molecular mechanisms for long non-coding RNAs (lncRNAs) and they are also thought to play critical roles in cancer. However the importance of lncRNAs in cancer biology is still unclear. In this study, we sought to comprehensively identify novel lncRNAs that are critically involved in tumorigenesis and progression in gastrointestinal cancer. To identify candidate lncRNAs, we have applied the ChIP-Seq technology to clinical endoscopic biopsy specimens and characterized global histone modification patterns in more than 30 normal tissues and cancers. We developed an original lncRNA database based on several public databases. We then examined histone methylation status around putative lncRNAs and generated a list of candidate lncRNAs that may be important for cancer. In order to narrow the list down, we also utilized publicly available ChIPSeq data as well as bisulfite sequencing and RNASeq data to perform integrated analysis. Finally we identified ∼10 lncRNAs that are abnormally silenced by DNA methylation in gastrointestinal cancer and also identified other ∼10 lncRNAs that are abnormally overexpressed and potentially acting as oncogenic genes in cancer. We are examining gene expression level and epigenetic status of these lncRNAs in cancer cell lines and specimens by qRT-PCR, MSP, bisulfite pyro-sequencing and ChIP assay. Here we would like to show the advantage of making the best use of publicly available multi-omics data to promote long non-coding RNA research.

Citation Format: Reo Maruyama, Eiichiro Yamamoto, Kohei Kumegawa, Akihiro Tsuyada, Masami Ashida, Ryo Suzuki, Takashi Tokino, Yasuhisa Shinomura, Hiromu Suzuki. Integrated analysis of clinical ChIPSeq data and public databases identified candidate long non-coding RNAs critically involved in gastrointestinal cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1832. doi:10.1158/1538-7445.AM2013-1832
Abstract 1842: Identifying long noncoding RNAs in breast cancer microarrays.
Authors: Jia Hui , Lipovich Leonard , Ju Donghong , Kosir Mary A.
Published: August 2014

Long noncoding RNAs (lncRNA) are powerful regulators of gene expression. The emerging role of long noncoding RNAs (lncRNA) is facilitated by the development of new technologies such as tilling array and next generation sequencing. In this study, our research focused on lncRNA's regulatory mechanisms in breast cancer.

Methods: The Gencode v7 lncRNA and FANTOM5 lncRNA datasets were integrated as our lncRNA transcriptome. The expression profile was from four publications of microarray analysis in breast cancer. First, we checked the Affymetrix probe dataset U133 plus with our lncRNA transcriptome. If a probe was on the same strand and 100% fell into the exonic region of one lncRNA transcript, then the probe could this lncRNA. We integrated all lncRNAs from our transcriptome which matched the probes from the supplementary data of four papers and selected those occurred more than once.

Results: There were 23 nonredundant lncRNA candidates identified. Among those 23 candidates, PTENP1 was identified. PTENP1 is the pseudogene of tumor suppressor gene PTEN. The 3’UTR can bind the same miRNAs as the related protein-coding gene, PTEN. This suggests that pseudogenes may have a biological function as ‘decoys’, sequestering miRNAs and thereby affecting their regulation of expressed genes. Another candidate identified was ENST00000415285.1, which is the antisense gene of FGF14. FGF14 is a member of the fibroblast growth factor (FGF) family. FGF family members possess broad mitogenic and cell survival activities, and are involved in a variety of biological processes, including tumor growth and invasion.

ENST00000415285.1 is up-regulated in breast cancer, while the expression level of FGF14 is unknown, so further research is necessary to predict the possible mechanism of regulation to FGF14.

Conclusion: LncRNAs are identified in datasets for breast cancer that may elucidate further regulatory mechanisms driving the pathogenesis of breast cancer.

Citation Format: Hui Jia, Leonard Lipovich, Donghong Ju, Mary A. Kosir. Identifying long noncoding RNAs in breast cancer microarrays. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1842. doi:10.1158/1538-7445.AM2013-1842
Abstract 1861: Autophagy and cell death in breast cancer cells following oxidative stress by mitoquinone.
Authors: Gonzalez-Berrios Yanira , Aryal Baikuntha P. , Chehab Leena M. , Rao Ashutosh
Published: August 2014

Mitoquinone (MitoQ), a mitochondrially-targeted redox-active ubiquinone agent, selectively killed breast cancer cells compared to healthy cells mammary epithelial cells. While mitoquinone treatment led to irreversible loss in clonogenicity, the exact mechanism by which this agent was cytotoxic was unknown. We demonstrated that mitoquinone, although a synthetic derivative of the antioxidant ubiquinone, generated superoxide in MDA-MB-231 cells. Investigation into the cytotoxicity mechanism of MitoQ showed that a greater number of cells undergo autophagy than apoptosis. Drug treatment increased protein levels of the autophagosome-associated light chain 3 (LC3-II) that is responsible for the formation and nucleation of the autophagy bodies. Autophagy was confirmed by ultrastructural analysis using electron microscopy. However, the relationship between mitoquinone-induced oxidative stress and autophagy as a primary cellular response was not clear. We found that MitoQ induces Atg7-dependent but Beclin-1-independent cellular autophagy. Cancer cells and mouse embryonic fibroblast (MEF) lacking the autophagy related gene 7 (Atg7) had lower levels of autophagy after MitoQ treatment than Atg7 wild-type counterparts. In addition, we observed higher levels of oxidation in Atg7 deficient cells. Keap1, an oxidative stress sensor protein that regulates an antioxidant response via the transcription factor Nrf2, underwent oxidation and degradation in drug-treated cells. However, in Atg7 deficient cells Keap1 degradation decreased, suggesting Atg7-dependent Keap1 degradation. Conversely, knockdown of Keap1 decreased autophagy levels, but had no apparent effect on apoptosis. In vivo studies using spontaneously hypertensive rats implanted with a tumor confirmed that treatment with MitoQ reduced tumor size and increase autophagy in tumor cells. We established the anti-tumor activity of Mito-Q in breast cancer cell lines and in an animal model. We also provided preliminary evidence for an Atg7- and Keap1- dependent autophagy as one of the mechanisms that respond to the oxidative stress by Mito-Q.

Citation Format: Yanira Gonzalez-Berrios, Baikuntha P. Aryal, Leena M. Chehab, Ashutosh Rao. Autophagy and cell death in breast cancer cells following oxidative stress by mitoquinone. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1861. doi:10.1158/1538-7445.AM2013-1861
Abstract 1865: Ciglitazone increases ovarian cancer cell death by inhibiting GLUT-1 expression.
Authors: Kim Jin Young , Shin So Jin , Park Keon Uk , Jeon Young June , Cho Chi Heum , Ha Eunyoung
Published: August 2014

Ciglitazone is a kind of thiazolidinedione and a potent, selective peroxisome proliferator-activated receptor γ (PPAR γ) ligand and it is active in vivo as an anti-hyperglycemic agent in the murine model. Glucose metabolism plays a key role in the maintenance of energy homeostasis in organisms and changes in cellular glucose uptake rate are found in malignant diseases. Significant increase in glucose consumption can be found in many malignant conditions and research so far has focused on its apoptotic and inhibition of proliferation properties but their role in cell death is unclear. We show that ciglitazone inhibits glucose uptake in ovarian cancer cell through GLUT-1 expression. In this study, we investigated the effect of ciglitazone to GLUT-1 expression and cell proliferation. We further demonstrate that ciglitazone pretreated tumor cell was increased apoptosis dose dependent manner and siGLUT-1 treated tumor cell showed increased apoptosis. Interestingly, not only cancer cell line but also in vivo state, ciglitazone could decrease tumor mass.

Specificity protein (Sp) is highly expressed in tissues of prostate cancer, breast carcinomas and lung cancer when compared to normal tissues or cells. Previous studies showed that Sp1 plays an important role in carcinogenesis and metastasis of several human tumor types by regulating growth-related signal transduction, cell cycle control molecules, apoptosis, oncogenes, tumor suppressor genes. In this study there was no relation with apoptosis and SP1 expression. We demonstreated that ciglitazone inhibit glucose uptake in ovarian cancer cell lines and the relationship between ciglitazone and GLUT-1 in ciglitazone induced cell death.

Citation Format: Jin Young Kim, So Jin Shin, Keon Uk Park, Young June Jeon, Chi Heum Cho, Eunyoung Ha. Ciglitazone increases ovarian cancer cell death by inhibiting GLUT-1 expression. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1865. doi:10.1158/1538-7445.AM2013-1865
Abstract 1866: Stable isotope resolved metabolomics of colorectal cancer cells treated with inhibitor of monocarboxylate transporter-1.
Authors: Tan Bo , Kuo Ming-Shang , Breyer Matthew D. , Liu Ling , Zhao Yang
Published: August 2014

The rationale of this cell based study was to examine the metabolic effects of a synthetic monocarboxylate transporter 1 (MCT1) inhibitor (AR-C155858) on multiple cellular metabolites in colorectal tumor cells using [1,2,3,4,5,6-13C6]-D-glucose or [1,2,3,4,5-13C5]-D-glutamine as the single precursor metabolic tracer. DiFi colorectal cancer cells were cultured for 24 hrs in the presence of 0.1% DMSO (vehicle control) or 100 nM of MCT-1 inhibitor. Intracellular glycolysis intermediates measured by liquid chromatography/mass spectrometry (LC/MS) included phosphoenolpyruvate (PEP), phosphoglycerate (PG), fructose 1,6 bisphosphate (FBP), Glucose-6-phosphate (G6P), Gluconate-6-phosphate, glyceraldehydes 3-phosphate (G3P), ribose-5-phosphate (R5P), seduheptulose-7-phosphate (S7P), pyruvate, and lactate. TCA cycle intermediates measured included malate, α-ketoglutarate, succinate, fumarate, and citrate. Both glutamine and glutamate were also measured. Increases in PEP, PG3, G6P, Gluconate-6-phosphate, G3P, S7P, pyruvate, and lactate were induced by MCT1 inhibitor when traced with 13C6 glucose. When traced with 13C5 glutamine, PEP, G6P, S7P, pyruvate, and lactate were increased by MCT1 inhibitor. In contrast, decreases in α-ketoglutarate and glutamate were seen with both tracers.

Citation Format: Bo Tan, Ming-Shang Kuo, Matthew D. Breyer, Ling Liu, Yang Zhao. Stable isotope resolved metabolomics of colorectal cancer cells treated with inhibitor of monocarboxylate transporter-1. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1866. doi:10.1158/1538-7445.AM2013-1866
Abstract 1867: Metabolomic discrimination of undifferentiated and differentiated gastric cancer.
Authors: Kami Kenjiro , Fujimori Tamaki , Terashima Masanori , Kusuhara Masatoshi , Tokunaga Masanori , Tanizawa Yutaka , Bando Etsuro , Kawamura Taiichi , Yamamoto Hiroyuki , Ohashi Yoshiaki
Published: August 2014

Clinicopathological characteristics of gastric cancer patients strongly depend on histological type. In contrast with gene and protein expressions, metabolic properties of undifferentiated (UT) and differentiated (DT) gastric cancer have been largely unknown. Here, we conducted comparative metabolome analyses in UT and DT cell lines, uncovering characteristic differences between the levels of metabolites involved in energy metabolism. The metabolomic profile of DT cell lines was featured by its considerably high glycolytic intermediate, nucleotide, and glutathione levels, which highlight hallmarks of cancer metabolism represented by the Warburg effect and hyperactive nucleotide syntheses. In contrast, UT cell lines rather showed relatively low energy and redox statuses. We further confirmed the trends by measuring 392 metabolites of paired non-tumor and tumor gastric tissues obtained pairwise from 27 gastric cancer patients in order to metabolomically characterize non-tumors, UT (n=18), and DT (n=9) tumors. The results not only distinguished the profile of non-tumors from tumors, especially of DT, but also reaffirmed the distinctions between UT and DT tumors. Taken together, UT and DT cancers were metabolomically characterized both in vitro and in vivo; DT exhibited more typical cancer metabolic profiles than UT, whereas low glutathione ratios in UT imply their vulnerability to oxidative stress, which thus could be useful for developing more subtype-specific cancer therapeutics.

Citation Format: Kenjiro Kami, Tamaki Fujimori, Masanori Terashima, Masatoshi Kusuhara, Masanori Tokunaga, Yutaka Tanizawa, Etsuro Bando, Taiichi Kawamura, Hiroyuki Yamamoto, Yoshiaki Ohashi. Metabolomic discrimination of undifferentiated and differentiated gastric cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1867. doi:10.1158/1538-7445.AM2013-1867
Abstract 189: Preventive effects of branched-chain amino acids supplementation on the spontaneous development of hepatic preneoplastic lesions in C57BL/KsJ-db/db obese mice.
Authors: Shimizu Masahito , Terakura Daishi , Kochi Takahiro , Ohno Tomohiko , Kubota Masaya , Shirakami Yohei , Tanaka Takuji , Moriwaki Hisataka
Published: August 2014

Obesity and its associated disorders, such as non-alcoholic steatohepatitis, increase the risk of hepatocellular carcinoma (HCC). Branched-chain amino acids (BCAA), which improves protein malnutrition in patients with liver cirrhosis, reduces the risk of HCC in these patients with obesity. In the present study, the effects of BCAA supplementation on the spontaneous development of hepatic premalignant lesions, foci of cellular alteration (FCA), in db/db obese mice were examined. Male db/db mice were given a basal diet containing 3.0% of either BCAA or casein, a nitrogen content-matched control of BCAA, for 36 weeks. At sacrifice, supplementation with BCAA significantly inhibited the development of FCA when compared to casein supplementation by inhibiting cell proliferation, but inducing apoptosis. BCAA supplementation increased the expression levels of PPAR-γ, p21CIP1, and p27KIP1 mRNA and decreased the levels of c-fos and cyclin D1 mRNA in the liver. BCAA supplementation also reduced both the amount of hepatic triglyceride accumulation and the expression of IL-6, IL-1β, IL-18, and TNF-α mRNA in the liver. Increased macrophage infiltration was inhibited and the expression of IL-6, TNF-α, and MCP-1 mRNA in the white adipose tissue (WAT) were each decreased by BCAA supplementation. BCAA supplementation also reduced adipocyte size, while increasing the expression of PPAR-α, PPAR-γ, and adiponectin mRNA in the WAT compared to casein supplementation. These findings indicate that BCAA supplementation inhibits the early phase of obesity-related liver tumorigenesis by attenuating chronic inflammation in both the liver and WAT. BCAA supplementation may be useful in the chemoprevention of liver tumorigenesis in obese individuals.

Citation Format: Masahito Shimizu, Daishi Terakura, Takahiro Kochi, Tomohiko Ohno, Masaya Kubota, Yohei Shirakami, Takuji Tanaka, Hisataka Moriwaki. Preventive effects of branched-chain amino acids supplementation on the spontaneous development of hepatic preneoplastic lesions in C57BL/KsJ-db/db obese mice. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 189. doi:10.1158/1538-7445.AM2013-189
Abstract 1894: The lipogenic phenotype of HER2/neu-positive breast cancer cells.
Authors: Baumann Jan M. , Kourtidis Antonis , Conklin Douglas S.
Published: August 2014

15 - 30 % of all breast cancer cases are classified as HER2/neu-positive, a very aggressive form of breast cancer that correlates with a poor prognosis, reduced survival time and increased incidence of metastasis. Kourtidis et al. (2010) have recently shown that the HER2-positive breast cancer cell line BT474 relies on a unique Warburg-like metabolism for survival and aggressive behavior. These cells are dependent on fatty acid synthesis, presumably as a means to regenerate NAD+ for aerobic glycolysis. The nuclear receptors NR1D1 and PPARγ enable these cells to upregulate de novo fatty acid synthesis while avoiding palmitate-induced lipotoxicity by storing neutral fats (TAGs). The cells show markedly increased levels of stored fats compared to other breast cancer cell lines and disruption of the synthetic process results in apoptosis of BT474 cells. Metabolomic analysis of NR1D1 overexpressing cells compared to normal cells reveals profound differences in their metabolite profiles. This metabolic phenotype is operating at its limits in HER2-positive cells, as addition of exogenous palmitate results in cell death, whereas HER2-normal cells are not affected by this treatment. Microarray analysis suggests that palmitate induces an ER-stress response in the HER2-positive cells, ultimately resulting in cell death. Preliminary results suggest that this response is CHOP-dependent. Combination treatments with palmitate and HER2-targeted therapeutic compounds show synergistic effects and markedly increase the cytotoxic effects of either compound alone. The uptake of exogenous fatty acids as well as the de novo fatty acid synthesis are dependent on the essential co-factor coenzyme A (CoA), which is generated from the precursor pantothenate (vitamin B5). Based on our genomic analysis we predicted that HER2-positive breast cancer cells are more sensitive to changes in intracellular CoA levels. Our results show that HER2-positive breast cancer cells are sensitive to shRNA-mediated knockdown of coenzyme A pathway enzymes whereas MCF-7 cells are not. We observe the same pattern of sensitivity using the competitive CoA pathway inhibitor homopantothenate. Interestingly, limiting the supply of CoA in HER2-positive cells reduces the toxicity of exogenous palmitate.

Our data suggest that HER2-positive breast cancer cells are metabolically different from other breast cancer cell lines and that these differences offer new possibilities for therapeutic applications.

Citation Format: Jan M. Baumann, Antonis Kourtidis, Douglas S. Conklin. The lipogenic phenotype of HER2/neu-positive breast cancer cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1894. doi:10.1158/1538-7445.AM2013-1894