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

Abstract

Tumour necrosis factor‐related apoptosis‐inducing ligand (TRAIL) is an apoptosis‐inducing cytokine that shows potential therapeutic value for human neoplasms, and is effective in some canine tumours; however, its potential for killing canine hemangiosarcoma (HSA) cells is unknown. Thus, we evaluated the proapoptotic effect of TRAIL in nine canine HSA cell lines. Cells (JuA1, JuB2, JuB2‐1, JuB4, Re11, Re12, Re21, Ud2 and Ud6) were cultured with three recombinant human TRAILs (rhTRAILs): TRAIL‐TEC derived from Escherichia coli, TRAIL‐TL derived from mammalian cells and isoleucine zipper recombinant human TRAIL (izTRAIL) containing an isoleucine‐zippered structure that facilitates trimerization. TRAIL‐TEC did not decrease the cell viability in any of the cell lines tested, whereas the other two rhTRAILs effectively decreased the viability of all cell lines as assessed by the WST‐1 assay. In canine HSA cells, izTRAIL induced apoptosis more effectively than TRAIL‐TL. In JuB4, Re12, and Ud6 cells, izTRAIL increased the activation of caspase‐3 and caspase‐8 and caused poly (ADP‐ribose) polymerase degradation. Moreover, izTRAIL treatment increased the proportion of Annexin V+/ Propidium iodide (PI)− apoptotic cells and nuclear fragmentation in izTRAIL‐sensitive cells. These results show that rhTRAIL can induce apoptosis in canine HSA cells, but the sensitivity of TRAIL was different depending on the cell lines. Therefore, TRAIL could be an effective therapeutic agent against canine HSA, but the specific mechanism of resistance should be determined to clarify under what conditions this treatment would be most effective.

Tumour necrosis factor‐related apoptosis‐inducing ligand (TRAIL) loss in canine mammary carcinoma

“…Consistent downregulation of both TRAIL mRNA and protein levels, and the positive correlation between TRAIL and downstream molecules, FADD and caspase-3, may imply that TRAILinduced extrinsic apoptotic signalling is disrupted in canine mammary carcinoma, even though the exact contribution of this pathway to ultimate tumorigenesis is questionable. However, it might be speculated that endogenous TRAIL suppresses the tumour development or progression in canine tumours while sparing normal cells, from the evidence of direct apoptosis-inducing effect of TRAIL to various cancer cell lines including mammary tumour, osteosarcoma, lymphoma and hemangiosarcoma in previous studies 21,22,32,33 and from the loss of In addition, as a relationship between TRAIL and its downstream molecules was observed, it could be suggested that the canine extrinsic apoptotic pathway may be organized in a similar manner to that in humans, even though the exact cellular mechanism, including TRAIL receptors, is unknown.…”

New molecular targets in canine hemangiosarcoma—Comparative review and future of the precision medicine

“…231 TRAIL and its receptor belong to the p53-independent extrinsic pathway of apoptosis induction and can lead to apoptosis independently of p53, commonly inactivated in HSA. 231 Thus far, it has been established in HSA that caspase-3 and -8 are at least partially involved in TRAIL-induced apoptosis. 231 In terms of Bcl-2 overexpression, also reported in HSA, 232 the only direct activator of caspase-3 is caspase-8.…”

“…231 Thus far, it has been established in HSA that caspase-3 and -8 are at least partially involved in TRAIL-induced apoptosis. 231 In terms of Bcl-2 overexpression, also reported in HSA, 232 the only direct activator of caspase-3 is caspase-8. 233 The expression of antiapoptotic factors, such as Bcl-2 and survivin, may contribute to the TRAIL-resistance in HSA.…”

“…233 The expression of antiapoptotic factors, such as Bcl-2 and survivin, may contribute to the TRAIL-resistance in HSA. 231…”

The immunohistochemical detection of peroxiredoxin 1 and 2 in canine spontaneous vascular endothelial tumors

“…To validate cross-reactivity of anti-human PRDX1 or 2 antibodies used in the present study to canine PRDX1 or 2 proteins by western blotting, HeLa which expressed PRDX1 and 2 [9,16] and Re 12 which was derived from canine HSA [27] were selected. SDS-PAGE and western blotting was performed according to previous report [8]. Briefly, protein lysates were prepared, and electrophoresed with SDS-PAGE and then transferred onto polyvinylidene difluoride membranes (Cytiva, Tokyo, Japan).…”