2. Academic Validation
  3. Improved therapeutic efficacy of mammalian expressed-recombinant interferon gamma against ovarian cancer cells

Improved therapeutic efficacy of mammalian expressed-recombinant interferon gamma against ovarian cancer cells

  • Exp Cell Res. 2017 Oct 1;359(1):20-29. doi: 10.1016/j.yexcr.2017.08.014.
Ali Razaghi 1,  Carina Villacrés 2,  Vincent Jung 2,  Narges Mashkour 1,  Michael Butler 2,  Leigh Owens 1,  Kirsten Heimann 3
Affiliations

Affiliations

  • 1 Centre for Biodiscovery and Molecular Development of Therapeutics, James Cook University, Townsville QLD 4811, Australia.
  • 2 Department of Microbiology, University of Manitoba, Winnipeg, MB, Canada R3T 2N2.
  • 3 Centre for Biodiscovery and Molecular Development of Therapeutics, James Cook University, Townsville QLD 4811, Australia. Electronic address: Kirsten.heimann@jcu.edu.au.
PMID: 28803068 DOI: 10.1016/j.yexcr.2017.08.014
Abstract

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.

Keywords

FADD; Glycosylation; Interferon gamma; Ovarian cancer; SKOV3; Therapeutic efficacy.

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