bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2518: Function of microRNA-10a in cholangiocarcinoma
Authors: Spriet Matthieu , Wehrkamp Cody , Henslee Brandon , Mohr Ashley , Kachman Steve , Mott Justin 
Published: September 2020

Abstract
Cholangiocarcinoma, a primary liver cancer of the bile ducts, effects 6,000 patients yearly and incidence has been growing consistently over the past 15 years. Current therapies for cholangiocarcinoma are not curative and patient survival is less than 1 year after diagnosis. Strides toward new treatment rest on better understanding tumor biology. The expression of microRNAs in cholangiocarcinoma cells was analyzed by RNAseq after knockdown of fibroblast growth factor receptor 4 (FGFR4) to identify potential microRNAs regulated by FGFR4. Caspase activity was used to evaluate miR-10a roles in anti-apoptotic signaling. Cell scratch assays were used to test cell migration and cell counting was used to determine proliferation. Quantitative real time PCR was used to measure miR-10a levels in cholangiocarcinoma cell lines Mz-ChA-1, HUCCT-1 and KMCH after FGFR4 knockdown with siRNA. Potential target proteins of miR-10a were evaluated by western blot. Preliminary data showed dozens of microRNAs (miRs) in cholangiocarcinoma cells were modulated by FGFR4 knockdown. Treatment with siRNA to FGFR4 caused increased sensitivity to TRAIL-induced apoptosis, and also reduced miR-10a expression in Mz-ChA-1, HuCCT-1, and KMCH cell lines. Importantly, apoptosis sensitization was reversed by restoring miR-10a expression in cells. miR-10a helped regulate both cell proliferation and cell migration in cholangiocarcinoma cells. When miR-10a was increased in cholangiocarcinoma cells, migration and proliferation increased. When miR-10a was decreased these effects also decreased. Protein targets of miR-10a included the MAP Kinase-related proteins MLK-1 & TAK-1. Other protein targets such as PHLDA1 & AKTIP were also identified. Notably, microRNA binding prediction programs did not include MLK-1 as a target, though our sequence analysis shows multiple stable binding sites. MLK-1 showed the most dramatic response to miR-10a. We investigated the role miR-10a plays for cholangiocarcinoma and the mechanism through which this pathway is regulated. Data supports miR-10a as a key player in cell survival, proliferation and migration. We have identified a set of proteins that are regulated by miR-10a in cholangiocarcinoma cells. Looking forward, identification of the function of these proteins will help elucidate how miR-10a functions in cholangiocarcinoma progression.
Citation Format: Matthieu Spriet, Cody Wehrkamp, Brandon Henslee, Ashley Mohr, Steve Kachman, Justin Mott. Function of microRNA-10a in cholangiocarcinoma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2518.
Abstract 2524: MiR-29b may suppress peritoneal metastasis via the effects on peritoneal mesothelial cells as well as on tumor cells
Authors: Kimura Yuki , Ohzawa Hideyuki , Saito Akira , Sata Naohiro , Kitayama Joji 
Published: September 2020

Abstract
Background: Peritoneal metastasis (PM) frequently occur in patients with gastric cancer with poor prognosis. Peritoneal cavity is lined with monolayer of mesothelial cells (MC) and contains various types of cells and exosomes. MC play an important role in the development of PM through mesothelial-mesenchymal transition (MMT). MiR-29 family is well known as tumor suppressor. However, the role of miR-29s in PM is not well characterized. We have previously reported that miR-29 family in exosome derived from peritoneal fluid was downregulated in patients with peritoneal metastasis of gastric cancer. In this study, we investigated the functions of miR-29s in the process of peritoneal metastasis.
Methods: Exosomes were purified from peritoneal fluids of 85 patients with gastric cancer and miR-29 levels in peritoneal exosomes were quantified with RT-PCR using TaqMan Advanced miRNA Assays. Proliferation and migration of human gastric cancer cell lines, MKN45 and NUGC, were examined with MTT and transwell assays, respectively. Human peritoneal mesothelial cells (HPMCs) were isolated by trypsinization of omental tissues obtained from patients undergoing bariatric surgery and cultured in DMEM supplemented with 20% fetal bovine serum. MiR-29b mimic and negative control were transfected by lipofection method using RNAiMAX. MMT was induced by the culture of HPMCs with 10 ng/ml TGF-β. Expressions of cytokeratin and vimentin were evaluated with flow cytometry.
Results: The expressions of miR-29a, miR-29b and miR-29c in peritoneal exosomes were all downregulated in 33 patients with PM as compared to those without PM. Transfection of miR-29b mimic significantly decreased the proliferation of MKN45 and NUGC-4 by 22~58% as compared with negative control miRNA (p&lt;0.01). Migration was inhibited more strongly by 84~90% (p&lt;0.01). After 48 hour-culture with TGF-β, HPMCs apparently changed the morphology from round to spindle shape. The expression level of cytokeratin in HPMCs was decreased while vimentin tended to be upregulated by TGF-β. However, transfection of miR-29b, but not control miRNA, significantly suppressed the morphological changes and reduced the expression levels of vimentin with restored expression of cytokeratin, suggesting the inhibition of MMT in HPMCs.
Conclusion: MiR-29b suppresses mesenchymal transition of MC as well as of tumor cells. Replacement of miR-29b to peritoneal cavity might be used for the treatment for peritoneal metastasis.
Citation Format: Yuki Kimura, Hideyuki Ohzawa, Akira Saito, Naohiro Sata, Joji Kitayama. MiR-29b may suppress peritoneal metastasis via the effects on peritoneal mesothelial cells as well as on tumor cells [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2524.
Abstract 2548: Pentose phosphate pathway inhibition potentiates metformin cytotoxic effects on HNSCC cells
Authors: Villaverde Marcela Solange , Gutkind Jorge Silvio 
Published: September 2020

Abstract
The pentose phosphate pathway (PPP) enables cancer cells to adapt to anabolic demands that require rapid DNA, RNA, and lipid synthesis and to oxidative cellular stress. Metformin can efficiently inhibit the malignant progression of oral premalignant lesions by indirectly inhibiting the mammalian target of rapamycin (mTORC1). Frequently activated in cancer cells, mTORC1 may upregulate the oxidative branch of the pentose phosphate pathway (PPP) through the sterol regulatory element binding protein (SREBP1). Our previous results showed that the combination of metformin and 6-AN (a PPP inhibitor) strongly potentiates their individual cytotoxic effects on melanoma, glioblastoma, mammary and colon carcinoma cell lines. However, the mechanism of this synergism remains unclear. In this context, the aim of this project was to determine the cell signaling events underlying the therapeutic potential of this combinatory approach, against HNSCC cell lines. Then, we study the effect of metformin alone or in combination with 6-AN on AMPK/mTOR/S6K/S6 and SREBP1 pathway. We found that the inhibition of PPP strongly potentiated metformin cytotoxic effect on 2D and 3D cultures of CAL33 and HN12, both HNSCC cell lines. Furthermore, the combination synergized to increase the activation of AMPK, concomitant with decreased phosphorylation of mTOR, decreased phosphorylation of S6K and decreased phosphorylation of S6. Interestingly, activation of SREBP1 was also synergistically decreased by this combinatory approach. We conclude that the inhibition of PPP and metformin converges to inhibit mTOR pathway. This interaction deserves to be further investigated as a potential strategy to potentiate metformin therapy.
Citation Format: Marcela Solange Villaverde, Jorge Silvio Gutkind. Pentose phosphate pathway inhibition potentiates metformin cytotoxic effects on HNSCC cells [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2548.
Abstract 2568: Characterization of metabolic changes induced by anti-cancer drugs in MCF7 and MDA-MB-231 cell lines
Authors: Hoffman David L. , Pisano Michael 
Published: September 2020

Abstract
Chemotherapeutics can have profound effects on cancer cell metabolism due to either direct or indirect effects. In many instances these effects can vary based on cell type, and level of cancer type aggressiveness. Here we focus on two breast cancer cell lines MCF7, and the triple negative MDA-MB-231 cells in a panel of assays designed to study specific aspects of cellular metabolism. The cells will be screened against three chemotherapeutics which have previously been shown to have effects on cellular metabolism, paclitaxel, AG-879 and oxaliplatin, and the mitochondrial targeted antioxidant mitoquinol (MitoQ), which has been shown to exhibit anticancer properties. Assays in this panel include cell growth, mitochondrial function, oxidative stress, lysosomal function and lipid storage. Through the utilization of this panel of assays panel, we compare the different metabolic phenotypes of two breast cancer cell lines with varied severity. Additionally, we demonstrate the effects that a small subset of anti-cancer agents has across a wide range of metabolic processes
Citation Format: David L. Hoffman, Michael Pisano. Characterization of metabolic changes induced by anti-cancer drugs in MCF7 and MDA-MB-231 cell lines [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2568.
Abstract 2579: Kras expression level regulates heparanase activity
Authors: Ju Donghong , Kosir Mary A. 
Published: September 2020

Abstract
Introduction: Sarcomas are locally and systemically aggressive, significantly interacting with extracellular matrix. The purpose of this study is to quantify the expression of heparanase by K-ras expressing sarcoma cells.
Methods: Balb/c3T3 and K234 (Ki-ras-transformed Balb/c 3T3) cells (ATCC) were cultured using DMEM with 5% FBS. Heparanase assays were performed with the heparan degrading enzyme assay kit (cat# MK412; Takara) on cell lysate for each cell line according manufacturer protocol. Dhmaracon on-Target Plus SMARTpool siRNA was used to knock down Kras. Total RNA was isolated with Qiagen RNeasy kit, cDNA was made using Invitrogen SuperScript™ III First-Strand Synthesis System, real time RT-PCR was done by use SYBR green Master Mix and Applied Biosystems® 7500 system, mRNA level for HPSE and PPBP was measured by RTPCR. The UCSC genome browser was used to show HPSE sequence and primer location.
Results: The heparan degrading activity of lysate from K234 cells was 20-30 times higher than from Balb/c3T3 cells. However, nine pairs of primers for mHPSE RT-PCR results show that mHPSE mRNA is expressed differently in Balb/c3T3 and K234 cell with K234 cell mHPSE mRNA level is lower than in Balb/c 3T3 cells. The mRNA sequence is the same in these two cell lines, however. Using siRNA knock down of greater than 80% Kras from K234 and Balb/c3T3 cells, the heparanase activity of lysate decreased by 78% in K234 cells, and 25% in Balb/c3T3 cells. Kras knock down also changed cell morphology in both cell lines, and changed the mRNA levels of HPSE, PPBP and some EMT markers.
Conclusion: Kras overexpressed in K234 cells is associated with increase in heparanase activity, and knock down of Kras decreases heparanase activity. However, heparanase activity does not correlate with HPSE (heparanase 1) expression level, so there is likely another mechanism(s) for Kras expression to regulate heparanase activity.
Citation Format: Donghong Ju, Mary A. Kosir. Kras expression level regulates heparanase activity [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2579.
Abstract 2580: Contrasting functions of EPSTI1 in human oral and lung squamous cell cancer cell lines - its tumor promoting roles in oral and tumor suppressing roles in lung squamous cell carcinomas
Authors: Fan Meng Meng , Arai Makoto , Tawada Akinobu , Chiba Tetsuhiro , Fukushima Reo , Uzawa Katsuhiro , Shiiba Masashi , Kato Naoya , Tanzawa Hideki , Takiguchi Yuichi 
Published: September 2020

Abstract
Introduction: Epithelial-stromal interaction 1 (EPSTI1) is a gene mapped to human chromosome 13q13.3. Recent studies have shown that EPSTI1 regulates many malignant features of cancers. However, the relevance of EPSTI1 to oral squamous cell carcinomas (OSCC) and lung squamous cell carcinoma (LSCC) are not known. The present study aimed to uncover the roles and the underlying mechanisms of EPSTI1 in OSCC and LSCC.
Methods: Quantitative real-time polymerase chain reaction (qRT-PCR) and western blot assay were used to evaluate the expression of EPSTI1 in four OSCC cell lines, HSC2, HSC3, HSC3M3 and HSC4, and three LSCC cell lines, LK-2, EBC-1 and H226. In vitro experiments were carried out in parental, their shRNA EPSTI1 knockdown and EPSTI1 overexpressing cells. Cell cycle was evaluated by flow cytometry, and epithelial-to-mesenchymal transition (EMT)-related proteins were evaluated by western blot assay. Changes in expression of EPSTI1-related genes in transfected cells were confirmed by using RT2 Profiler PCR Array Human Cancer Pathway Finder.
Results: EPSTI1 was significantly up-regulated in OSCC cell lines, and significantly down-regulated in LSCC cell lines compared to their normal counterparts. Knockdown of EPSTI1 in OSCC cells and overexpression of the gene in LSCC cells suppressed cell proliferation accompanied by cell-cycle arrest in the G1 phase with up-regulation of p21 and down-regulation of CDK2 and cyclin D1. These indicate that EPSTI1 was significantly involved in tumor growth in OSCC and LSCC, but in an opposite direction. Furthermore, EPSTI1 was related to enhanced cell migration and EMT phenotype in OSCC, and it was related to suppressed cell migration and reversed EMT in LSCC. In PCR-array analyses in two OSCC and two LSCC cells, some common genes were regulated similarly in each OSCC and LSCC.
Conclusion: EPSTI1 was up-regulated and its down-regulation suppressed the malignant phenotypes, indicating oncogenic roles of the gene in OSCC. In contrast the gene was down-regulated and its up-regulation suppressed the malignant phenotypes, indicating tumor suppressive roles of the gene in LSCC. These findings suggest that EPSTI1 might be a therapeutic target for OSCC and LSCC.
Citation Format: Meng Meng Fan, Makoto Arai, Akinobu Tawada, Tetsuhiro Chiba, Reo Fukushima, Katsuhiro Uzawa, Masashi Shiiba, Naoya Kato, Hideki Tanzawa, Yuichi Takiguchi. Contrasting functions of EPSTI1 in human oral and lung squamous cell cancer cell lines - its tumor promoting roles in oral and tumor suppressing roles in lung squamous cell carcinomas [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2580.
Abstract 2585: Tools for the discovery of KRAS pathway inhibitors
Authors: Kuznetsova Ekaterina , McCauley Lauren , Steinke-Neal Elaine , Sullivan Timothy , Primavera Louis , Ma Haiching 
Published: September 2020

Abstract
The small GTPase, KRAS, is a known oncogene that is frequently mutated in a large percentage of cancers and is associated with poor disease prognosis. Mutated KRAS is locked in the activated GTP bound state and facilitates enhanced Ras signaling in cancer cells. While being a desirable target, absence of good druggable binding pockets has made modulator compound discovery challenging and unsuccessful.
There has been a resurgence of interest in discovery of KRAS modulators after recent identification of unique binding pockets and successful inhibition of the KRAS G12C mutant by covalent chemical modifiers. With five molecules in clinical trials, many researchers are joining the efforts to find new inhibitors. Ras-mediated signaling commonly occurs through the receptor tyrosine kinase SOS-Ras pathway. To facilitate drug discovery activities targeting this pathway, we have produced the full spectrum of pathway proteins including kinases, wild type and mutated KRAS, and the upstream guanine nucleotide exchange factor protein SOS1. We have also developed and validated a number of biochemical and biophysical assays for these targets. This poster will summarize our most recent assay development efforts for the KRAS mutants including two different assays that monitor SOS1-mediated KRAS nucleotide exchange activities. Molecules that target SOS1 can be characterized using nucleotide exchange and KRAS::SOS1 protein-protein interaction assays. Binding of molecules to KRAS and SOS1 can also be assessed with biophysical methods. These tools and assays will significantly advance the drug discovery efforts of the industry.
Citation Format: Ekaterina Kuznetsova, Lauren McCauley, Elaine Steinke-Neal, Timothy Sullivan, Louis Primavera, Haiching Ma. Tools for the discovery of KRAS pathway inhibitors [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2585.
Abstract 2587: RAC1P29S expression in melanocytes is associated with increased DNA damage and altered cell cycle progression
Authors: Cannon Alexa C. , Chernoff Jonathan 
Published: September 2020

Abstract
Melanoma is the fifth most common cancer in the United States that has less than a 25% 5-year survival rate once the cancer has metastasized beyond the lymph nodes. Recent exome studies of cutaneous melanoma have identified RAC1P29S as the third most prevalent hotspot mutation in 4-9% of sun-exposed melanoma. RAC1 alteration in cancer is correlated to poor prognosis, resistance to standard chemotherapy, and insensitivity to targeted inhibitors. As a member of the Ras superfamily of small GTPases, RAC1 functions as a master regulator of actin cytoskeleton dynamics and influences proliferation, cell migration, motility, and polarity. Although RAC1P29S mutations in melanoma and RAC1 alterations in several other cancers are becoming increasingly evident, the RAC1-driven biological mechanisms contributing to melanomagenesis remain elusive. Lack of rigorous signaling analysis has prevented identification of alternative therapeutic targets for RAC1P29S melanomas. To investigate the RAC1P29S-driven effect on downstream molecular signaling pathways, we generated an inducible RAC1P29S expression melanocytic cell line and performed RNA-sequencing (RNA-seq) coupled with multiplexed kinase inhibitor beads and mass spectrometry (MIBs/MS) to establish enriched pathways from the genomic to proteomic level. Our proteogenomic analysis has identified the G2-M cell cycle phase transition as a hyperactive pathway in RAC1P29S-expressing melanocytes. RAC1P29S expression promotes increased kinase activity of the PAK-PLK1-AURORA A axis and increased transition into M-phase as indicated by FACS analysis. Interestingly, RAC1P29S-expressing melanocytes also possess increased DNA damage via γH2AX foci, suggesting a compromise in G2-M checkpoint integrity and an altered DNA damage response. Here, we assess the influence of RAC1P29S on the PAK-PLK1-AURORA A axis and its role in promoting G2-M transition. In addition, we investigate the DNA damage response and cell cycle regulation in the presence of cytotoxic agents. Collectively, these results suggest novel therapeutic pathways to target in RAC1P29S-driven melanoma.
Citation Format: Alexa C. Cannon, Jonathan Chernoff. RAC1P29S expression in melanocytes is associated with increased DNA damage and altered cell cycle progression [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2587.
Abstract 2593: Inhibition of COX1/2-driven thromboxane A2 pathway suppresses Barrett's esophagus and esophageal adenocarcinoma development
Authors: Zhang Tianshun , Wang Qiushi , Ma Wei-Ya , Wang Keke , Chang Xiaoyu , Johnson Michele L. , Bai Ruihua , Bode Ann M. , Foster Nathan R. , Falk Gary W. , Iyer Prasad G. , Dong Zigang 
Published: September 2020

Abstract
Barrett's esophagus (BE), a complication of gastroesophageal reflux disease (GERD), predisposes patients to esophageal adenocarcinoma (EAC). Reliable biomarkers for early detection and discovery of potential drug targets are urgently needed for improved BE and EAC patient outcomes. In this study, COX2 and TBXAS are highly expressed in BE and EAC patients accompanied by a pronounced elevation of circulating TXA2 levels in patient biopsy samples. Acetylsalicylic acid (ASA) suppressed BE and EAC growth by targeting the TXA2 pathway in a esophagoduodenostomy mouse model of reflux. Additionally, biopsies from patients showed that ASA substantially decreased serum TXA2 levels, resulting in reduced inflammation in a window opportunity clinical trial. Overall, this study establishes the importance of the COX1/2-driven TXA2 pathway in BE and EAC pathophysiology and lays the groundwork for introducing a TXA2-targeting strategy for EAC prevention and early detection. ASA targets the TXA2 pathway and suppresses BE and EAC.
Citation Format: Tianshun Zhang, Qiushi Wang, Wei-Ya Ma, Keke Wang, Xiaoyu Chang, Michele L. Johnson, Ruihua Bai, Ann M. Bode, Nathan R. Foster, Gary W. Falk, Prasad G. Iyer, Zigang Dong. Inhibition of COX1/2-driven thromboxane A2 pathway suppresses Barrett's esophagus and esophageal adenocarcinoma development [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2593.