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We evaluated the cytotoxic effect of isoleucine-zipper tumor necrosis factor-related apoptosis inducing ligand (izTRAIL) against cell lines, B101592, Cha, and C090115, derived from canine mammary gland tumors. These cells were derived from three dogs diagnosed with mammary adenoma or carcinoma. All three cells were positive for vimentin, while B101592 and C090115 were positive for cytokeratin (CK) AE1/AE3 and CK CAM5.2. Treatment with izTRAIL decreased the viability of the three cell lines. The proportion of annexin V+/propidium iodide- cells increased in all three cell lines after treatment with izTRAIL. Additionally, cell cycle analysis revealed that izTRAIL treatment increased the number of cells in sub-G1 phase. Moreover, izTRAIL treatment activated caspase-8 and caspase-3 and enhanced the levels of cleaved poly (ADP-ribose) polymerase. The cytotoxic effect of izTRAIL was mitigated upon co-treatment with caspase-8 or caspase-3 inhibitor. These results indicated that izTRAIL induces apoptosis in cell lines derived from canine mammary tumor, which was also previously reported in canine hemangiosarcoma cell lines. This suggested that canine tumor cells have conserved TRAIL receptors. This study will provide the basis for further studies on TRAIL receptors and TRAIL-related molecules.

Trimer form of tumor necrosis factor-related apoptosis inducing ligand induces apoptosis in canine cell lines derived from mammary tumors