Susceptibility of the C2 canine mastocytoma cell line to the effects of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)

Abstract

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a member of the TNF family, which preferentially induces apoptosis in cells that have undergone malignant transformation. In humans, non-neoplastic cells are normally protected from the effects of TRAIL by expressing decoy receptors, lacking death domains. In contrast, neoplastic cells tend to downregulate their decoy receptor expression, increasing their susceptibility to the pro-apoptotic effects of TRAIL, via the functional TRAIL receptors. The aim of the current study was to investigate the effect of TRAIL on the canine C2 mastocytoma cell line to determine whether this agent might be a suitable treatment for mast cell tumors in dogs. C2 and MDCK cells were cultured with recombinant human TRAIL. Apoptosis was assessed using a Caspase 3 & 7 chemiluminescence assay and flow cytometry following Annexin V:FITC labelling. Cell metabolism was assessed using a colorimetric MTT-based assay. C2 cells demonstrated greater sensitivity to TRAILinduced apoptosis compared to MDCK cells by all assessment methods. The dog genome assembly was searched for orthologs of TRAIL and its receptors using published sequences from other species for reference. Although a canine ortholog for TRAIL was identified, only one TRAIL receptor ortholog (TNFRSF11B) could be found. C2, but not MDCK, cells expressed mRNA for TNFRSF11B, detected by RT-PCR. In other species, TNFRSF11B is a decoy receptor, as even though it has a death domain it is secreted due to its lack of a transmembrane domain. The effect of TRAIL on the C2 cell line suggests that this cytokine might be suitable for treatment of mast cell tumors in dogs.

3,4-Dicaffeoylquinic Acid, a Major Constituent of Brazilian Propolis, Increases TRAIL Expression and Extends the Lifetimes of Mice Infected with the Influenza A Virus

“…However, the infected animals were protected even at early stages of the infectious period without increased TRAIL expression because 3,4-diCQA increases the viability of infected cells, as suggested by previous ex vivo experiments [31]. It must be noted that MDCK cells do not express the TRAIL receptor [44]. 3,4-diCQA induced TRAIL mRNA at 7 dpi relative to the control group (Figure 3).…”

Tumour necrosis factor‐related apoptosis‐inducing ligand induces apoptosis in canine hemangiosarcoma cells in vitro

“…The human and dog TRAIL sequence identity is 81.3%, and the amino acid sequences of the trimeric or receptor‐binding domains of TRAIL are highly conserve . A previous study showed that rhTRAIL, the same as TRAIL‐TEC used in this study, induced apoptosis in a canine mastocytoma cell line . Of the three rhTRAIL proteins examined, izTRAIL showed the strongest ability to induce apoptosis in canine HSA cells.…”

“…Therefore, many normal cells are resistant to apoptosis induced by TRAIL. In veterinary medicine, a study on the effect of TRAIL on canine mast cell tumour cell lines revealed a role of the activation of caspase‐3 . In addition, caspase‐8 was shown to be activated by flavopiridol (FVP) in lymphoma cell lines exhibiting TRAIL resistance, but it is not clear whether TRAIL activated caspase‐8 as well .…”

“…In addition, caspase‐8 was shown to be activated by flavopiridol (FVP) in lymphoma cell lines exhibiting TRAIL resistance, but it is not clear whether TRAIL activated caspase‐8 as well . At present, the only TRAIL receptor identified in dogs is OPG, and its transfer signals have only recently been predicted based on gene sequence analysis . Therefore, how TRAIL receptors act in canine tumour cell lines and the mechanism of action of TRAIL remain unclear.…”

“…18,23 Therefore, various strategies for activation of the TRAIL pathway can be selected. In veterinary medicine, the effect of TRAIL has been evaluated in canine mast cell tumours, canine osteosarcoma cell lines, lymphoma cell lines and mammary cell lines; [24][25][26][27] however, the mechanism of action of TRAIL in canine tumour cells has not been elucidated. Moreover, TRAIL resistance can be acquired in several types of human tumour cells, and the underlying mechanism has been clarified to some extent using various types of cell lines, whereas less is known about this mechanism in canine cancers.…”