DOCSLIB.ORG

Microrna-15A-5P Down-Regulation Inhibits Cervical Cancer by Targeting TP53INP1 in Vitro

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Microrna-15A-5P Down-Regulation Inhibits Cervical Cancer by Targeting TP53INP1 in Vitro European Review for Medical and Pharmacological Sciences 2019; 23: 8219-8229 MicroRNA-15a-5p down-regulation inhibits cervical cancer by targeting TP53INP1 in vitro X.-Q. ZHAO1, H. TANG2, J. YANG2, X.-Y. GU2, S.-M. WANG2, Y. DING2 1Medical School of Nantong University, Nantong, China 2Department of Endoscopic Diagnostic and Treatment Center, Women’s Hospital of Nanjing Medical University, Nanjing, China Abstract. – OBJECTIVE: An increasing num- Key Words: ber of reports have shown that microRNAs (miR- Cervical cancer, Apoptosis, MicroRNA-15a-5p, NAs) play a vital role in the occurrence and de- TP53INP1. velopment of cancer by acting as tumor inhib- itors or oncogenes. The purpose of this re- search was to explore whether the expression level of microRNA-15a-5p (miR-15a-5p) was re- Introduction lated to TP53 regulated inhibitor of apoptosis 1 (TP53INP1) in cervical cancer, and to explore the role of miR-15a-5p in cervical cancer in vitro. Cervical cancer, one of the most common PATIENTS AND METHODS: Human cervi- malignancies in females1,2, has become the main cal cancer tissues and adjacent normal tis- health issue of women for high morbidity and sues were obtained from 30 cervical cancer mortality3, especially in young women4,5. Com- patients. Firstly, we carried out the quantita- bined treatments have been a standard therapeu- tive Real Time-PCR (qRT-PCR) assay to eval- tic method for cervical cancer patients, including uate the level of miR-15a-5p in cervical can- 6 cer tissues and cell lines. The TargetScan and chemotherapy and radiotherapy . Also, radio- the Dual-Luciferase Reporter Assay were used therapy alone has been used for cervical cancer 7 to confirm the relationship between TP53INP1 at the early stage of cervical cancer . However, and miR-15a-5p. Besides, the Cell Counting Kit- low radiosensitivity, hypoxia, cellular glutathi- 8 (CCK-8) and the flow cytometry analysis were one, or other elements may result in the re-oc- performed to detect the effect of miR-15a-5p on currence of cervical cancer8,9. Thus, the basic cell proliferation and apoptosis in cervical can- research on the pathogenesis of cervical cancer is cer cells. very necessary to lessen the recurrence rate and RESULTS: Our results showed that the expres- sion of miR-15a-5p was enhanced in cervical advance the clinical treatment of cervical cancer. cancer tissues and cells lines. The data from the Particularly, the biological functions and the reg- Dual-Luciferase Reporter Assay demonstrated ulatory effects of the relative genes in cervical that TP53INP1 was a direct target of miR-15a-5p. cancer need to be explored. Besides, the specif- We also found that TP53INP1 was down-regulat- ic targets or genes for cervical cancer remain ed in the cervical cancer tissues and cell lines largely unknown. In this research, we investi- compared with the adjacent normal tissues and gated the molecular mechanism correlated with normal cervical cells. Besides, the down-reg- the tumorigenesis and development of cervical ulation of miR-15a-5p depressed cervical can- cer cell proliferation and enhanced cell apop- cancer and seek novel cure targets for cervical tosis. Our results clearly suggested that the cancer. MicroRNAs (miRNAs), a group of small down-regulation of TP53INP1 successfully im- non-coding RNAs with 20-22 nucleotides, can paired the tumor-inhibition effects of miR-15a- be directly combined with the 3′-untranslated re- 5p inhibitor in cervical cancer cells. gion (3′-UTR) of target genes to regulate the ex- CONCLUSIONS: Our findings indicated that pression of the target genes10. In various cancers, miR-15a-5p functioned as a tumor-promoting miRNAs can act as a part of oncogenic or tumor gene in cervical cancer by directly targeting inhibitors to regulate the tumor formation and TP53INP1, indicating that miR-15a-5p might be 11 a potential treatment target for cervical cancer the expression of their target genes . As we all patients. know, many studies have revealed that miRNAs Corresponding Author: Ye Ding, MD; e-mail: [email protected] 8219 X.-Q. Zhao, H. Tang, J. Yang, X.-Y. Gu, S.-M. Wang, Y. Ding play important roles in various cancer types, Cell Culture including colorectal cancer12, breast cancer13,14, The human cervical cancer cell lines HeLa, cervical cancer15-18, hepatic carcinoma19, and so SiHa, and the human normal cervical cell line on. Moreover, miRNAs are involved in cellular H8 were provided by the American Type Culture proliferation, differentiation, apoptosis, and pro- Collection (ATCC, Manassas, VA, USA). The cell gression. Thus, focusing on the unknown miR- lines were cultivated in the Roswell Park Memo- NAs could be a new insight into the treatment rial Institute-1640 (RPMI-1640; Gibco, Carlsbad, of cancer. Among plentiful tumors inhibiting CA, USA), supplemented with 10% fetal bovine miRNAs, miR-15a-5p belongs to miR-15a fam- serum (FBS; Gibco, Carlsbad, CA, USA), 1% ily and was found to regulate many biological penicillin/ streptomycin, and were incubated at behaviors, for example, cell proliferation, apop- 37°C in a humidified chamber supplemented with tosis, migration, and invasion in various human 5% CO2. cancer cell lines20-23. TP53INP1, TP53 regulated inhibitor of apoptosis 1, was down-regulated in Cell Transfection various cancers, correlates with poor prognosis HeLa and SiHa cells were seeded in the 6-well and cancer relapse24,25. However, the functional plates at a density of 2.0×106 per well and trans- roles of miR-15a-5p and TP53INP1 in human fected with the TP53INP1-siRNA, control-siR- cervical cancer are still unknown. Therefore, the NA, inhibitor control, miR-15a-5p inhibitor, or aim of this study was to investigate the effect of miR-15a-5p inhibitor+TP53INP1-siRNA for 48 miR-15a-5p and TP53INP1 on the progression h by Lipofectamine 2000 (Invitrogen, Carlsbad, of cervical cancer and to explore the poten- CA, USA) according to the manufacturer’s man- tial mechanism. In this paper, we found that ual. After transfection, the cells were collected miR-15a-5p was aberrantly up-regulated both in for further experiments. The efficiency of cell cervical cancer cell lines and clinical cervical transfection was detected by the qRT-PCR and cancer tissues. Then, we conducted molecular Western blot assay. biology and bioinformatics methods to suggest that TP53INP1 was the direct target of miR-15a- CCK-8 Assay 5p in cervical cancer. Besides, we observed that The Cell Counting Kit-8 (Sigma-Aldrich, St. the down-regulation of miR-15a-5p suppressed Louis, MO, USA) was carried out to measure cell cell proliferation and induced cell apoptosis in proliferation following the manufacturer’s proto- cervical cancer cells by targeting TP53INP1. cols. In brief, the cells were seeded in 96-well In addition, miR-15a-5p down-regulation could plates and cultured for 24 h. Then, the cells were also affect the relative gene expression, such transfected with the inhibitor control, miR-15a-3p as TP53INP1, Bcl-2, Bax, and Bcl-2, while its inhibitor, or miR-15a-3p inhibitor+TP53INP1-siR- effects could be reversed by TP53INP1-siRNA. NA for 48 h and then, we added 10 µl CCK-8 solu- Therefore, our results demonstrated important tion to each well and incubated them for another roles of miR-15a-5p in the pathogenesis of cervi- hour at 37°C. The microplate reader (Eon, Bio- cal cancer and indicated its potential functions in Teck, Winooski, VT, USA) was used to measure cervical cancer therapy. the optical density (OD) at 450 nm. Dual-Luciferase Reporter Assay Patients and Methods The TargetScan Release 7.1 (www.targetscan. org/vert_71) was used to predict the relationship Clinical Specimens Collection between miR-15a-5p and TP53INP1. The results The human cervical cancer tissues and the suggested that TP53INP1 was the potential target adjacent normal tissues were obtained from 30 of miR-15a-5p. Then, TP53INP1 3’-UTR DNA cervical cancer patients from the Women’s Hos- segments containing the target sequence of miR- pital of Nanjing Medical University. These spec- 15a-5p were inserted into a pmirGLO vector imens were stored in liquid nitrogen or at -80°C (Promega, Madison, WI, USA) to form the re- for future use. The experiment was approved by porter vector TP53INP1-wild-type (TP53INP1- the Ethics Committee of Women’s Hospital of WT). Also, the mutant 3’-UTRs of TP53INP1 Nanjing Medical University. All patients were was inserted into a Luciferase reporter vector notified that their specimens would be used in (pmiR-REPORT, Ambion, Austin, TX, USA) to this research. form TP53INP1-mutated-type (TP53INP1-MUT) 8220 MicroRNA-15a-5p down-regulation inhibits cervical cancer by targeting TP53INP1 in vitro constructs. Then, the mimic control or miR-15a- Western Blot Assay 5p mimic and TP53INP1-WT or TP53INP1-MUT The cervical cancer cells were washed with were co-transfected into 293 T cells with Lipo- ice-cold PBS and then lysed with RIPA buffer fectamine-2000 (Invitrogen, CA, USA) for 48 (Beyotime Biotechnology, Shanghai, China), h. The Dual-Luciferase Reporter Assay System centrifuged at 12.000 rpm for 30 min at 4°C. (Promega, Madison, WI, USA) was performed The Total Protein Extraction Kit (Phygene Life to detect the Luciferase activity according to Sciences, Shanghai, China) was employed to the manufacturer’s instructions. The experiments extract the total protein. The protein concentra- were carried out at least in triplicate. tion was evaluated by a BCA protein kit (Beyo- time Biotechnology, Haimen, China). Then, 40 Quantitative Real Time-PCR μg of protein were loaded onto 10% SDS-PAGE (qRT-PCR) Assay and then transferred into polyvinylidene diflu- The total RNA was extracted from cervi- oride (PVDF) membrane. Then, the membrane cal cancer cell lines or tissues using TRIzol was blocked with 5% non-fat milk in PBST (Invitrogen, Carlsbad, CA, USA) following the at room temperature for 1 h and incubated manufacturer’s instruction.
Load more

Recommended publications
  • Mir-155 Targets TP53INP1 to Regulate Liver Cancer Stem Cell Acquisition and Self-Renewal ⇑ Fengchao Liu, Xin Kong, Lin Lv, Jian Gao
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector FEBS Letters 589 (2015) 500–506 journal homepage: www.FEBSLetters.org MiR-155 targets TP53INP1 to regulate liver cancer stem cell acquisition and self-renewal ⇑ Fengchao Liu, Xin Kong, Lin Lv, Jian Gao Department of Gastroenterology, Second Affiliated Hospital, Chongqing Medical University, Chongqing, China article info abstract Article history: In liver cancer, miR-155 up-regulation can regulate cancer-cell invasion. However, whether miR-155 Received 29 October 2014 expression is associated with liver cancer stem cells (CSCs) remains unknown. Here, we show that Revised 5 January 2015 miR-155 expression is up-regulated in tumor spheres. Knock-down of miR-155 resulted in suppres- Accepted 9 January 2015 sion of tumor sphere formation, through a decrease in the proportion of CD90+ and CD133+ CSCs and Available online 17 January 2015 in the expression of Oct4, whereas miR-155 overexpression had the opposite effect. TP53INP1 was Edited by Quan Chen determined to be involved in the CSCs-like properties that were regulated by miR-155. Thus, miR- 155 may play an important role in promoting the generation of stem cell-like cells and their self- renewal by targeting the gene TP53INP1. Keywords: Liver cancer stem cell Ó 2015 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved. MiR-155 Tumor Protein 53-Induced Nuclear Protein 1 Tumor sphere Liver cancer 1. Introduction plants [10]. Recently, increasing studies have revealed that many miRNAs play crucial roles in tumorigenesis [11,12], including CSCs Hepatocellular carcinoma (HCC) is the sixth most prevalent [13,14].
    [Show full text]
  • Mir-504 Promotes Tumour Growth and Metastasis in Human Osteosarcoma by Targeting TP53INP1
    ONCOLOGY REPORTS 38: 2993-3000, 2017 miR-504 promotes tumour growth and metastasis in human osteosarcoma by targeting TP53INP1 QINGCHUN CAI, SIXIANG ZENG, XING DAI, JUNLONG WU and WEI MA Department of Orthopaedics, First Affiliated Hospital of the Medical College, Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China Received March 18, 2017; Accepted September 4, 2017 DOI: 10.3892/or.2017.5983 Abstract. An increasing number of studies have demonstrated and the second peak is at 75-79 years of age (1,2). The primary that microRNAs participate in the development of osteosar- tumour originates in the distal femur, proximal tibia or humerus coma by acting as tumour suppressor or tumour-promoting and its rapid cell division may contribute to the oncogenesis of genes. We investigated the role of miR-504 in the growth osteosarcoma during childhood (3). Adjuvant chemotherapy and metastasis of osteosarcoma. The expression of miR-504 following surgery, has improved the long-term survival rates in clinical osteosarcoma samples was higher than that in the from <20 to 70% in localized osteosarcoma. However, this adjacent normal tissue and correlated with tumour size and combined therapy has shown limited progress in the long-term clinical stage. Tumour protein p53-inducible nuclear protein 1 survival rates of metastatic osteosarcoma cases, which account (TP53INP1) was downregulated in the clinical osteosarcoma for one-quarter of all patients at the time of initial diagnosis. The samples compared with the adjacent normal tissues and was higher incidence of osteosarcoma in children and adolescents, consistently correlated with the clinical stage. The results its tendency to metastasize and the poor prognosis in metastatic of dual-luciferase reporter assay and western blot analysis cases render osteosarcoma the leading cause of cancer-related demonstrated that the TP53INP1 gene is a direct target of deaths in childhood, although it accounts only for 5% of child- miR-504.
    [Show full text]
  • TP53INP1 Down-Regulation Activates a P73-Dependent DUSP10/ERK Signaling Pathway To
    Author Manuscript Published OnlineFirst on July 3, 2017; DOI: 10.1158/0008-5472.CAN-16-3456 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. TP53INP1 down-regulation activates a p73-dependent DUSP10/ERK signaling pathway to promote metastasis of hepatocellular carcinoma Kai-Yu Ng1, Lok-Hei Chan1, Stella Chai1, Man Tong1, Xin-Yuan Guan2,4, Nikki P Lee3, Yunfei Yuan5, Dan Xie5, Terence K Lee6, Nelson J Dusetti7, Alice Carrier7, Stephanie Ma1,4 1School of Biomedical Sciences, Departments of 2Clincial Oncology and 3Surgery, 4State Key Laboratory for Liver Research, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong; 5State Key Laboratory of Oncology in Southern China, Sun Yat-Sen University Cancer Center, Guangzhou, China; 6Department of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Hong Kong; 7Aix Marseille University, CNRS, INSERM, Institut Paoli- Calmettes, CRCM, Marseille, France Corresponding author: Stephanie Ma, PhD, School of Biomedical Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong, 1/F, Laboratory Block, 21 Sassoon Road, Pok Fu Lam, Hong Kong. E-mail: [email protected]; Tel: 852-3917-9238; Fax: 852-2817-0857 Short title: TP53INP1 in HCC metastasis Keywords: metastasis; TP53INP1; p73; DUSP10; ERK; liver cancer Abbreviations: DUSP, dual-specificity MAP kinase phosphatases; EV, empty vector; HCC, hepatocellular carcinoma; IHC, immunohistochemistry; KD, knockdown; MKP, MAP kinase phosphatases; NTC, non-target control; OE, overexpression; qRT-PCR, quantitative real time 1 Downloaded from cancerres.aacrjournals.org on October 1, 2021. © 2017 American Association for Cancer Research. Author Manuscript Published OnlineFirst on July 3, 2017; DOI: 10.1158/0008-5472.CAN-16-3456 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
    [Show full text]
  • Rna-Sequencing Applications: Gene Expression Quantification and Methylator Phenotype Identification
    The Texas Medical Center Library DigitalCommons@TMC The University of Texas MD Anderson Cancer Center UTHealth Graduate School of The University of Texas MD Anderson Cancer Biomedical Sciences Dissertations and Theses Center UTHealth Graduate School of (Open Access) Biomedical Sciences 8-2013 RNA-SEQUENCING APPLICATIONS: GENE EXPRESSION QUANTIFICATION AND METHYLATOR PHENOTYPE IDENTIFICATION Guoshuai Cai Follow this and additional works at: https://digitalcommons.library.tmc.edu/utgsbs_dissertations Part of the Bioinformatics Commons, Computational Biology Commons, and the Medicine and Health Sciences Commons Recommended Citation Cai, Guoshuai, "RNA-SEQUENCING APPLICATIONS: GENE EXPRESSION QUANTIFICATION AND METHYLATOR PHENOTYPE IDENTIFICATION" (2013). The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences Dissertations and Theses (Open Access). 386. https://digitalcommons.library.tmc.edu/utgsbs_dissertations/386 This Dissertation (PhD) is brought to you for free and open access by the The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences at DigitalCommons@TMC. It has been accepted for inclusion in The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences Dissertations and Theses (Open Access) by an authorized administrator of DigitalCommons@TMC. For more information, please contact [email protected]. RNA-SEQUENCING APPLICATIONS: GENE EXPRESSION QUANTIFICATION AND METHYLATOR PHENOTYPE IDENTIFICATION
    [Show full text]
  • Oxidative Stress-Induced P53 Activity Is Enhanced by a Redox-Sensitive TP53INP1 Sumoylation
    Cell Death and Differentiation (2014) 21, 1107–1118 & 2014 Macmillan Publishers Limited All rights reserved 1350-9047/14 www.nature.com/cdd Oxidative stress-induced p53 activity is enhanced by a redox-sensitive TP53INP1 SUMOylation S Peuget1,2, T Bonacci1,2, P Soubeyran1, J Iovanna1 and NJ Dusetti*,1 Tumor Protein p53-Induced Nuclear Protein 1 (TP53INP1) is a tumor suppressor that modulates the p53 response to stress. TP53INP1 is one of the key mediators of p53 antioxidant function by promoting the p53 transcriptional activity on its target genes. TP53INP1 expression is deregulated in many types of cancers including pancreatic ductal adenocarcinoma in which its decrease occurs early during the preneoplastic development. In this work, we report that redox-dependent induction of p53 transcriptional activity is enhanced by the oxidative stress-induced SUMOylation of TP53INP1 at lysine 113. This SUMOylation is mediated by PIAS3 and CBX4, two SUMO ligases especially related to the p53 activation upon DNA damage. Importantly, this modification is reversed by three SUMO1-specific proteases SENP1, 2 and 6. Moreover, TP53INP1 SUMOylation induces its binding to p53 in the nucleus under oxidative stress conditions. TP53INP1 mutation at lysine 113 prevents the pro-apoptotic, antiproliferative and antioxidant effects of TP53INP1 by impairing the p53 response on its target genes p21, Bax and PUMA. We conclude that TP53INP1 SUMOylation is essential for the regulation of p53 activity induced by oxidative stress. Cell Death and Differentiation (2014) 21, 1107–1118; doi:10.1038/cdd.2014.28; published online 7 March 2014 Maintaining homeostasis in response to the broad range of stress is strictly dependent on p53, and TP53INP1-deficient intrinsic and extrinsic aggressions is a challenge for cells.
    [Show full text]
  • TP53INP1, a Tumor Suppressor, Interacts with LC3 and ATG8-Family Proteins Through the LC3-Interacting Region (LIR) and Promotes Autophagy-Dependent Cell Death
    Cell Death and Differentiation (2012) 19, 1525–1535 & 2012 Macmillan Publishers Limited All rights reserved 1350-9047/12 www.nature.com/cdd TP53INP1, a tumor suppressor, interacts with LC3 and ATG8-family proteins through the LC3-interacting region (LIR) and promotes autophagy-dependent cell death M Seillier1,2,4, S Peuget1,2,4, O Gayet1,2, C Gauthier1,2, P N’Guessan1,2, M Monte3, A Carrier1,2, JL Iovanna1,2 and NJ Dusetti*,1,2 TP53INP1 (tumor protein 53-induced nuclear protein 1) is a tumor suppressor, whose expression is downregulated in cancers from different organs. It was described as a p53 target gene involved in cell death, cell-cycle arrest and cellular migration. In this work, we show that TP53INP1 is also able to interact with ATG8-family proteins and to induce autophagy-dependent cell death. In agreement with this finding, we observe that TP53INP1, which is mainly nuclear, relocalizes in autophagosomes during autophagy where it is eventually degraded. TP53INP1-LC3 interaction occurs via a functional LC3-interacting region (LIR). Inactivating mutations of this sequence abolish TP53INP1-LC3 interaction, relocalize TP53INP1 in autophagosomes and decrease TP53INP1 ability to trigger cell death. Interestingly, TP53INP1 binds to ATG8-family proteins with higher affinity than p62, suggesting that it could partially displace p62 from autophagosomes, modifying thereby their composition. Moreover, silencing the expression of autophagy related genes (ATG5 or Beclin-1) or inhibiting caspase activity significantly decreases cell death induced by TP53INP1. These data indicate that cell death observed after TP53INP1-LC3 interaction depends on both autophagy and caspase activity. We conclude that TP53INP1 could act as a tumor suppressor by inducing cell death by caspase- dependent autophagy.
    [Show full text]
  • Original Article Microrna-155 Acts As an Oncogene by Targeting the Tumor Protein 53-Induced Nuclear Protein 1 in Esophageal Squamous Cell Carcinoma
    Int J Clin Exp Pathol 2014;7(2):602-610 www.ijcep.com /ISSN:1936-2625/IJCEP1312038 Original Article microRNA-155 acts as an oncogene by targeting the tumor protein 53-induced nuclear protein 1 in esophageal squamous cell carcinoma Jie Zhang1*, Chen Cheng1*, Xiang Yuan2, Jiang-Tu He2, Qiu-Hui Pan2*, Fen-Yong Sun1* 1Department of Clinical Laboratory Medicine, Shanghai Tenth People’s Hospital of Tongji University, Shanghai, China, 200072; 2Department of Central Laboratory, Shanghai Tenth People’s Hospital of Tongji University, Shang- hai, China, 200072. *Equal contributors. Received December 12, 2013; Accepted December 31, 2013; Epub January 15, 2014; Published February 1, 2014 Abstract: MicroRNA-155 (miR-155) is overexpressed in many human cancers; however, the function of miR-155 is largely unknown in esophageal squamous cell carcinoma (ESCC). In the present study, we found that miR-155 is dra- matically increased in ESCC tissues compared with the paired adjacent normal tissues, which suggested that miR- 155 acts as an oncogene in ESCC. We predicted that tumor protein p53-induced nuclear protein 1 (TP53INP1) is a candidate target gene of miR-155 given that miR-155 expression decreased mRNA and protein levels of TP53INP1 as determined by RT-PCR and Western blot analysis. In addition, miR-155 and TP53INP1 showed a negative relation in ESCC tissues. Dual luciferase-based reporter assay indicated direct regulation of TP53INP1 by miR-155. Further- more, we demonstrated that RNA interference of TP53INP1 increased the proliferation and colonies formation of EC-1 cells. Up-regulation of TP53INP1 abrogated miR-155 induced growth in EC-1 cells and mutation of TP53INP1 in 3’-UTR restored the effects when co-transfected with miR-155.
    [Show full text]
  • Tp53inp1 Gene Is Implicated in Early Radiation Response in Human Fibroblast Cells
    Int. J. Mol. Sci. 2015, 16, 25450-25465; doi:10.3390/ijms161025450 OPEN ACCESS International Journal of Molecular Sciences ISSN 1422-0067 www.mdpi.com/journal/ijms Article TP53inp1 Gene Is Implicated in Early Radiation Response in Human Fibroblast Cells Nikolett Sándor 1,2, Boglárka Schilling-Tóth 1, Enikő Kis 1, Lili Fodor 3, Fruzsina Mucsányi 3, Géza Sáfrány 1 and Hargita Hegyesi 1,4,* 1 Division of Molecular Radiobiology, National Public Health Center—National Research Directorate for Radiobiology and Radiohygiene, Anna 5, Budapest 1221, Hungary; E-Mails: [email protected] (N.S.); [email protected] (B.S.-T.); [email protected] (E.K.); [email protected] (G.S.) 2 Doctoral School of Pathological Sciences, Semmelweis University, Üllői 26, Budapest 1089, Hungary 3 Department of Genetics, Cell and Immunobiology, Semmelweis University, Nagyvárad tér 4, Budapest 1089, Hungary; E-Mails: [email protected] (L.F.); [email protected] (F.M.) 4 Department of Morphology and Physiology, College of Health Care, Semmelweis University, Vas 17, Budapest 1089, Hungary * Author to whom correspondence should be addressed; E-Mail: [email protected]; Tel.: +36-1-482-2000 (ext. 150); Fax: +36-1-482-2010. Academic Editor: Terrence Piva Received: 30 June 2015 / Accepted: 20 October 2015 / Published: 23 October 2015 Abstract: Tumor protein 53-induced nuclear protein-1 (TP53inp1) is expressed by activation via p53 and p73. The purpose of our study was to investigate the role of TP53inp1 in response of fibroblasts to ionizing radiation. γ-Ray radiation dose-dependently induces the expression of TP53inp1 in human immortalized fibroblast (F11hT) cells.
    [Show full text]
  • Chemico-Biological Interactions 196 (2012) 89–95
    Chemico-Biological Interactions 196 (2012) 89–95 Contents lists available at ScienceDirect Chemico-Biological Interactions journal homepage: www.elsevier.com/locate/chembioint Exposure to sodium tungstate and Respiratory Syncytial Virus results in hematological/immunological disease in C57BL/6J mice ⇑ Cynthia D. Fastje a, , Kevin Harper a, Chad Terry a, Paul R. Sheppard b, Mark L. Witten a,1 a Steele Children’s Research Center, PO Box 245073, University of Arizona, Tucson, AZ 85724-5073, USA b Laboratory of Tree-Ring Research, PO Box 210058, University of Arizona, Tucson, AZ 85721-0058, USA article info abstract Article history: The etiology of childhood leukemia is not known. Strong evidence indicates that precursor B-cell Acute Available online 1 May 2011 Lymphoblastic Leukemia (Pre-B ALL) is a genetic disease originating in utero. Environmental exposures in two concurrent, childhood leukemia clusters have been profiled and compared with geographically Keywords: similar control communities. The unique exposures, shared in common by the leukemia clusters, have Tungsten been modeled in C57BL/6 mice utilizing prenatal exposures. This previous investigation has suggested Respiratory Syncytial Virus in utero exposure to sodium tungstate (Na2WO4) may result in hematological/immunological disease Childhood leukemia through genes associated with viral defense. The working hypothesis is (1) in addition to spontaneously and/or chemically generated genetic lesions forming pre-leukemic clones, in utero exposure to Na2WO4 increases genetic susceptibility to viral influence(s); (2) postnatal exposure to a virus possessing the 1FXXKXFXXA/V9 peptide motif will cause an unnatural immune response encouraging proliferation in the B-cell precursor compartment. This study reports the results of exposing C57BL/6J mice to Na2WO4 3 in utero via water (15 ppm, ad libetum) and inhalation (mean concentration PM5 3.33 mg/m ) and to Respiratory Syncytial Virus (RSV) within 2 weeks of weaning.
    [Show full text]
  • Open Moore Sarah P53network.Pdf
    THE PENNSYLVANIA STATE UNIVERSITY SCHREYER HONORS COLLEGE DEPARTMENT OF BIOCHEMISTRY AND MOLECULAR BIOLOGY A SYSTEMATIC METHOD FOR ANALYZING STIMULUS-DEPENDENT ACTIVATION OF THE p53 TRANSCRIPTION NETWORK SARAH L. MOORE SPRING 2013 A thesis submitted in partial fulfillment of the requirements for a baccalaureate degree in Biochemistry and Molecular Biology with honors in Biochemistry and Molecular Biology Reviewed and approved* by the following: Dr. Yanming Wang Associate Professor of Biochemistry and Molecular Biology Thesis Supervisor Dr. Ming Tien Professor of Biochemistry and Molecular Biology Honors Advisor Dr. Scott Selleck Professor and Head, Department of Biochemistry and Molecular Biology * Signatures are on file in the Schreyer Honors College. i ABSTRACT The p53 protein responds to cellular stress, like DNA damage and nutrient depravation, by activating cell-cycle arrest, initiating apoptosis, or triggering autophagy (i.e., self eating). p53 also regulates a range of physiological functions, such as immune and inflammatory responses, metabolism, and cell motility. These diverse roles create the need for developing systematic methods to analyze which p53 pathways will be triggered or inhibited under certain conditions. To determine the expression patterns of p53 modifiers and target genes in response to various stresses, an extensive literature review was conducted to compile a quantitative reverse transcription polymerase chain reaction (qRT-PCR) primer library consisting of 350 genes involved in apoptosis, immune and inflammatory responses, metabolism, cell cycle control, autophagy, motility, DNA repair, and differentiation as part of the p53 network. Using this library, qRT-PCR was performed in cells with inducible p53 over-expression, DNA-damage, cancer drug treatment, serum starvation, and serum stimulation.
    [Show full text]
  • TP53INP1 (H-110): Sc-68919
    SAN TA C RUZ BI OTEC HNOL OG Y, INC . TP53INP1 (H-110): sc-68919 BACKGROUND APPLICATIONS TP53INP1 (tumor protein p53-inducible nuclear protein 1), also known as TP53INP1 (H-110) is recommended for detection of TP53INP1 of human, p53DINP1, SIP or Teap, is a 240 amino acid protein that localizes to nu- mouse and, to a lesser extent, rat origin by Western Blotting (starting dilu - clear bodies and exists as 2 alternatively spliced isoforms, designated tion 1:200, dilution range 1:100-1:1000), immunoprecipitation [1-2 µg per p53DINP1a and p53DINP1b. Expressed ubiquitously with higher expression 100-500 µg of total protein (1 ml of cell lysate)], immunofluorescence (start ing in testis, pancreas and spleen tissue, TP53INP1 functions in response to dilution 1:50, dilution range 1:50-1:500), immunohistochemistry (includ ing double-stranded DNA breaks and regulates p53-mediated apoptosis, specifi - paraffin-embedded sections) (starting dilution 1:50, dilution range 1:50- 1:500) cally by phosphorylating human p53 at Ser 46, an event that leads to cell and solid phase ELISA (starting dilution 1:30, dilution range 1:30-1:3000). death. Additionally, TP53INP1 is thought to interact with p73 and may be TP53INP1 (H-110) is also recommended for detection of TP53INP1 in addi - involved in the regulation of p73-controlled cell cycle progression. TP53INP1 tional species, including equine, canine, bovine and porcine. expression is downregulated in pancreatic ductal adenocarcinomas, sug - gesting that, via its ability to induce cell death, TP53INP1 plays a role in Suitable for use as control antibody for TP53INP1 siRNA (h): sc-76715, tumor suppression.
    [Show full text]
  • Table S1. 103 Ferroptosis-Related Genes Retrieved from the Genecards
    Table S1. 103 ferroptosis-related genes retrieved from the GeneCards. Gene Symbol Description Category GPX4 Glutathione Peroxidase 4 Protein Coding AIFM2 Apoptosis Inducing Factor Mitochondria Associated 2 Protein Coding TP53 Tumor Protein P53 Protein Coding ACSL4 Acyl-CoA Synthetase Long Chain Family Member 4 Protein Coding SLC7A11 Solute Carrier Family 7 Member 11 Protein Coding VDAC2 Voltage Dependent Anion Channel 2 Protein Coding VDAC3 Voltage Dependent Anion Channel 3 Protein Coding ATG5 Autophagy Related 5 Protein Coding ATG7 Autophagy Related 7 Protein Coding NCOA4 Nuclear Receptor Coactivator 4 Protein Coding HMOX1 Heme Oxygenase 1 Protein Coding SLC3A2 Solute Carrier Family 3 Member 2 Protein Coding ALOX15 Arachidonate 15-Lipoxygenase Protein Coding BECN1 Beclin 1 Protein Coding PRKAA1 Protein Kinase AMP-Activated Catalytic Subunit Alpha 1 Protein Coding SAT1 Spermidine/Spermine N1-Acetyltransferase 1 Protein Coding NF2 Neurofibromin 2 Protein Coding YAP1 Yes1 Associated Transcriptional Regulator Protein Coding FTH1 Ferritin Heavy Chain 1 Protein Coding TF Transferrin Protein Coding TFRC Transferrin Receptor Protein Coding FTL Ferritin Light Chain Protein Coding CYBB Cytochrome B-245 Beta Chain Protein Coding GSS Glutathione Synthetase Protein Coding CP Ceruloplasmin Protein Coding PRNP Prion Protein Protein Coding SLC11A2 Solute Carrier Family 11 Member 2 Protein Coding SLC40A1 Solute Carrier Family 40 Member 1 Protein Coding STEAP3 STEAP3 Metalloreductase Protein Coding ACSL1 Acyl-CoA Synthetase Long Chain Family Member 1 Protein
    [Show full text]