Improved therapeutic efficacy of mammalian expressed-recombinant interferon gamma against ovarian cancer cells
| Autor: | Leigh Owens, Vincent Jung, Kirsten Heimann, Narges Mashkour, Carina Villacrés, Michael Butler, Ali Razaghi |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Programmed cell death Glycosylation Blotting Western Biology Interferon-gamma 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Polysaccharides Cell Line Tumor medicine Humans Interferon gamma FADD Ovarian Neoplasms Cell Biology Recombinant Interferon Gamma Cytostasis Molecular biology Recombinant Proteins HEK293 Cells Treatment Outcome 030104 developmental biology chemistry Cell culture Apoptosis 030220 oncology & carcinogenesis biology.protein Cancer research Female medicine.drug |
| Zdroj: | Experimental Cell Research. 359:20-29 |
| ISSN: | 0014-4827 |
| DOI: | 10.1016/j.yexcr.2017.08.014 |
| Popis: | Human interferon gamma (hIFNγ) affects tumour cells and modulates immune responses, showing promise as an anti-cancer biotherapeutic. This study investigated the effect of glycosylation and expression system of recombinant hIFNγ in ovarian carcinoma cell lines, PEO1 and SKOV3. The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. Hydrophilic Interaction Liquid Chromatography determined the structure of N-linked glycans present in HEK293-expressed hIFNγ (hIFNγ-HEK). PEO1 was more sensitive to hIFNγ than SKOV3, but responses were dose-dependent and expression platform/glycosylation status-independent, whereas SKOV3 responded to mammalian-expressed hIFNγ in a dose-independent manner, only. Complex-type oligosaccharides dominated the N-glycosylation pattern of hIFNγ-HEK with some terminal sialylation and core fucosylation. Cleaved PARP and cleaved caspase-3 were not detected in either cell line, but FADD was expressed in SKOV3 with levels increased following treatment. In conclusion, hIFNγ did not induce apoptosis in either cell line. Mammalian- expressed hIFNγ increased cell death in the drug-resistant SKOV3. The presence of FADD in SKOV3, which may inhibit apoptosis through activation of NF-κB, could serve as a novel therapeutic target. |
| Databáze: | OpenAIRE |
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