bluuebunny/crossref_metadata_2025_split huggingface dataset:
SOD2 confers development of resistance to PI3K pathway inhibition associated to enhanced stemness features in head and neck cancer
Authors: Chang Kwang‐Yu , Hsueh Wei‐Ting , Chu Jui‐Mei , Chou Shao‐Wen , Chang Wen‐Chang , Chuang Jian‐Ying 
Published: June 2021

BackgroundRecent development of targeting therapies in head and neck cancer (HNC) focuses on the phosphoinositide 3‐kinase (PI3K) pathway, which is known to be a crucial factor in cancer development and resistance to multiple drugs. The effect of the specific inhibitors, however, is often overshadowed by the development of resistance which the mechanism remains to be determined.MethodBEZ235, a dual inhibitor targeting against PI3K and the mammalian target of rapamycin (mTOR), was used to develop resistant HNC lines by long‐term co‐incubation and selection of the single cell clones. Cellular profiles of reactive oxidative stress (ROS) and the stemness features were explored to analyze the PI3K related functions.ResultsThe resistant cells showed higher IC50 of the proliferation in presence of the PI3K/mTOR dual inhibitors BEZ235 and BGT226, the specific PI3K inhibitor LY294002, and the mTOR inhibitor rapamycin. They exhibited cross resistance to an EGFR inhibitor gefitinib and a platium chemotherapy cisplatin. Examination of the signaling in presence of BEZ235 showed persistent PI3K activation with expression of phosphorylated AKT at threonine 308 in the resistant cells. The activated AKT signaling was also noted when the cells were treated with cisplatin. the stemness features were assessed by flow cytometry with increased ALDH level noted in the resistant cells. This was supported by increased associated biomarkers in qPCR and higher Bmi1, Sox2 expression in Western blot. Functional studies indicated increased expression of ROS in the resistant cell lines. Analysis of the ROS scavengers revealed higher SOD2 mRNA level by qPCR but not SOD1. Notably from a publicly available cancer microarray database Oncomine, higher SOD2 expression was predominantly shown comparing to the other scavengers in the HNC tissues than the normal ones. Finally, the association of SOD2 in the stemness‐featured cells were shown by higher gene expression in the Alderfluor‐stained ALDH‐positive parental cells. Since treatment with BEZ235 further augmented SOD2 expression, co‐incubation of SOD inhibitors and BEZ235 reduced development of spheroid cells and showed advantage in successful inhibition of these stemness‐featured cells.ConclusionAcquisition of resistance may involve alterations of cell population which has more dominant survival mechanism. This may contribute to the cross‐resistance of the other anti‐tumor drug. Intracellular ROS modulation has crucial role in the process. Early intervention may be needed for keeping cells from developing resistance.Support or Funding InformationThis study was supported by the grant from National Health Research Institutes, Taiwan (CA‐107‐PP‐08)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Primary cilia in cancer cells
Authors: Jamal Maha Hassan , Nauli Surya 
Published: June 2021

Primary cilia are sensory organelles that exist on the surface of most cells. Since primary cilia play an important role in cell cycle and cilia‐related signaling pathways (e.g. Hedghog and Wnt signaling pathways), ciliary dysfunction has been proposed as an essential factor of cancer development. In our current study, we characterized the expression of the primary cilia in two breast cancer (MCF7 and MDA132), two prostate cancer (PC3 and DU145), and one lung cancer (NL20‐TA) cell lines by counting number of cilia formation and measuring cilia length. We also examined the effect of shear stress on the cilia number and cilia length. Moreover, we investigated the correlation between primary cilia and cilia‐related signaling pathways. We found that the frequency and the length of the primary cilia were reduced in MCF7 and MDA‐MB‐231 breast cancer cells and in PC3 prostate cancer. Primary cilia were absent in DU‐145 prostate cancer and NL20‐TA lung cancer. The β‐catenin of Wnt signaling pathway was elevated in the prostate and lung cancers, while Gli1 and smoothened of Hedghog pathway remained the same. Our results suggested an association between primary cilia and Wnt signaling in cancer cells.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Bisdemethoxycurcumin potentiates gemcitabine to induce mitochondrial dysfunctions and apoptosis in human pancreatic adenocarcinoma cells
Authors: Li Xuejun 
Published: June 2021

Pancreatic cancer is a highly aggressive malignancy, gemcitabine is indicated as a standard single agent for the treatment of pancreatic cancer, however it only extends survival by a mere 3–6 weeks. In this present study we investigate whether bisdemethoxycurcumin (BDMC), a derivative of curcumin, potentiates gemcitabine in human pancreatic cancer cells. The result suggested that BDMC produced a synergistic effects with gemcitabine by inducing mitochondrial dysfunctions and apoptosis in PANC‐1 and MiaPaCa‐2 pancreatic cancer cells. By using 2D‐MS, we found 13 essential proteins were significantly changed in response to gemcitabine alone or combined with BDMC. Pharmacological network analysis revealed that glucose‐regulated protein 78 (GRP78) as the hub activated by BDMC. We then demonstrated that BDMC upregulates GRP78 and facilitated apoptosis through eIF2α/CHOP pathway. In addition, DJ‐1 and prohibitin, two identified markers of chemoresistance, were increased by gemcitabine in PANC‐1 cells. This could be reversed by BDMC, suggesting that BDMC partially offsets the chemoresistance induced by gemcitabine. In conclusion, our study shows that BDMC promotes apoptosis through a GRP78‐dependent pathway and mitochondrial dysfunctions, and potentiates the antitumor effect of gemcitabine in human pancreatic cancer cells.Support or Funding InformationThis work was supported by the National Natural Science Foundation of China (No. 81874318, 81673453, 81473235).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inhibitory effects of caffeic acid phenethyl ester (CAPE) derivatives on multiple myeloma cell growth
Authors: Jackson My Lien , Wang Xinyu 
Published: June 2021

Multiple myeloma (MM) is a rare blood cancer that affects thousands of people yearly in the U.S. It stems from monoclonal plasma cells and can cause symptoms such as bone loss and fatigue. Recent studies have shown that the bone marrow microenvironment may mediate the tumor proliferation, drug resistance, and migration of the MM cells. Caffeic acid phenethyl ester (CAPE) has been studied in research due to their anti‐cancer and anti‐inflammatory effects. The objective of this study is to examine the inhibitory effects of CAPE derivatives on MM cell growth and explore the structure‐activity relationship of CAPE. Selected CAPE derivatives include ferulic acid methyl ester (FAME), caffeic acid methyl ester (CAME), caffeic acid (CA), N‐octyl caffeate (NOC), phenethyl dimethyl caffeate (PDC), phenylethyl‐4‐methylcaffeate (P4MC), and fluorinated CAPE (FCAPE). These seven CAPE derivatives with a dose range from 1.0 to 50 μM were tested on the growth of MM RPMI 8226 cells within 72 hours. Our current results show that FCAPE, P4MC, and NOC decreased cell viability and proliferation of the MM 8226 cells in a dose and time‐dependent manner. These derivatives were selected to further study whether its inhibitory effect on MM cell growth is due to the induction of apoptosis. Western blotting was used to evaluate the protein expression of apoptosis‐related factors such as caspase 3, caspase 8, caspase 9 and poly‐(ADP‐ribose) polymerase (PARP). To evaluate the effect of these CAPE derivatives on MM 8226 cells in the presence of the microenvironment, a Transwell (TW) model using MM cells co‐culturing with bone marrow stromal cells will be applied to mimic the bone marrow microenvironment in vivo. When treating the HS‐5 stromal cells with CAPE derivatives, no significant decrease in proliferation is seen after 24 hours. We expect that after co‐culturing the MM 8226 cells with HS‐5 stromal cells using the TW model, the growth of MM cells will still be prohibited by those CAPE derivatives to some extent if not the same as MM cells being treated alone. In conclusion, our data suggest that CAPE derivatives inhibit the growth of MM cells through induction of apoptosis, and this effect of CAPE requires the integrity of a catechol ring functional group in order to carry out its cytotoxic effects.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Ethylacetate fractional extract of Mentha spicata attenuates irinotecan‐induced mucositis in mice
Authors: Kathem Sarmed H , Abdu‐Jabbar Aliaa A , Ataimish Ahmed H , zalzala Munaf H 
Published: June 2021

Irinotecan induced‐mucositis is an inflammatory event of intestine that has negative impact on patient health and affect treatment efficacy. Irinotecan first metabolized in the liver by a carboxylesterase (CES) to the active cytotoxic metabolite (SN‐38), which subsequently glucuronidated by hepatic enzymes to the inactive metabolite SN38G. The SN‐38G undergo deconjugation in the intestine liberating the active SN‐38 via β‐glucuronidase produced by the intestinal bacterial flora. Consequently, SN‐38 triggers inflammation events characterized by delayed intestinal mucositis and diarrhea. This study aims to ameliorate irinotecan‐induced mucositis and diarrhea in mice model using ethylacetate fractional extraction of Mentha spicata(EAEMS), an edible widley consumed plant. Mice model of irinotecan‐induced mucositis and diarrhea produced by I.P injection of irinotecan (75mg/Kg/day) for 4 days. EAEMS administered orally (160mg/kg) twice daily for 7 days starting one day before irinite can dose. Diarrhae scores and mice body weight variation measured to assess the severity of mucositis and diarrhea. Jejunal IL‐1β and MPO measured as markers of intestinal inflammation. β‐glucuronidase activity in fecal pelletes were measured also. Compared to model control mice, results revealed that animals received EAEMS presented with significantly lower diarrhea scores and body weight loss. Furthermore, jejunal IL‐1β and MPO significantly reduced and associated with significant inhibition of fecal β‐glucuronidase activity in animals received EAEMS compared to model control mice. In conclusion, we showed for the first time that EAEMS has a protective effect against irinotecan‐induced mucositis and diarrhea, an effect suggested to be mediated by inhibition of bacterial β‐glucuronidase activity. This promising result might have a therapeutic application in cancer patients suffering from delayed‐onset diarrhea due to irinotecan treatment.Support or Funding InformationUniversity of Baghdad, College of PharmacyThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Anticancer Activity of Aqueous Extracts of Terminalia arjuna (TA) Bark
Authors: Liu Shi J , Liu Meeiyueh , Li Hong‐yu J , Simmen Rosalia CM , Johann Donald J 
Published: June 2021

Our previous study demonstrated that an aqueous extract of TA bark (TAAqE) exerts cardioprotection against toxicity induced by doxorubicin (DOX), a commonly‐used anticancer chemotherapeutic agent. However, whether TAAqE has any effect on cancer cells remains unknown. Herein we investigated the effects of TAAqE on several human lung and breast cancer cell lines (ATCC) in vitro and on mice (Wnt‐transgenic model) bearing mammary tumors. Cell morphology and growth were monitored with phase‐contrast microscopy, while the colorimetric Cell Counting Kit (CCK)‐8 assay was used to evaluate cell viability. Fluorescent microscopy with Hoechst H33342, AnnexinV, propidium iodide (PI), and JC‐1 (a mitochondrial membrane potential dye) was used to detect cell membrane and mitochondrial integrity. Tumor growth in vivo was monitored with ultrasound biomicroscopy. We found that TAAqE, its fractions (designated as P faction and S fraction) and the ethanol extract of TA bark (TAEtOH) at 50 μg/ml inhibited growth of all cancer cells with negligible effect on non‐tumor lung fibroblasts (CCD‐19Lu) after 24‐h treatment. The order of the cytotoxic potency (at 50 μg/ml) was TAEtOH > P fraction > TAAqE ~ S fraction > gallic acid (10 μM), an active component of TAAqE, based on the cell viability assay. When TAAqE was separated with Microcon filter devices, the molecular mass of active components was greater than 10 kDa. Distinct AnnexinV/PI expression of these cancer cells suggested differential effects of TAAqE on apoptotic status. TAAqE and TAEtOH caused dose‐dependent cytotoxicity of human lung adenocarcinoma cells (A549) with 50 μg/ml showing more effective effects and different morphological changes such as increasing cytosolic vacuolization compared to DOX (1 μM). In non‐small cell lung cancer cells (H1975), TAAqE (5 μg/ml) caused a decrease in mitochondrial membrane potential in contrast to CCD‐19LU cells. In vivo treatment of Wnt‐transgenic mice with TAAqE (15 mg/kg body weight, daily in drinking water) for 3 days or 4 weeks reduced mammary tumor sizes with no recurrence noted 3.5 weeks and one month post‐treatment, respectively. In conclusion, TAAqE displays potential as an adjunct for cancer therapy in addition to its cardiotoxic protective effects. Mechanistically, TAAqE may induce cell membrane damage and reduction of mitochondrial function of cancer cells.Support or Funding InformationUAMS Seeds of Science Funds to SJLThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Induction of rat models with myocardial ischemia or anoxia and the expression of succinate receptor
Authors: Liu Yongxue , Zhang Fangyuan , Qi Yue , Yuan Meiru 
Published: June 2021

ObjectiveTo establish rat models with myocardial ischemia and anoxia and to detect the expression of succinate receptor (GPR91) in rat myocardium.MethodsTotal 48 Sprague‐Dawley (SD) rats used in the experiment. For the induction of myocardial ischemia, eighteen rats were randomly divided into three groups of sham operation (control), coronary artery ligation 12h and 24h, respectively. The rats were anaesthetized with sodium pentobarbital and ligated by the left anterior descending artery for 12h and 24h. The control rats were operated as the same except ligation. The electrocardiogram (ECG), the area of myocardial infarction and the serum contents of lactate dehydrogenase (LDH), creatine kinase (CK) and cardiac troponin 1 (cTn‐1) in rats were recorded and detected at the setting time. For the induction of myocardial anoxia, thirty rats were divided into five groups of living in normal atmosphere (control), living in low atmosphere for 1d, 3d, 5d, and 7d. At the setting days, the serum contents of lactate LDH, CK and cTn‐1 in rats were detected. Real‐time fluorescent quantitative PCR and Western Blot were used for GPR91 mRNA and protein expression in rat myocardium.ResultsIn the rats ligated by the left anterior descending artery for 12h and 24h, continuous ST segment elevations were recorded in ECG; the ratio of infarct size were 23.18±3.46 and 34.85±2.22 (P<0.05, compared with the control); the serum levels of LDH, CK and cTn‐1 were increased in some extent. In the rats with myocardial anoxia for 1d, 3 d, 5d, and 7d, elevation of LDH, CK and cTn‐1 contents in serum were also detected. Compared with that of the control rats, the expression of GPR91 mRNA and protein in myocardium of myocardial ischemia or anoxia rats were both up‐regulated.ConclusionThe left anterior descending artery ligation or living in low atmosphere could induce myocardial ischemia or anoxia in rats. The expression of GPR91 mRNA and protein in myocardium of rats with myocardial ischemia or anoxia was increased. It implied that GPR91 could serve as a target for prevention and treatment of ischemic or anoxia myocardial injury.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Novel Mechanism of Regulation of Sodium Channel Nav1.5 Expression in Hypoxic Cardiomyocytes
Authors: Wang Ning , Liu Yining , Li Jiamin , Yang Zhenyu , Cai Benzhi , Xu Haodong 
Published: June 2021

HypothesisBecause of the increased susceptibility of arrhythmias in patients with myocardial infarction (MI) in clinic, we hypothesized that Meis1, a critical developmental transcriptional factor, regulates NaV1.5 expression by affecting the SCN5a promoter activity, which leads to an alteration of cardiac electrical conduction and has a potential role in the regulation of NaV1.5 expression in hypoxic cardiomyocytes.MethodsMI mouse model was induced by ligation of the left anterior descending artery (LAD) of mouse heart. Echocardiography and electrocardiogram (ECG) were performed to determine cardiac function and electrical activity, respectively. qRT‐PCR and western blot were used to determine the NaV1.5 and Meis1 expression levels in peri‐infarct (border zone) region of MI mouse hearts. In vitro, neonatal mouse ventricular cardiomyocytes were treatment with hypoxia (1% O2, 5% CO2, 94% N2) for 24 hours and further infected with Adeno‐Associated Viral‐Meis1 (AVV‐Meis1) and transfected with siRNA specifically targeting Meis1 (si‐Meis1). qRT‐PCR and Western blot were employed to determine if increased or decreased Meis1 had effects on NaV1.5 expression. In silico promoter analysis was performed to determine if the SCN5a promoter harbors Meis1 conservative binding sites in different species. Luciferase activity assay was performed to observe the influence of Meis1 on the SCN5a promoter activity in HeLa cells.ResultsSignificantly prolongation of QTc interval was induced in mouse with MI operation. Both mRNA and protein levels of Meis1 and NaV1.5 were significantly decreased in infarcted hearts of the mice, compared to the sham group. In vitro, both mRNA and protein levels of NaV1.5 and Meis1 were significantly decreased in hypoxic ventricular cardiomyocytes of neonatal mouse, in comparison with a control group. Overexpression or knockdown of Meis1 was shown to enhance or reduce the expression of NaV1.5 mRNA and protein in ventricular cardiomyocytes of neonatal mouse. In silico promoter analysis revealed that the SCN5a promoter contains Meis1 conservative binding DNA sequences in humans, mice and rats. Luciferase reporter assay preformed with HeLa cells showed that Meis1 overexpression increased the SCN5a promoter activity.ConclusionCollectively, downregulation of Meis1 significantly contributes to NaV1.5 reduction after MI. The underlying molecular mechanism is that meis1 transcriptionally regulates the expression of the NaV1.5 protein by increasing SCN5a promoter activity. Our findings provide a new insight into the molecular mechanisms of sodium channel NaV1.5 expression in the development of ventricular arrhythmias related to MI.Support or Funding InformationThe study was supported by the National Nature Science Foundation of China (grants 81573425 and 81773733).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Deficiency of CD4 T cells Contributes to Cardiac Fibrosis and Myocardial Dysfunction in T cell Specific S1P Receptor 1 Knockout Mice
Authors: Jin Zhuqiu , Abdullah Chowdhury 
Published: June 2021

Sphingosine 1‐phosphate (S1P) is a bioactive small lipid molecule involved in apoptosis, barrier formation and function, cytoskeleton rearrangement, vascular maturation, wound healing, cell trafficking, and other functions via five G protein‐coupled S1P receptors or/and intracellular lipid signaling. T cell S1P receptor 1 (S1P1) is responsible for mature T cell exit from thymus to the blood and periphery. Nevertheless, effects of T lymphocyte S1P1 on cardiac function and fibrosis remains incomplete understood. To this purpose, we generated T cell S1P1 knockout mice (TS1P1KO) by crossing S1P1loxP/loxP mice with Lck‐Cre mice. Body weight (g) and glucose levels (mg/dl) in the blood were monitored. CD4 and CD8 T lymphocytes were counted by using flow cytometry. Pathological grading of heart histology was conducted in H&E stained sections. Masson Trichrome staining was utilized to evaluate cardiac fibrosis. Cardiac contractility was determined in Langendorff perfusion system. The TS1P1KO mice exhibited sustained deficiency of both CD4 and CD8 T cells in the blood compared with wild‐type littermate control. The TS1P1KO mouse heart featured an altered phenotype characterized by reduced cardiac contractility and increased cardiac fibrosis as well as morphological alteration, notably, loss of interstitial space between cardiac myocytes with thickening of vascular smooth muscle cells and perivascular edema compared with littermate vehicle mice. To explore the role of CD4 T cells in altered phenotype, naïve CD4 T cells were isolated by positive selection through MS column from CD4 magnetic microbeads‐labeled wild‐type littermate mouse splenocytes. Isolated CD4 T cells were adoptively transferred into TS1PKO mice intravenously at a dose of one million cells. TS1P1KO mice with reconstitution of CD4 T cells showed improved heart histology and less fibrosis as well as improved cardiac function compared with TS1P1KO mice without CD4 T cell transfer. In conclusion, deficiency of CD4 T cells in T cell S1P1 knockout mice contributes to cardiac fibrosis and myocardial dysfunction.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.