Anticancer effects of phenoxazine derivatives combined with tumor necrosis factor-related apoptosis-inducing ligand on pancreatic cancer cell lines, KLM-1 and MIA-PaCa-2

Abstract

The aim of this study was to investigate the anticancer effects of the phenoxazine derivatives, 2-amino-4,4·-dihydro-4·,7-dimethyl-3H-phenoxazine-3-one (Phx-1), 3-amino-1,4·-dihydro-4·,8-dimethyl-2H-phenoxazine-2-one (Phx-2), and 2-aminophenoxazine-3-one (Phx-3) on human pancreatic cancer cell lines, KLM-1 and MIA-PaCa-2, in combination with tumor necrosis factor-related apoptosisinducing ligand (TRAIL), a member of the tumor necrosis factor superfamily of cytokines. Of these three phenoxazines, Phx-1 and Phx-3 inhibited proliferation of KLM-1 dosedependently, but Phx-2 did not. Phx-3 caused both apoptosis and necrosis in KLM-1 cells, as evidenced by the phosphatidylserine externalization and propidium iodide permeable cells detected by a flow cytometric method using annexin-V and propidium iodide. Down-regulation of Bcl-2 expression appeared to be involved in the Phx-3-induced cell death. TRAIL did not affect proliferation of KLM-1, and the inhibitory effects of Phx-1 and Phx-3 on the KLM-1 cell line were not augmented by the combination with TRAIL. On the other hand, proliferation of the MIA-PaCa-2 cell line was not affected by Phx-1, Phx-2 and Phx-3, although it was significantly inhibited by TRAIL in a dose-dependent manner. Inhibitory effects of TRAIL on MIA-PaCa-2 were synergistically augmented by the addition of Phx-1 and Phx-3, but not by Phx-2. These results suggest that both Phx-1 and Phx-3 exert anticancer effects against human pancreatic cancer cells, KLM-1 and MIA-PaCa-2, through distinct action modes. Phx-1 and Phx-3 may be effective for the treatment of pancreatic cancer.

Apoptosis Induction Preceded by Mitochondrial Depolarization in Multiple Myeloma Cell Line U266 by 2-Aminophenoxazine-3-one

“…U266 cells (1ϫ10 6 cells/10 ml) were treated with 0, 50 and 100 mM Phx-3 in 25 cm 2 cell culture flasks. After 24, 48 and 72 h of treatment, the cells were collected, once washed with phosphate buffered saline (PBS) (pH 7.4), and resuspended in 500 ml of binding buffer, into which 5 ml of fluorescein isothiocyanate (FITC)-labeled annexin V (Annexin V-FITC) and propidium iodide (PI) were added.…”

“…Therefore, novel agents that are effective and less toxic are urgently required to cure patients with multiple myeloma. It has been recently been reported that 2-aminophenoxazine-3-one (Phx-3), a most simple phenoxazine, which was originally found as questiomycin A produced by a streptomyces isolated from the soil in Tokyo, 3) and can now be synthesized by the reactions of oaminophenol with bovine hemoglobin solution, 4) shows in vitro anticancer acitivity against various cancer cells such as human melanoma, 5) human pancreatic cancer, 6) neuroblastoma, 7) and glioma 8) and against a mouse melanoma cell line, in vitro and in vivo. 9) Moreover, Phx-3 exerted apoptogentic activity against these cancer cell lines.…”

Inhibitory Effects of 2-Amino-3H-phenoxazin-3-one on the Melanogenesis of Murine B16 Melanoma Cell Line

“…13) In addition, APO can be synthesized from o-aminophenol by human erythrocytes. 14) APO has also been found to exert various biological effects, including anticancer, 15) antiviral, 16) and antichlamydia activities.…”

Anticancer activity of phenoxazines produced by bovine erythrocytes on colon cancer cells

“…Phx-1 and Phx-3 prepared by the conventional method using bovine hemoglobin solution (11) have been demonstrated to exert anticancer effects against a variety of cancer cells, including malignant melanoma cells (6), gastric cancer cells (7), neuroblastoma cells (8), multiple myeloma cells (9) and pancreatic cancer cells (23), though the anticancer activity of Phx-3 was much stronger than that of Phx-1 in these cancer cells. Thus, we studied the anticancer effects of Phx-1 and Phx-3 prepared by the present methods using bovine erythrocyte suspension on colon cancer cell lines COLO201, DLD1 and PMCO1 in vitro.…”