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A complete compilation of matrix metalloproteinase expression in human malignant gliomas

World Journal of Clinical Oncology 2012年第5期3卷 67-79页

作者： Carsten Hagemann jelena anacker Ralf-Ingo Ernestus Giles H Vince Department of Neurosurgery Tumorbiology Laboratory University of Würzburg Josef-Schneider-Str. 11 D-97080 Würzburg Germany Department of Obstetrics and Gynaecology University of Würzburg Josef-Schneider-Str. 4 D-97080 Würzburg Germany

Glioblastomas are characterized by an aggressive local growth pattern, a marked degree of invasiveness and poor prognosis. Tumor invasiveness is facilitated by the increased activity of proteolytic enzymes which are involved in destruction of the extracellular matrix of the surrounding healthy brain tissue. Elevated levels of matrix metalloproteinases(MMPs) were found in glioblastoma(GBM) cell-lines, as well as in GBM biopsies as compared with low-grade astrocytoma(LGA) and normal brain samples, indicating a role in malignant progression. A careful review of the available literature revealed that both the expression and role of several of the 23 human MMP proteins is controversely discussed and for some there are no data available at all. We therefore screened a panel of 15 LGA and 15 GBM biopsy samples for those MMPs for which there is either no, very limited or even contradictory dataavailable. Hence, this is the first complete compilation of the expression pattern of all 23 human MMPs in astrocytic tumors. This study will support a better understanding of the specific expression patterns and interaction of proteolytic enzymes in malignant human glioma and may provide additional starting points for targeted patient therapy.

关键词： Astrocytic tumor Expression pattern Glioblastoma cell-lines Glioblastoma multiforme Matrix metalloproteinase

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Hypoxia and cytokines regulate carbonic anhydrase 9 expression in hepatocellular carcinoma cells in vitro

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World Journal of Clinical Oncology 2012年第6期3卷 82-91页

作者： Feray Kockar Hatice Yildrim Rahsan Ilikci Sagkan Carsten Hagemann Yasemin Soysal jelena anacker Ahmed Ayad Hamza Dirk Vordermark Michael Flentje Harun M Said Department of Biology Faculty of Art and Science Balikesir University Department of Immunology and Allergy Diseases Gulhane Military Medical Academy Tumorbiology Laboratory Department of Neurosurgery University of Würzburg Faculty of Medicine Department of Medical Genetics Afyon Kocatepe University Department of Obstetrics and Gynaecology University of Würzburg School of Medicine Southern Illinois University Department of Radiation Oncology University-Halle-Wittenberg Department of Radiation Oncology University of Wuerzburg

AIM: To study the expression of carbonic anhydrase(CA) 9 in human hepatocellular carcinoma(HCC) ***: We studied CA9 protein, CA9 m RNA and hypoxia-inducible factor-1 alpha(HIF-1α) protein levels in Hep3 B cells exposed in different parallel approaches. In one of these approaches, HCC cells were exposed to extreme in vitro hypoxia(24 h 0.1% O2) without or with interleukin(IL)-1, IL-6, tumor necrosis factoralpha(TNF-α) and transforming growth factor-beta(TGF-β) stimulation for the same hypoxic exposure time or exposed to normoxic oxygenation conditions without or with cytokine ***: The tumour cell line analysed showed a strong hypoxic CA9 m RNA expression pattern in response to prolonged severe hypoxia with cell-line specific patterns and a marked induction of CA9 protein in response to severe hypoxia. These results were paralleled by the results for HIF-1α protein under identical oxygenation conditions with a similar expression tendency to that displayed during the CA9 protein expression experimental series. Continuous stimulation with the cytokines, IL-1, IL-6, TNF-α and TGF-β, under normoxic conditions significantly increased the carbonic anhydrase 9 expression level at both the protein and m RNA level, almost doubling the CA9 m RNA and CA9 and HIF-1α protein expression levels found under hypoxia. The findings from these experiments indicated that hypoxia is a positive regulator of CA9 expression in HCC, and the four signal transduction pathways, IL-1, IL-6, TNF-α and TGF-β, positively influence CA9 expression under both normoxic and hypoxic ***: These findings may potentially be considered in the design of anti- cancer therapeutic approaches involving hypoxia-induced or cytokine stimulatory effects on expression. In addition, they provideevidence of the stimulatory role of the examined cytokine families resulting in an increase in CA9 expression under different oxygenation conditions in human cancer, especially HCC, and

关键词： Angiogenesis Carbonic anhydrase 9 Hypoxia Hypoxia-inducible factor-1 alpha Oxygen Radiotherapy Transforming growth factor-beta Tumour microenviroment

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Inhibition of N-Myc down regulated gene 1 in in vitro cultured human glioblastoma cells

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World Journal of Clinical Oncology 2012年第7期3卷 104-110页

作者： Harun M Said Buelent Polat Susanne Stein Mathias Guckenberger Carsten Hagemann Adrian Staab Astrid Katzer jelena anacker Michael Flentje Dirk Vordermark Department of Radiation Oncology University of Wuerzburg 97080 Würzburg Germany Department of Hematology and Oncology Johannes Gutenberg University Ⅲ Medical School 55101 Mainz Germany Department of Neurosurgery Tumorbiology Laboratory University of Wuerzburg 97080 Würzburg Germany Department of Radiation Oncology Paul Scherer Institute CH-5100 Villingen Switzerland Department of Gynaecology and Obstetrics University of Wuerzburg 97080 Würzburg Germany Department of Radiation Oncology University-Halle-Wittenberg 06110 Halle Germany

AIM: To study short ds RNA oligonucleotides(si RNA)as a potent tool for artificially modulating gene expression of N-Myc down regulated gene 1(NDRG1) gene induced under different physiological conditions(Normoxia and hypoxia) modulating NDRG1 transcription, m RNA stability and translation. METHODS: A cell line established from a patient with glioblastoma multiforme. Plasmid DNA for transfections was prepared with the Endofree Plasmid Maxi kit. From plates containing 5 × 107 cells, nuclear extracts were prepared according to previous protocols. The p SUPERNDRG1 vectors were designed, two sequences were selected from the human NDRG1 c DNA(5'-GCATTATTGGCATGGGAAC-3' and 5'-ATGCAGAGTAACGTGGAAG-3'. reverse transcription polymerase chain reaction was performed using primers designed using published information on β-actin and hypoxia-inducible factor(HIF)-1α m RNA sequences in Gen Bank. NDRG1 m RNA and protein level expression results under different conditions of hypoxia or reoxygenation were compared to aerobic control conditions using the Mann-Whitney U test. Reoxygenation values were also compared to the NDRG1 levels after 24 h of hypoxia(P < 0.05 was considered significant).RESULTS: si RNA- and iodoacetate(IAA)-mediated downregulation of NDRG1 m RNA and protein expression in vitro in human glioblastoma cell lines showed a nearly complete inhibition of NDRG1 expression when compared to the results obtained due to the inhibitory role of glycolysis inhibitor IAA. Hypoxia responsive elements bound by nuclear HIF-1 in human glioblastoma cells in vitro under different oxygenation conditions and the clearly enhanced binding of nuclear extracts from glioblastoma cell samples exposed to extreme hypoxic conditions confirmed the HIF-1 Western blotting results. CONCLUSION: NDRG1 represents an additional diagnostic marker for brain tumor detection, due to the role of hypoxia in regulating this gene, and it canrepresent a potential target for tumor treatment in human glioblastoma. T

关键词： N-Myc down regulated gene 1 Short dsRNA oligonucleotides Human cancer diseases Brain cancer Radiotherapy

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