N -propionyl-4- S -cysteaminylphenol induces apoptosis in B16F1 cells and mediates tumor-specific T-cell immune responses in a mouse melanoma model

• Published in: Journal of dermatological science Vol. 67; no. 1; pp. 51 - 60
• Main Authors: Ishii-Osai, Yasue, Yamashita, Toshiharu, Tamura, Yasuaki, Sato, Noriyuki, Ito, Akira, Honda, Hiroyuki, Wakamatsu, Kazumasa, Ito, Shosuke, Nakayama, Eiichi, Okura, Masae, Jimbow, Kowichi
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Ireland Ltd 01.07.2012
• Subjects: Animals; Antibodies - administration & dosage; Antineoplastic Agents - pharmacology; Apoptosis; Apoptosis - drug effects; Caspase 3 - metabolism; CD4-Positive T-Lymphocytes - drug effects; CD4-Positive T-Lymphocytes - immunology; CD8-Positive T-Lymphocytes - drug effects; CD8-Positive T-Lymphocytes - immunology; Cell Line, Tumor; Cell Proliferation - drug effects; Cystamine - analogs & derivatives; Cystamine - pharmacology; Dermatology; DNA Fragmentation; Dose-Response Relationship, Drug; Enzyme Activation; Female; Flow Cytometry; Humans; Immunity, Cellular - drug effects; Intramolecular Oxidoreductases - metabolism; Melanins - biosynthesis; Melanogenesis; Melanoma, Experimental - drug therapy; Melanoma, Experimental - immunology; Melanoma, Experimental - metabolism; Melanoma, Experimental - pathology; Mice; Mice, Inbred C57BL; N-propionyl-4-S-cysteaminylphenol; NIH 3T3 Cells; Phenols - pharmacology; Reactive oxygen species; Reactive Oxygen Species - metabolism; Time Factors; Tumor Burden - drug effects; Tumor suppression; Tyrosinase-related protein; TYRP-1; NPr-4- S-CAP; heat shock protein; tyrosinase-related protein-2; phosphate-buffered saline; tyrosinase-related protein-1; TYRP-2; DMEM; Tumor suppression; FBS; reactive oxygen species; NPr-2- S-CAP; tumor necrosis factor-related apoptosis-inducing ligand; AMF; N-propionyl-2- S-cysteaminylphenol; PBS; TRAIL; HSP; N-propionyl-4- S-cysteaminylphenol; RPMI1640 medium; Melanogenesis; Dulbecco's modified Eagle's medium; fetal bovine serum; Tyrosinase-related protein; ROS; RPMI; alternating magnetic field; Apoptosis; Reactive oxygen species; N-propionyl-4-S-cysteaminylphenol; NPr-4-S-CAP; NPr-2-S-CAP
• ISSN: 0923-1811; 1873-569X
• DOI: 10.1016/j.jdermsci.2012.04.009

Abstract Background N -propionyl-4- S -cysteaminylphenol (NPr-4- S -CAP) is selectively incorporated into melanoma cells and degrades them. However, it remains unclear whether NPr-4- S -CAP can induce cell death associated with the induction of host immune responses and tumor suppression in vivo. Objective To examine the molecular mechanism of NPr-4- S -CAP-mediated cytotoxicity toward melanoma cells and to test whether NPr-4- S -CAP can suppress transplanted primary and secondary B16F1 melanomas. Methods Cytotoxicity and apoptosis of melanoma cells were assessed by cell counting, flow cytometry, and detection of reactive oxygen species (ROS) and apoptotic molecules. NPr-4- S -CAP-associated host immunity was studied using a B16F1 mouse melanoma model through the application of CD4- and CD8-specific antibodies and tetramer assay. Results NPr-4- S -CAP suppressed growth of pigmented melanoma cells associated with an increase of intracellular ROS, activation of caspase 3 and DNA fragmentation, suggesting that NPr-4- S -CAP mediated ROS production, eliciting apoptosis of melanoma cells. Growth of transplanted B16F1 melanomas was inhibited after the consecutive intratumoral injections of NPr-4- S -CAP, and the tumor growth after rechallenge of B16F1 was significantly suppressed in the treated mice. This suppression occurred when the treated mice were given the anti-CD4 antibody, but not the anti-CD8 antibody. Tetramer assay demonstrated increased TYRP-2-specific CD8+ T cells in the lymph node and spleen cells prepared from NPr-4- S -CAP-treated B16F1-bearing mice. Conclusions These suggest that NPr-4- S -CAP induces apoptosis in melanoma cells through ROS production and generates CD8+ cell immunity resulting in the suppression of rechallenged B16F1 melanoma.