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Inhibition of CYP17 expression by adrenal androgens and transforming growth factor β in adrenocortical cells.

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Biernacka-Łukanty J. M., Lehmann T. P., Trzeciak W. H.
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vol. 51
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issue 4
907-917
EN
Cytochrome P450c17, encoded by the CYP17 gene, is a component of the 17a-hydroxylase/17,20-lyase enzyme complex essential for production of adrenal glucocorticoids and androgens as well as gonadal androgens. The expression of CYP17 in adrenocortical cells is stimulated by corticotropin (ACTH) via the signal transduction pathway involving cAMP and protein kinase A (PKA). Thus, in addition to glucocorticoids, ACTH stimulates formation of adrenal androgens, which are known to induce transforming growth factor β (TGF-β) secretion. TGF-β in turn inhibits steroid hormone output by attenuating both basal and ACTH-dependent expression of CYP17. The present study revealed that treatment of bovine and human H295R adrenocortical cells with androgens resulted in a decrease in the basal level of CYP17 transcript and cortisol secretion, without affecting forskolin-stimulated levels. We also demonstrated that in H295R cells TGF-β inhibited both basal and forskolin-stimulated accumulation of CYP17 mRNA. Determination of promoter activity, directing luciferase reporter gene expression in H295R cells transfected with deletion fragments of bovine CYP17 promoter, indicated that the -483 to -433 bp fragment of the promoter was necessary for the inhibitory action of TGF-β on CYP17 expression. It is concluded that in bovine and human adrenocortical cells, androgens inhibit basal CYP17 expression probably at the transcriptional level and independently of the effect of TGF-β.
2
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Expression of soluble recombinant TGF-β type II receptor fused with the Fc portion of human IgG1 (sTβRII-Fc) in NS0 cells

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Kwiatkowska E. P., Kazimierczak U., Mackiewicz A., Kowalczyk D. W.
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2006
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vol. 53
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issue 2
361-369
EN
We have constructed and expressed recombinant chimeric soluble TGF-β type II receptor fused with the Fc portion of human IgG1 (sTβRII-Fc) in NS0 mouse myeloma cells and isolated cell lines constitutively secreting very high levels of biologically active protein. The GS-NS0 expression system takes advantage of the strong human cytomegalovirus immediate early promoter expression vector and glutamine synthetase as a selectable marker. The recombinant chimeric receptor could be produced in high amounts and efficiently purified by one step chromatography on a protein A column. Biochemical studies revealed that recombinant sTβRII-Fc binds native TGF-β1 and TGF-β3 isoforms and neutralizes their activity in vitro.
3
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Immediate and Delayed Surgical Repair of Duodenal Defects in Rats with Small Intestinal Submucosa Patch

86%
Miliaras D., Miliaras S., Meditskou S., Papamitsou T.
Polish Journal of Surgery
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2015
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vol. 86
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issue 3
116-121
EN
Duodenal injuries, though rare, carry high rates of morbidity and mortality. The aim of the study was to evaluate the healing of the duodenal wall with the use of a Small Intestinal Submucosa (SIS) patch. Material and methods. We studied 40 Whistar-Albino rats divided into two groups. In group A, we created a small defect in the duodenal wall, which was immediately covered with a SIS patch. In group B, the SIS patch was sutured over the defect after 6-8 hours, in order to induce peritonitis. The animals of both groups were sacrificed after 2, 6, 12 and 16 weeks respectively. In addition, we studied the immunohistochemical expression of TGF-β, which is a major constituent of SIS, and plays a central role in the healing process. Results. Histology showed progressive development of the layers of the duodenal wall over the patch as early as the 2nd week in some of the animals of group A. Mucosa developed later on in the animals of group B, presumably due to the more intense inflammation elicited by peritonitis. Expression of TGF-β was initially more pronounced in the epithelial cells of the regenerating mucosa of animals of group A, but it was maintained in higher levels in the animals of group B, which showed delayed mucosa degeneration. Conclusions. SIS appears to be both efficient and safe for the management of duodenal trauma. TGF-β seems to play an important role in the healing process, inducing regeneration of the stroma, and controling epithelial growth.
4
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Angiogenic cytokines VEGF, TGF-β1, IL-8 and TNF secretion by human ovarian cancer cells

86%
Sienko J., Grabowska-Derlatka L., Czajkowski K.
Current Gynecologic Oncology
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2017
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vol. 15
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issue 2
99-104
EN
Objectives: Angiogenesis is a process that is indispensable in cancer progression. A complex network of tumor and microenvironment stimuli regulate angiogenesis. VEGF, TGF-β1, IL-8 and TNF belong to the angiogenic factors that are key points in vessel formation. The aim of the study was to assess h-VEGF, TGF-β1, IL-8 and TNF secretion by human ovarian cell lines. Material and methods: OVA 2, OVA 4, OVA 9, OVA 11 and OVA 14 cell lines were established in our laboratory. The cells derived from primary and metastatic tumors of epithelial and non-epithelial origin. SK-OV-3, MDAH 2774, CAOV-1 and OVP-10 were the cell lines obtained from other sources. The concentration of VEGF, TGF-β1 and IL-8 was determined in culture supernatants by using the ELISA tests. Results: OVA 11 secreted all the evaluated cytokines. MDAH 2774 was the source of h-VEGF, TGF-β1, IL-8. SK-OV-3 secreted h-VEGF and IL-8. OVA 4 secreted TGF-β1 and TNF. TNF was the only studied cytokine secreted by CAOV-1, OVA 2 and OVA 9 cell lines. OVA 14 did not secret any of the cytokines. Conclusions: The investigated cell lines present heterogeneous profile of angiogenic cytokine secretion and seem to be an interesting set of models for the study of angiogenic signaling, or target therapy.
PL
Cel: Angiogeneza jest procesem niezbędnym do progresji raka. Złożona sieć bodźców pochodzących od guza i z mikrośrodowiska reguluje angiogenezę. VEGF, TGF-β1, IL-8 i TNF należą do czynników angiogennych, które odgrywają kluczową rolę w tworzeniu naczyń. Celem pracy była ocena wydzielania h-VEGF, TGF-β1, IL-8 i TNF przez ludzkie linie raka jajnika. Materiał i metoda: Linie OVA 2, OVA 4, OVA 9, OVA 11 oraz OVA 14 zostały ustalone samodzielnie. Komórki pochodziły z pierwotnych lub przerzutowych guzów jajnika pochodzenia nabłonkowego lub nienabłonkowego. Linie SK-OV-3, MDAH 2774, CAOV-1 oraz OVP-10 pochodziły z innych źródeł. Stężenie VEGF, TGF-β1 i IL-8 określano w supernatantach hodowli komórkowych w teście ELISA. Wyniki: Linia OVA 11 wydzielała wszystkie badane cytokiny. Linia MDAH 2774 była źródłem h-VEGF, TGF-β1, IL-8. Linia SK-OV-3 wydzielała h-VEGF oraz IL-8. Linia OVA 4 wydzielała TGF-β1 i TNF. TNF był jedyną cytokiną wydzielaną przez linie CAOV-1, OVA 2 oraz OVA 9. Linia OVA 14 nie wydzielała żadnej spośród badanych cytokin. Wnioski: Badane linie komórkowe stanowią heterogenną grupę nowotworów wydzielających cytokiny o właściwościach angiogennych i wydają się interesującym panelem do badań nad procesami angiogenezy czy terapii celowanej.
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