bluuebunny/crossref_metadata_2025_split huggingface dataset:
2D vs 3D – Triple negative breast cancer spheroid formation induces quantitative heterogeneity of VEGF and PDGF receptor profiles and modulates cytosolic phosphorylation
Authors: Oyirifi Ashley , Joyce Kaitlyn M. , Helferich William G. , Imoukhuede Princess I 
Published: June 2021

Micro‐tumor spheres (MTS) have been considered a phenotypic intermediate between 2‐dimensional cell culture and tumor explants from living organisms. However, limited information exists for understanding membrane bound antigens of live cells in MTS. These antigens include several families of transmembrane proteins fundamental to signal transduction, cell communication, and adhesion processes and are thereby identified as key therapeutic targets in malignant cells. This study couples MTS cell culture and a high throughput method for live single cell receptoromic analysis, to compare MTS and monolayer cell surface densities of the PDGFRα, PDGFRβ, VEGFR1, and VEGFR2 tumor angiogenesis regulating receptors on MDB‐MB‐231.Cumulatively, the average density of PDGFRα, PDGFRβ, VEGFR1, and VEGFR2 in MTS is 1.7‐fold higher in tumor only MTS when compared to monolayer cells. When cells are analyzed on a cell‐by‐cell basis, 2‐component Gaussian mixture models predict means of high‐density and low‐density MTS sub‐populations within PDGFRα, PDGFRβ, VEGFR1, and VEGFR2 frequency distributions. The high density to low density means show 3.9‐, 1.6‐, 1.1‐, and 3.2‐fold differences for PDGFRα, PDGFRβ, VEGFR1, and VEGFR2, respectively. Finally, an analysis of MTS intracellular phosphorylation profiles preliminarily reveals attenuated phosphorylation, including reduced activation of the MAPK pathway, in MDA‐MB‐231 MTS simulated by a 20pg/mL VEGFA solution. Together, the current data suggests that MTS may provide a biologically relevant representation of breast cancer cells' VEGFR and PDGFR signaling axes through altering cell surface receptor density and heterogeneity. Our continued work will investigate the effects of varying VEGA concentrations on MTS signal transduction.Support or Funding InformationThis research was supported by NIEHS grant (T32 ES007326).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Decreased Expression of Enolase‐I Promotes Cisplatin Resistance of Ovarian Cancer Cells
Authors: Rabelo‐Fernández Robert Joe , Vivas‐Mejia Pablo , Santana‐Rivera Yasmarie 
Published: June 2021

Platinum‐containing anticancer drugs has been used for more than four decades has effective treatment for ovarian cancer and many others solid tumors. However, the initial positive response to therapy is limited by development of resistance to cisplatin treatment. Although many mechanism of cisplatin resistance have been proposed, the molecular mechanism leading to the cisplatin resistance in ovarian cancer tumors has not fully understood. In this study, we sought to identify novel proteins associated with the cisplatin resistance of ovarian cancer cells. Quantitative proteomic analysis revealed low protein levels of Enolase‐1 (ENO1) in cisplatin resistant (A2780CP20) ovarian cancer cells compared to its counterpart cisplatin sensitive (A2780) cells. Small‐interference RNA (siRNA)‐targeted ENO1 showed a reduction in the sensitivity of ovarian cancer cells to cisplatin treatment. Contrasting, ectopic expression of ENO1 in cisplatin resistant ovarian cancer cells reduced the sensitivity of these cells to cisplatin treatment. Intracellular glucose uptake were measurements in pairs of cisplatin sensitive and cisplatin resistant cells. The glucose content levels were higher in all cisplatin resistant cells compared with their sensitive counterparts. Overall, these results suggest that low levels of ENO1 are associated with the cisplatin resistance of ovarian cancer cells. Therefore, targeting the glycolytic pathways could be a strategy to overcome the cisplatin resistance of women with advanced and drug resistant ovarian cancer.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Pimavanserin tartrate: A potential drug for pancreatic cancer therapy in future
Authors: Ramachandran Sharavan , Srivastava Sanjay K 
Published: June 2021

Despite major advances in cancer treatment, pancreatic cancer is still incurable and the treatment outcomes are limited. In the current study, we evaluated the anti‐cancer effects of pimavanserin tartrate (PVT), a drug used for the treatment of Parkinson's disease psychosis. Our observations indicated that, PVT significantly suppressed the proliferation of pancreatic cancer cells by inducing apoptosis without exerting any cytotoxic effects in normal human pancreatic ductal epithelial (HPDE‐6) cells. The colony forming ability of pancreatic cancer cells was significantly inhibited with PVT treatment. Anti‐proliferative and apoptosis inducing effects of PVT were mediated by the inhibition of pAkt (Ser473), Akt, Gli1, Oct‐4, SOX‐2, NANOG and c‐Myc. Akt and Oct‐4 inhibition by PVT treatment was further validated by immunofluorescence analysis. Apoptotic effects of PVT was confirmed by increase in cleavage of caspase‐3 and PARP. Moreover, PVT inhibited the formation of tumorspheres in PANC1 pancreatic cancer cells. Pharmacologically inhibiting or genetically knocking out Akt or Gli1 enhanced the growth suppressive effects of PVT in pancreatic cancer cells. Subsequently, we evaluated the effects of PVT in gemcitabine‐resistant cells. Our results demonstrated that, PVT reduced the survival of MIAPaCa2 gemcitabine‐resistant cells in a concentration and time‐dependent manner. Further mechanistic analysis indicated that, PVT suppressed the phosphorylation of Akt at Ser 473 and inhibited the expression of Gli1, Oct‐4 and c‐Myc in MIAPaCa2 gemcitabine‐resistant cells. Moreover, PVT increased the cleavage of caspase‐3 and PARP as an indicator of apoptosis. Oral administration of PVT suppressed BxPC3 tumor xenografts by 50% in athymic nude mice. In another in vivo experiment, PVT treatment inhibited the growth of orthotopically implanted PANC1 tumors by 75%. Chronic administration of PVT did not exhibit any general signs of toxicity or behavioral side effects in mice. Taken together, our results indicate that pancreatic tumor growth suppression by PVT is orchestrated by inhibition of Akt/Gli1 signaling. Since PVT is already available in the clinic with an established safety profile, our results will accelerate its clinical development for the treatment of patients with pancreatic cancer.Support or Funding InformationSupported in part by R01 grant CA129038, awarded to (S.K.S.) by the National Cancer Institute, Syngenta fellowship grant awarded by Syngenta pharmaceuticals.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Endothelial RhoA Regulates Breast Cancer Metastasis
Authors: Sajib Md Sanaullah , Zahra Fatema Tuz , Park Jee Hyun , Tullar Paul , Srivastava Sanjay K. , Bickel Ulrich , Mikelis Constantinos M 
Published: June 2021

Metastasis is the cause of more than 90% of cancer‐related deaths. An important step in metastatic process is the migration of the disseminating cancer cells through the endothelial monolayer during entrance or exit from the vasculature. Activation of endothelial RhoA‐ROCK pathway has been implicated in the regulation of vascular permeability in various pathophysiological conditions. In the present study, we have investigated the role of this pathway in trans‐endothelial migration and metastasis of breast cancer cells. RhoA activation was identified by RhoA pull‐down experiments of murine and human breast cancer cell supernatant on primary endothelial cells. Cancer cell trans‐endothelial migration efficiency was explored through an in vitro transwell‐based two‐cell co‐culture model. RhoA signaling pathway was blocked by both pharmacological inhibition and knockdown experiments. In vivo metastatic potential was identified by experimental metastasis models of murine and human breast cancer cell lines using endothelial‐specific RhoA‐deficient mice or clinically relevant ROCK inhibitors. We identified that the diverse metastatic potential of a panel of human breast cancer cell lines correlated with the RhoA activation efficiency of their supernatant on primary endothelial cells. Secretome analysis revealed that IL‐8 (over)expression was relevant to breast cancer cell aggressiveness and its functionality was tested with gain‐and loss‐of‐function experiments. In vivo, experimental metastasis experiments with the syngeneic breast cancer cell line E0771 showed that endothelial RhoA deficiency decreased the metastatic potential. Similar inhibition was obtained with treatment with the clinically‐relevant ROCK inhibitor, Fasudil, in experimental metastasis experiments of murine and human cell lines using immune‐competent and immune‐deficient mice respectively. Collectively, our in vitro and in vivo findings demonstrate that the endothelial RhoA‐ROCK pathway affects breast cancer trans‐endothelial migration and metastasis and propose endothelial RhoA inhibition as a novel therapy for breast cancer metastases.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Protein Tyrosine Phosphatase Receptor Type F Promotes Wnt Signaling in Colorectal Cancer
Authors: Stevens Ashley , Gan Tong , Gao Tianyan 
Published: June 2021

Colorectal cancer is the second leading cause of cancer deaths in the United States. A major mechanism behind the formation of colon cancer is the activation of Wnt signaling through a mutation in the APC or β‐catenin gene, which leads to accumulation of active β‐catenin. Aberrant signaling of the Wnt pathway has been linked to the initiation and progression of colorectal cancer. A major mechanism of control for the Wnt signaling pathway is protein phosphorylation. A fine balance between phosphorylation and dephosphorylation exist to keep the cell balanced depending on its needs. Protein phosphatases are important in regulating signaling pathways by dephosphorylating a large number of cellular substrates. In this study, we investigated the functional importance of PTPRF, a member of protein tyrosine phosphatase receptor type 2A, in regulating Wnt signaling in colon cancer. Bioinformatics analysis indicated that PTPRF is highly expressed in the intestinal epithelium. Silencing PTPRF in colon cancer cells resulted in a decrease in the expression of Wnt target genes. As a consequence, the rate of cell proliferation was significantly decreased in PTPRF knockdown cells. To determine the molecular mechanism, we examined the internalization process of Wnt‐Fzd‐LRP6 receptor complex, a key mode of regulation for the Wnt pathway. Specifically, we monitored the time course of LRP6‐GFP tracking in control and PTPRF knockdown cells upon Wnt stimulation. Our preliminary confocal microscopy results showed that the amount of LRP6‐GFP protein detected at the cell surface was decreased basally and the time course of LRP6‐GFP endocytosis was altered upon Wnt treatment in PTPRF knockdown cells. Interestingly, we found that PTPRF was co‐localized with LRP6‐GFP at both endocytic vesicles and plasma membrane in colon cancer cells, and Wnt treatment promoted internalization of PTPRF. Taken together, our study has identified PTPRF as a novel regulator of Wnt signaling by controlling the endocytosis of LRP6 receptor in colorectal cancer.Support or Funding InformationThis work was supported by R01CA133429 and R01CA208343.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Investigating the Role of Inhibitors of NHE1 Activation as Potential Adjuvant Therapies in the Treatment of Ovarian Cancer
Authors: Corradi Anna R. , Wallert Mark A. 
Published: June 2021

Each year in the United States over 22,000 women are diagnosed with ovarian cancer leading to more than 14,000 deaths. Increases in the expression and activity Na+‐H+ Exchanger Isoform 1 (NHE1) was recently demonstrated to regulate cell proliferation in ovarian cancer patients. NHE1 is an 815 amino acid transmembrane transport protein that regulates intracellular pH. NHE1 is hyperactivated in an array of cancer types leading to a pH inversion that supports both cell proliferation and invasion. One mechanism for this NHE1 hyperactivation is phosphorylation of the regulatory domain by a range of protein kinases. A significant issue in ovarian cancer therapy is the development of drug resistance and disease recurrence. In an effort to combat these outcomes, it is hypothesized that combining common ovarian cancer therapeutic agents along with compounds that indirectly inhibit NHE1 activation will enhance the ability of the therapeutic agents to slow ovarian cancer progression. Two ovarian adenocarcinoma cell lines; CAOV‐3 and SKOV‐3 are used in this research. Using an XTT proliferation assay, the impact of different primary and second‐line ovarian cancer therapeutics alone and in combination with specific kinase inhibitors was evaluated. The therapeutics evaluated are: 1) Rucaparib, 2) Olaparib, 3) Paclitaxel, 4) Cisplatin, and 5) Carboplatin. An initial study of the protein kinases that regulate NHE1 activity was previously conducted. Four protein kinases that phosphorylate NHE1 at five specific locations: ROCK (RhoA‐Associated Kinase) at T653, RSK (Ribosomal S6 Kinase) at S703, AKT (Protein Kinase B) at S648, and ERK (Extracellular‐Signal Regulated Kinase) at S770/S771 were evaluated. The kinase inhibitors used were: MK220 (AKT), SCH772984 (ERK), Y2732 (ROCK), and BID1870 (RSK). Each of the pharmaceutical agents and the kinase inhibitors decreased cell proliferation independently in each of the cell lines. Proliferation data will be presented for the therapeutics independently and in the presence of kinase inhibitors. This work has also been expanded to a three‐dimensional environment, investigating the impact of these combination therapies in a spheroid assay. These data indicate that inhibition of NHE1 activating kinases may serve as a potential therapeutic target in Ovarian Cancer treatment.Support or Funding InformationLueken Family Cancer FundThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
ABC Transporter‐Mediated Multidrug Resistance in Prostate Cancer Cells
Authors: Famuyiwa Toluleke O 
Published: June 2021

BackgroundProstate cancer is the second most diagnosed cancer in men. The high incidence of prostate cancer has been attributed to a failure in chemotherapy. Studies show that energized ATP Binding Cassette proteins cause 50% of the failure in chemotherapy. This study focuses on inhibiting ATP Binding Cassette (ABC) protein‐mediated drug resistance in prostate cancer treatment by using Poly Lactic‐co‐Glycolic Acid (PLGA) nanoparticle to deliver SC‐514 and 3‐Bromopyruvate (3‐BPA).Objective of studyThis study aims to; (i) investigate the potential interaction between 3‐BPA and SC‐514, (ii) reduce treatment induced ABC mediated multidrug resistance (MDR), and (iii) investigate the signaling pathways involved in ABC transporter‐mediated MDR.MethodBioassays used in this preliminary study include trypan blue exclusion, MTT tetrazolium, NBT, cell titer glow, fluorescence microscopy, multidrug resistance efflux.ResultsOne‐way ANOVA comparing the impact of 3‐BPA, SC‐514 and the combination of 3‐BPA and SC‐514 on LNCaP cells at 24hrs of treatment shows that p=0.00023. Time‐dependent treatment of LNCaP cells with 3‐BPA, SC‐514, and a combination of 3‐BPA and SC‐514 after regression analysis shows the following p values: 24hrs (0.00023), 48hrs (0.00003), 72hrs (0.000000152), 96hrs (0.000000049). ROS level of LNCaP cells treated with 3‐BPA (r=−0.5, p=0.11), SC‐514 (, r=− 0.72 p=0.04,) and 3‐BPA + SC‐514 (r=−0.58, p=0.04) shows no significant difference in ROS modulation (p=0.54). There was a weak to moderate correlation between ROS released and cell death. There was a weak correlation between percentage ROS induced and percentage apoptotic death. There was a positive correlation between the concentration of the drug and cell death. 3‐BPA and/or SC‐514 depleted intracellular ATP in DU‐145 cells. There was a time‐dependent effect of 3‐BPA and/or SC‐514 on cell death. SC‐514 and/or 3‐BPA are substrates for MDR1. 3‐BPA and/or SC‐514 potentially block MDR1. DU‐145 prostate cancer cells efflux dyes within 15 minutes.Conclusion3‐BPA and SC‐514 has the potential to inhibit multidrug resistance by reducing the intracellular ATP available to ATP binding cassette proteins. Apoptotic induction in DU‐145 prostate cancer cells exists via a mechanism other than reactive oxygen species (ROS) induction.Support or Funding InformationFellowships and grantsThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Investigation of Antimicrobial Properties of Essential and Natural Oils on Cronobacter Species
Authors: Whitehead Nickolas T. , Kanipes Margaret I. , Fakayode Sayo O. 
Published: June 2021

The use of essential and natural oils are becoming increasingly popular due to their antimicrobial, therapeutic antioxidant properties. Research has shown that essential and natural oils are effective against a variety of bacteria. However, not much experimentation has been conducted to investigate the antibacterial properties of essential oils and natural oils on Cronobacter species. This study evaluates the antibacterial property of ten essential and natural oil samples on Cronobacter species using a modified disk diffusion assay using a ciprofloxacin as control. The results of the study shows differences in the antibacterial properties of essential and natural oils. The essential oils that inhibited cell growth of Cronobacter muytjensii were lemon eucalyptus oil (Eucalyptus citriadora), lemon oil (Citrus limonum), tea tree lemon oil (Leptospermum petersonii), and wintergreen oil (Gaultheria fragrantissima). Neem oil (Azadirachta indica) (a natural oil), displayed signs of reduced growth in the bacteria. The inhibitory properties of the aforementioned oils will be discussed.Support or Funding InformationNorth Carolina A&T University Research Initiative for Scientific Enhancement (RiSE) ProgramThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Hypoglycemia effect of drink containing Guava Leaf Extract in Rats with type 2 diabetic rats
Authors: Huang Hui‐Yu , Chen Li‐Han , Chan Ching‐Hung 
Published: June 2021

This study investigated the effect of drink containing Guava Leaf Extract (GLE) (Psidium guajava Linn.) on hypoglycemia and glucose metabolism in type 2 diabetic rats. High fat‐fructose Diet and Low‐dose streptozotocin (STZ) and nicotinamide were given and injected into Sprague‐Dawley (SD) rats to induce type 2 diabetes. Fifty rats were equally assigned into diabetes mellitus group (DM), positive control group (PC), low‐dose GLE group (L), medium‐dose GLE group (M), and high‐dose GLE group (H) respectively. We found that GLE progressively decreased fasting blood glucose and OGTT (□ < 0.01). Elevated pro‐inflammatory cytokines, interleukin‐1β (IL‐1β), IL‐6, and tumor necrosis factor‐α (TNF‐α), were controlled by L, M and H group. Antioxidant activity was increased in L, M and H group. Experimental evidences demonstrate that GLE may be effective alternative medicine to prevent HFFD‐induced hyperglycemia and hyperlipidemia, possibly associated with anti‐inflammatory and antioxidant efficacies.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Investigating a Novel Metabolic Pathway for Bacteria to Utilize L‐Ascorbate as its Carbon Source
Authors: Kim Peter , Stack Tyler , Carter Michael 
Published: June 2021

L‐Ascorbate (Vitamin C) is an abundant yet under‐researched carbon source present in the environment. L‐Ascorbate is a well‐known antioxidant that has significant effects of promoting tissue repair, strengthening bones, and boosting the immune system in mammals. Ralstonia eutropha H16 (Cupriavidus necator) is a saprotrophic soil bacterium that is commonly known for its ability to metabolize a great variety and number of substrates for growth and production of polyhydroxybutyrate (bioplastics). Ralstonia eutropha (ReH16) growth with L‐ascorbate was observed, but the pathway by which ReH16 metabolizes L‐ascorbate is unknown. Transcripts were quantified using RNAseq from cells grown with L‐ascorbate or other carbon sources. Putative pathway genes were identified as genes with transcripts upregulated greater than 50‐fold during L‐ascorbate growth. Transformation, conjugation, homologous recombination, and PCR analysis were used to generate and confirm targeted deletions of two pathway genes [h16_B0212 and h16_B1217]. Mutants of both genes were unable to grow with L‐ascorbate as their sole carbon source. Collaborators have characterized both enzymes. H16_B0212 is an aldehyde dehydrogenase that catalyzes oxidation of the δ‐carbon of α‐ketoglutarate semialdehyde. H16_B1217 is an enzyme that catalyzes the formation of a β‐lactone ring and transfer of a carboxyl group. Characterizing the L‐ascorbate metabolic pathway in Ralsontia eutropha will provide insight into a third, yet unknown pathway for L‐ascorbate metabolism.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Tempol, a superoxide dismutase mimetic agent, improves dystrophic phenotype in the diaphragm muscle of mdx mice
Authors: Minatel Elaine , de Almeida Hermes Túlio , Mâncio Rafael Dias , Macedo Aline Barbosa , Mizobuti Daniela Sayuri , da Rocha Guilherme Luiz 
Published: June 2021

Duchenne muscular dystrophy (DMD) is a progressive and lethal X‐linked myopathy, in which the oxidative stress and abnormal reactive oxygen species (ROS) production plays a key role in its pathophysiology. Despite intense efforts, no cure is currently available for DMD and glucocorticoids are the only effective drug to slow down disease progression. Thus, in the present study we investigated Tempol (a superoxide dismutase mimetic agent) effects on the diaphragm muscle of mdx mice, an experimental model of DMD. Mdx mice were randomly divided into two groups: mdxS, the control group receiving intraperitoneal (i.p.) injections of saline solution (100μL) and mdxT, the treated group receiving i.p. injections of Tempol (100 mg/kg). C57BL/10 mice were also used as controls. The results showed that Tempol treatment prevented muscle damage indicated by gain in muscle strength, decrease in the IgG uptake by muscle fibers and reduction of regenerated muscle fibers indicated by the central nuclei. It also had an effect on the inflammatory process, reducing the inflammatory area, macrophage staining and decreasing TNF‐α, IL‐1β and IL‐6 levels. Regarding oxidative stress, Tempol treatment was effective in reducing ROS and lipid peroxidation in the dystrophic diaphragm muscle. The findings presented here show that Tempol treatment improves dystrophic phenotype, supporting its use as a potential therapeutic strategy in DMD.Support or Funding InformationFAPESP #2017/01638‐0CAPES FAEPEXThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.