#PAGE_PARAMS# #ADS_HEAD_SCRIPTS# #MICRODATA#
preLekára.sk / Odborné časopisy / PLOS Genetics / 2013 - 2

Inflammation-Mediated Genetic and Epigenetic Alterations Drive Cancer Development in the Neighboring Epithelium upon Stromal Abrogation of TGF-β Signaling

English version České info

Deletion of tumor suppressor genes in stromal fibroblasts induces epithelial cancer development, suggesting an important role of stroma in epithelial homoeostasis. However, the underlying mechanisms remain to be elucidated. Here we report that deletion of the gene encoding TGFβ receptor 2 (Tgfbr2) in the stromal fibroblasts (Tgfbr2fspKO) induces inflammation and significant DNA damage in the neighboring epithelia of the forestomach. This results in loss or down-regulation of cyclin-dependent kinase inhibitors p15, p16, and p21, which contribute to the development of invasive squamous cell carcinoma (SCC). Anti-inflammation treatment restored p21 expression, delayed tumorigenesis, and increased survival of Tgfbr2fspKO mice. Our data demonstrate for the first time that inflammation is a critical player in the epigenetic silencing of p21 in tumor progression. Examination of human esophageal SCC showed a down-regulation of TGFβ receptor 2 (TβRII) in the stromal fibroblasts, as well as increased inflammation, DNA damage, and loss or decreased p15/p16 expression. Our study suggests anti-inflammation may be a new therapeutic option in treating human SCCs with down-regulation of TβRII in the stroma.

Vyšlo v časopise: Inflammation-Mediated Genetic and Epigenetic Alterations Drive Cancer Development in the Neighboring Epithelium upon Stromal Abrogation of TGF-β Signaling. PLoS Genet 9(2): e32767. doi:10.1371/journal.pgen.1003251
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1003251

Souhrn

Deletion of tumor suppressor genes in stromal fibroblasts induces epithelial cancer development, suggesting an important role of stroma in epithelial homoeostasis. However, the underlying mechanisms remain to be elucidated. Here we report that deletion of the gene encoding TGFβ receptor 2 (Tgfbr2) in the stromal fibroblasts (Tgfbr2fspKO) induces inflammation and significant DNA damage in the neighboring epithelia of the forestomach. This results in loss or down-regulation of cyclin-dependent kinase inhibitors p15, p16, and p21, which contribute to the development of invasive squamous cell carcinoma (SCC). Anti-inflammation treatment restored p21 expression, delayed tumorigenesis, and increased survival of Tgfbr2fspKO mice. Our data demonstrate for the first time that inflammation is a critical player in the epigenetic silencing of p21 in tumor progression. Examination of human esophageal SCC showed a down-regulation of TGFβ receptor 2 (TβRII) in the stromal fibroblasts, as well as increased inflammation, DNA damage, and loss or decreased p15/p16 expression. Our study suggests anti-inflammation may be a new therapeutic option in treating human SCCs with down-regulation of TβRII in the stroma.

Zdroje

1. BhowmickNA, ChytilA, PliethD, GorskaAE, DumontN, et al. (2004) TGF-beta signaling in fibroblasts modulates the oncogenic potential of adjacent epithelia. Science 303 : 848–851.

2. WallaceJA, LiF, LeoneG, OstrowskiMC (2011) Pten in the breast tumor microenvironment: modeling tumor-stroma coevolution. Cancer Res 71 : 1203–1207.

3. HillR, SongY, CardiffRD, Van DykeT (2005) Selective evolution of stromal mesenchyme with p53 loss in response to epithelial tumorigenesis. Cell 123 : 1001–1011.

4. TrimboliAJ, Cantemir-StoneCZ, LiF, WallaceJA, MerchantA, et al. (2009) Pten in stromal fibroblasts suppresses mammary epithelial tumours. Nature 461 : 1084–1091.

5. TanwarPS, ZhangL, RobertsDJ, TeixeiraJM (2011) Stromal deletion of the APC tumor suppressor in mice triggers development of endometrial cancer. Cancer Res 71 : 1584–1596.

6. DemehriS, TurkozA, KopanR (2009) Epidermal Notch1 loss promotes skin tumorigenesis by impacting the stromal microenvironment. Cancer Cell 16 : 55–66.

7. PatocsA, ZhangL, XuY, WeberF, CaldesT, et al. (2007) Breast-cancer stromal cells with TP53 mutations and nodal metastases. N Engl J Med 357 : 2543–2551.

8. BierieB, StoverDG, AbelTW, ChytilA, GorskaAE, et al. (2008) Transforming growth factor-beta regulates mammary carcinoma cell survival and interaction with the adjacent microenvironment. Cancer Res 68 : 1809–1819.

9. YangL (2010) TGFbeta and cancer metastasis: an inflammation link. Cancer Metastasis Rev 29 : 263–271.

10. BierieB, ChungCH, ParkerJS, StoverDG, ChengN, et al. (2009) Abrogation of TGF-beta signaling enhances chemokine production and correlates with prognosis in human breast cancer. J Clin Invest 119 : 1571–1582.

11. IjichiH, ChytilA, GorskaAE, AakreME, BierieB, et al. (2011) Inhibiting Cxcr2 disrupts tumor-stromal interactions and improves survival in a mouse model of pancreatic ductal adenocarcinoma. J Clin Invest 121 : 4106–4117.

12. KimBG, LiC, QiaoW, MamuraM, KasprzakB, et al. (2006) Smad4 signalling in T cells is required for suppression of gastrointestinal cancer. Nature 441 : 1015–1019.

13. FrancoOE, JiangM, StrandDW, PeacockJ, FernandezS, et al. (2011) Altered TGF-beta signaling in a subpopulation of human stromal cells promotes prostatic carcinogenesis. Cancer Res 71 : 1272–1281.

14. ChengN, ChytilA, ShyrY, JolyA, MosesHL (2008) Transforming growth factor-beta signaling-deficient fibroblasts enhance hepatocyte growth factor signaling in mammary carcinoma cells to promote scattering and invasion. Mol Cancer Res 6 : 1521–1533.

15. BlancoD, VicentS, FragaMF, Fernandez-GarciaI, FreireJ, et al. (2007) Molecular analysis of a multistep lung cancer model induced by chronic inflammation reveals epigenetic regulation of p16 and activation of the DNA damage response pathway. Neoplasia 9 : 840–852.

16. Mendez-AcunaL, Di TomasoMV, PalittiF, Martinez-LopezW (2010) Histone post-translational modifications in DNA damage response. Cytogenet Genome Res 128 : 28–36.

17. CrastaK, GanemNJ, DagherR, LantermannAB, IvanovaEV, et al. (2012) DNA breaks and chromosome pulverization from errors in mitosis. Nature 482 : 53–58.

18. LukasJ, LukasC, BartekJ (2011) More than just a focus: The chromatin response to DNA damage and its role in genome integrity maintenance. Nat Cell Biol 13 : 1161–1169.

19. KrimpenfortP, IjpenbergA, SongJY, van der ValkM, NawijnM, et al. (2007) p15Ink4b is a critical tumour suppressor in the absence of p16Ink4a. Nature 448 : 943–946.

20. BornsteinS, WhiteR, MalkoskiS, OkaM, HanG, et al. (2009) Smad4 loss in mice causes spontaneous head and neck cancer with increased genomic instability and inflammation. J Clin Invest 119 : 3408–3419.

21. StairsDB, BayneLJ, RhoadesB, VegaME, WaldronTJ, et al. (2011) Deletion of p120-catenin results in a tumor microenvironment with inflammation and cancer that establishes it as a tumor suppressor gene. Cancer Cell 19 : 470–483.

22. YangL, DebuskLM, FukudaK, FingletonB, Green-JarvisB, et al. (2004) Expansion of myeloid immune suppressor Gr+CD11b+ cells in tumor-bearing host directly promotes tumor angiogenesis. Cancer Cell 6 : 409–421.

23. GabrilovichDI, NagarajS (2009) Myeloid-derived suppressor cells as regulators of the immune system. Nat Rev Immunol 9 : 162–174.

24. PlottelCS, BlaserMJ (2011) Microbiome and malignancy. Cell Host Microbe 10 : 324–335.

25. YangL, FrancoisF, PeiZ (2012) Molecular pathways: pathogenesis and clinical implications of microbiome alteration in esophagitis and Barrett esophagus. Clin Cancer Res 18 : 2138–2144.

26. FukaiY, FukuchiM, MasudaN, OsawaH, KatoH, et al. (2003) Reduced expression of transforming growth factor-beta receptors is an unfavorable prognostic factor in human esophageal squamous cell carcinoma. Int J Cancer 104 : 161–166.

27. LiX, PlacencioV, IturreguiJM, UwamariyaC, Sharif-AfsharAR, et al. (2008) Prostate tumor progression is mediated by a paracrine TGF-beta/Wnt3a signaling axis. Oncogene 27 : 7118–7130.

28. LingY, HuangG, FanL, WeiL, ZhuJ, et al. (2011) CpG island methylator phenotype of cell-cycle regulators associated with TNM stage and poor prognosis in patients with oesophageal squamous cell carcinoma. J Clin Pathol 64 : 246–251.

29. BassAJ, WatanabeH, MermelCH, YuS, PernerS, et al. (2009) SOX2 is an amplified lineage-survival oncogene in lung and esophageal squamous cell carcinomas. Nat Genet 41 : 1238–1242.

30. XingEP, NieY, SongY, YangGY, CaiYC, et al. (1999) Mechanisms of inactivation of p14ARF, p15INK4b, and p16INK4a genes in human esophageal squamous cell carcinoma. Clin Cancer Res 5 : 2704–2713.

31. StrutzF, OkadaH, LoCW, DanoffT, CaroneRL, et al. (1995) Identification and characterization of a fibroblast marker: FSP1. J Cell Biol 130 : 393–405.

32. ScholtenD, OsterreicherCH, ScholtenA, IwaisakoK, GuG, et al. (2010) Genetic labeling does not detect epithelial-to-mesenchymal transition of cholangiocytes in liver fibrosis in mice. Gastroenterology 139 : 987–998.

33. OsterreicherCH, Penz-OsterreicherM, GrivennikovSI, GumaM, KoltsovaEK, et al. (2011) Fibroblast-specific protein 1 identifies an inflammatory subpopulation of macrophages in the liver. Proc Natl Acad Sci U S A 108 : 308–313.

34. BoomershineCS, ChamberlainA, KendallP, Afshar-SharifAR, HuangH, et al. (2009) Autoimmune pancreatitis results from loss of TGFbeta signalling in S100A4-positive dendritic cells. Gut 58 : 1267–1274.

35. InoueT, PliethD, VenkovCD, XuC, NeilsonEG (2005) Antibodies against macrophages that overlap in specificity with fibroblasts. Kidney Int 67 : 2488–2493.

36. MonteleoneG, MannJ, MonteleoneI, VavassoriP, BremnerR, et al. (2004) A failure of transforming growth factor-beta1 negative regulation maintains sustained NF-kappaB activation in gut inflammation. J Biol Chem 279 : 3925–3932.

37. EngleSJ, OrmsbyI, PawlowskiS, BoivinGP, CroftJ, et al. (2002) Elimination of colon cancer in germ-free transforming growth factor beta 1-deficient mice. Cancer Res 62 : 6362–6366.

38. PangB, ZhouX, YuH, DongM, TaghizadehK, et al. (2007) Lipid peroxidation dominates the chemistry of DNA adduct formation in a mouse model of inflammation. Carcinogenesis 28 : 1807–1813.

39. Padilla-NashHM, HathcockK, McNeilNE, MackD, HoeppnerD, et al. (2012) Spontaneous transformation of murine epithelial cells requires the early acquisition of specific chromosomal aneuploidies and genomic imbalances. Genes Chromosomes Cancer 51 : 353–374.

40. MeiraLB, BugniJM, GreenSL, LeeCW, PangB, et al. (2008) DNA damage induced by chronic inflammation contributes to colon carcinogenesis in mice. J Clin Invest 118 : 2516–2525.

41. JanssenA, van der BurgM, SzuhaiK, KopsGJ, MedemaRH (2011) Chromosome segregation errors as a cause of DNA damage and structural chromosome aberrations. Science 333 : 1895–1898.

42. HerzogCR, WisemanRW, YouM (1994) Deletion mapping of a putative tumor suppressor gene on chromosome 4 in mouse lung tumors. Cancer Res 54 : 4007–4010.

43. MontagnaC, AndrechekER, Padilla-NashH, MullerWJ, RiedT (2002) Centrosome abnormalities, recurring deletions of chromosome 4, and genomic amplification of HER2/neu define mouse mammary gland adenocarcinomas induced by mutant HER2/neu. Oncogene 21 : 890–898.

44. LiuY, HanZP, ZhangSS, JingYY, BuXX, et al. (2011) Effects of inflammatory factors on mesenchymal stem cells and their role in the promotion of tumor angiogenesis in colon cancer. J Biol Chem 286 : 25007–25015.

45. Resta-LenertS, BarrettKE (2006) Probiotics and commensals reverse TNF-alpha -⁠ and IFN-gamma-induced dysfunction in human intestinal epithelial cells. Gastroenterology 130 : 731–746.

46. TengY, SunAN, PanXC, YangG, YangLL, et al. (2006) Synergistic function of Smad4 and PTEN in suppressing forestomach squamous cell carcinoma in the mouse. Cancer Res 66 : 6972–6981.

47. AchyutBR, YangL (2011) Transforming growth factor-beta in the gastrointestinal and hepatic tumor microenvironment. Gastroenterology 141 : 1167–1178.

48. SuH, HuN, YangHH, WangC, TakikitaM, et al. (2011) Global gene expression profiling and validation in esophageal squamous cell carcinoma and its association with clinical phenotypes. Clin Cancer Res 17 : 2955–2966.

49. HuN, CliffordRJ, YangHH, WangC, GoldsteinAM, et al. (2010) Genome wide analysis of DNA copy number neutral loss of heterozygosity (CNNLOH) and its relation to gene expression in esophageal squamous cell carcinoma. BMC Genomics 11 : 576.

50. HashimotoN, InayamaM, FujishimaM, ShiozakiH (2007) Clinicopathologic significance of expression of cyclooxygenase-2 in human esophageal squamous cell carcinoma. Hepatogastroenterology 54 : 758–760.

51. XingEP, NieY, WangLD, YangGY, YangCS (1999) Aberrant methylation of p16INK4a and deletion of p15INK4b are frequent events in human esophageal cancer in Linxian, China. Carcinogenesis 20 : 77–84.

52. HuN, WangC, SuH, LiWJ, Emmert-BuckMR, et al. (2004) High frequency of CDKN2A alterations in esophageal squamous cell carcinoma from a high-risk Chinese population. Genes Chromosomes Cancer 39 : 205–216.

53. GlickAaVWC (2011) Signaling Pathways in Squamous Cancer Springer.

54. FinakG, BertosN, PepinF, SadekovaS, SouleimanovaM, et al. (2008) Stromal gene expression predicts clinical outcome in breast cancer. Nat Med 14 : 518–527.

55. BacmanD, MerkelS, CronerR, PapadopoulosT, BruecklW, et al. (2007) TGF-beta receptor 2 downregulation in tumour-associated stroma worsens prognosis and high-grade tumours show more tumour-associated macrophages and lower TGF-beta1 expression in colon carcinoma: a retrospective study. BMC Cancer 7 : 156.

56. GartnerK, PfaffJ (1979) The forestomach in rats and mice, a food store without bacterial protein digestion. Zentralbl Veterinarmed A 26 : 530–541.

57. MatheEA, NguyenGH, BowmanED, ZhaoY, BudhuA, et al. (2009) MicroRNA expression in squamous cell carcinoma and adenocarcinoma of the esophagus: associations with survival. Clin Cancer Res 15 : 6192–6200.

Štítky
Genetika Reprodukčná medicína
Článek Complex Inheritance of Melanoma and Pigmentation of Coat and Skin in Grey Horses
Článek A Meta-Analysis of Thyroid-Related Traits Reveals Novel Loci and Gender-Specific Differences in the Regulation of Thyroid Function
Článek Genetic Landscape of Open Chromatin in Yeast
Článek Deleterious Alleles in the Human Genome Are on Average Younger Than Neutral Alleles of the Same Frequency
Článek Age-Dependent Transition from Cell-Level to Population-Level Control in Murine Intestinal Homeostasis Revealed by Coalescence Analysis
Článek Next-Generation Sequencing Identifies the Danforth's Short Tail Mouse Mutation as a Retrotransposon Insertion Affecting Expression
Článek ImmunoChip Study Implicates Antigen Presentation to T Cells in Narcolepsy
Článek Massive Mitochondrial Gene Transfer in a Parasitic Flowering Plant Clade
Článek Comment on “Genomic Hypomethylation in the Human Germline Associates with Selective Structural Mutability in the Human Genome”
Článek The Prefoldin Bud27 Mediates the Assembly of the Eukaryotic RNA Polymerases in an Rpb5-Dependent Manner
Článek Genetic Determinants of Trabecular and Cortical Volumetric Bone Mineral Densities and Bone Microstructure
Článek Encodes a Novel and -Genus-Specific Regulator of Photoperiodic Flowering in Rice
Článek Only One Isoform of CTP Synthase Forms the Cytoophidium
Článek Mechanisms Involved in the Functional Divergence of Duplicated GroEL Chaperonins in DK1622
Článek A Genome-Wide RNAi Screen in Identifies the Nicotinic Acetylcholine Receptor Subunit ACR-7 as an Antipsychotic Drug Target
Článek Autophagy Induction Is a Tor- and Tp53-Independent Cell Survival Response in a Zebrafish Model of Disrupted Ribosome Biogenesis
Článek Ancient DNA Reveals Prehistoric Gene-Flow from Siberia in the Complex Human Population History of North East Europe
Článek MicroRNA-3148 Modulates Allelic Expression of Toll-Like Receptor 7 Variant Associated with Systemic Lupus Erythematosus
Článek RNAi–Based Functional Profiling of Loci from Blood Lipid Genome-Wide Association Studies Identifies Genes with Cholesterol-Regulatory Function
Článek CELF Family RNA–Binding Protein UNC-75 Regulates Two Sets of Mutually Exclusive Exons of the Gene in Neuron-Specific Manners in
Článek Coordination of Chromatid Separation and Spindle Elongation by Antagonistic Activities of Mitotic and S-Phase CDKs
Článek The Ubiquitin Ligase Subunit Acts in Target Tissue to Restrict Tracheal Terminal Cell Branching and Hypoxic-Induced Gene Expression
Článek Mitotic Evolution of Shows a Stable Core Genome but Recombination in Antigen Families
Článek Tysnd1 Deficiency in Mice Interferes with the Peroxisomal Localization of PTS2 Enzymes, Causing Lipid Metabolic Abnormalities and Male Infertility
Článek A Regulatory Pathway, Ecdysone-Transcription Factor Relish-Cathepsin L, Is Involved in Insect Fat Body Dissociation
Článek PcG-Mediated Higher-Order Chromatin Structures Modulate Replication Programs at the BX-C
Článek MSH3 Polymorphisms and Protein Levels Affect CAG Repeat Instability in Huntington's Disease Mice
Článek JNK-Interacting Protein 3 Mediates the Retrograde Transport of Activated c-Jun N-Terminal Kinase and Lysosomes
Článek Discovery of a Splicing Regulator Required for Cell Cycle Progression
Článek Rearrangements of 2.5 Kilobases of Noncoding DNA from the Locus Define Predictive Rules of Genomic -Regulatory Logic
Článek Admixture Mapping in Lupus Identifies Multiple Functional Variants within IFIH1 Associated with Apoptosis, Inflammation, and Autoantibody Production
Článek Roles of the Developmental Regulator Homothorax in Limiting Longevity in
Článek miR-199a-5p Is Upregulated during Fibrogenic Response to Tissue Injury and Mediates TGFbeta-Induced Lung Fibroblast Activation by Targeting Caveolin-1
Článek A Kinome-Wide RNAi Screen in Glia Reveals That the RIO Kinases Mediate Cell Proliferation and Survival through TORC2-Akt Signaling in Glioblastoma
Článek Assembly of the Auditory Circuitry by a Genetic Network in the Mouse Brainstem
Článek SOX2 Co-Occupies Distal Enhancer Elements with Distinct POU Factors in ESCs and NPCs to Specify Cell State
Článek Retrotransposon Activates Ectopic Expression: A Short Tail
Článek Confounding by Repetitive Elements and CpG Islands Does Not Explain the Association between Hypomethylation and Genomic Instability
Článek Cell Reprogramming Requires Silencing of a Core Subset of Polycomb Targets
Článek Properties and Modeling of GWAS when Complex Disease Risk Is Due to Non-Complementing, Deleterious Mutations in Genes of Large Effect
Článek Essential Developmental, Genomic Stability, and Tumour Suppressor Functions of the Mouse Orthologue of
Článek Conditional Inactivation of the DNA Damage Response Gene in Mouse Testis Reveals Separable Roles for Components of the RAD9-RAD1-HUS1 Complex in Meiotic Chromosome Maintenance
Článek Genome-Wide Analysis Points to Roles for Extracellular Matrix Remodeling, the Visual Cycle, and Neuronal Development in Myopia
Článek Patterning of Leaf Vein Networks by Convergent Auxin Transport Pathways
Článek An Evolutionary Perspective on Epistasis and the Missing Heritability
Článek A Retrotransposon Insertion in the 5′ Regulatory Domain of Ptf1a Results in Ectopic Gene Expression and Multiple Congenital Defects in Danforth's Short Tail Mouse
Článek The Mub1/Ubr2 Ubiquitin Ligase Complex Regulates the Conserved Dsn1 Kinetochore Protein
Článek Mutations Can Cause Enamel-Renal Syndrome (ERS)
Článek Yemanuclein and HIRA Cooperate for Assembly of H3.3-Containing Nucleosomes in the Male Pronucleus
Článek Hepatocyte Growth Factor, a Determinant of Airspace Homeostasis in the Murine Lung
Článek ISWI and CHD Chromatin Remodelers Bind Promoters but Act in Gene Bodies
Článek COM-1 Promotes Homologous Recombination during Meiosis by Antagonizing Ku-Mediated Non-Homologous End Joining
Článek Control of Multicellular Development by the Physically Interacting Deneddylases DEN1/DenA and COP9 Signalosome
Článek Antagonism Versus Cooperativity with TALE Cofactors at the Base of the Functional Diversification of Hox Protein Function
Článek Dynamic Association of NUP98 with the Human Genome
Článek Ectopic Expression of Induces Spinal Defects, Urogenital Defects, and Anorectal Malformations in Mice
Článek Regulation of Contributes to the Lineage Potential of Neurogenin3+ Endocrine Precursor Cells in the Pancreas
Článek Gene-Based Testing of Interactions in Association Studies of Quantitative Traits
Článek The Amidation Step of Diphthamide Biosynthesis in Yeast Requires , a Gene Identified through Mining the - Interaction Network
Článek Plant-Symbiotic Fungi as Chemical Engineers: Multi-Genome Analysis of the Clavicipitaceae Reveals Dynamics of Alkaloid Loci
Článek Genome-Wide Diversity in the Levant Reveals Recent Structuring by Culture
Článek DNA Methylation Mediated Control of Gene Expression Is Critical for Development of Crown Gall Tumors
Článek Identification of the SlmA Active Site Responsible for Blocking Bacterial Cytokinetic Ring Assembly over the Chromosome
Článek Expression of a Novel P22 ORFan Gene Reveals the Phage Carrier State in Typhimurium
Článek Altered Cohesin Gene Dosage Affects Mammalian Meiotic Chromosome Structure and Behavior
Článek Quantitative Analysis of Histone Modifications: Formaldehyde Is a Source of Pathological N-Formyllysine That Is Refractory to Histone Deacetylases
Článek Duplicate Abalone Egg Coat Proteins Bind Sperm Lysin Similarly, but Evolve Oppositely, Consistent with Molecular Mimicry at Fertilization
Článek Lessons from on the Strengths and Weaknesses of Structured Association Mapping
Článek DNA–Methylome Analysis of Mouse Intestinal Adenoma Identifies a Tumour-Specific Signature That Is Partly Conserved in Human Colon Cancer
Článek Transposon Variants and Their Effects on Gene Expression in
Článek Polygenic Modeling with Bayesian Sparse Linear Mixed Models
Článek Single Transmembrane Peptide DinQ Modulates Membrane-Dependent Activities
Článek The JNK Signaling Pathway Activates Expression of Stress Response Genes by Derepressing the Fos/HDAC Repressor Complex
Článek The Interaction of CtIP and Nbs1 Connects CDK and ATM to Regulate HR–Mediated Double-Strand Break Repair
Článek Regulation of Metamorphosis by Xenobiotic Response Regulators
Článok vyšiel v časopise

PLOS Genetics

2013 Číslo 2
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Kurzy

Zvýšte si kvalifikáciu online z pohodlia domova

nový kurz
Autori: doc. MUDr. David Zemánek, Ph.D., MUDr. Anna Chaloupka, Ph.D.
Všetky kurzy
Prihlásenie
Zabudnuté heslo

Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.

Prihlásenie

Nemáte účet?  Registrujte sa

#ADS_BOTTOM_SCRIPTS#