1.Inhibitory effect of capsaicin on B16-F10 melanoma cell migration via the phosphatidylinositol 3-kinase/Akt/Rac1 signal pathway.
Dong Hoon SHIN ; Ok Hee KIM ; Hye Seung JUN ; Mi Kyung KANG
Experimental & Molecular Medicine 2008;40(5):486-494
Capsaicin (trans-8-methyl-N-vanillyl-6-nonenamide), the major pungent ingredient of red pepper, has been reported to possess anti-carcinogenic and anti-mutagenic activities. In this study, the anti-migration activity of capsaicin on highly metastatic B16-F10 melanoma cells was investigated. Capsaicin significantly inhibited the migration of melanoma cells without showing obvious cellular cytotoxicity at low doses. This effect correlated with the down-regulation of phosphatidylinositol 3-kinase (PI3-K) and its downstream target, Akt. Although B16-F10 cell migration was increased by the PI3-K activator through the activation of Akt, these PI3-K activator-induced phenomena were attenuated by capsaicin. Moreover, capsaicin was found to significantly inhibit Rac1 activity in a pull-down assay. These results demonstrate that capsaicin inhibits the migration of B16-F10 cells through the inhibition of the PI3-K/Akt/Rac1 signal pathway. The present investigation suggests that capsaicin targets PI3-K/Akt/ Rac1-mediated cellular events in B16-F10 melanoma cells. Consequently, capsaicin administration should be considered an effective approach for the suppression of invasion and metastasis in malignant melanoma chemotherapy.
1-Phosphatidylinositol 3-Kinase/*metabolism
;
Animals
;
Capsaicin/*pharmacology
;
Cell Line, Tumor
;
Cell Movement/*drug effects
;
Cell Survival/drug effects
;
Dose-Response Relationship, Drug
;
Immunoblotting
;
Melanoma, Experimental/metabolism/pathology/physiopathology
;
Mice
;
Proto-Oncogene Proteins c-akt/*metabolism
;
Signal Transduction/*drug effects
;
rac1 GTP-Binding Protein/*metabolism
2.Introduction of the CIITA gene into tumor cells produces exosomes with enhanced anti-tumor effects.
Yeong Shin LEE ; Soo Hyun KIM ; Jung Ah CHO ; Chul Woo KIM
Experimental & Molecular Medicine 2011;43(5):281-290
Exosomes are small membrane vesicles secreted from various types of cells. Tumor-derived exosomes contain MHC class I molecules and tumor-specific antigens, receiving attention as a potential cancer vaccine. For induction of efficient anti-tumor immunity, CD4+ helper T cells are required, which recognize appropriate MHC class II-peptide complexes. In this study, we have established an MHC class II molecule-expressing B16F1 murine melanoma cell line (B16F1-CIITA) by transduction of the CIITA (Class II transactivator) gene. Exosomes from B16-CII cells (CIITA-Exo) contained a high amount of MHC class II as well as a tumor antigen TRP2. When loaded on dendritic cells (DCs), CIITA-Exo induced the increased expression of MHC class II molecules and CD86 than the exosomes from the parental cells (Exo). In vitro assays using co-culture of immunized splenocytes and exosome-loaded DCs demonstrated that CIITA-Exo enhanced the splenocyte proliferation and IL-2 secretion. Consistently, compared to B16-Exo, CIITA-Exo induced the increased mRNA levels of inflammatory cytokines such as TNF-alpha, chemokine receptor CCR7 and the production of Th1-polarizing cytokine IL-12. A tumor preventive model showed that CIITA-Exo significantly inhibited tumor growth in a dose-dependent manner. Ex vivo assays using immunized mice demonstrated that CIITA-Exo induced a higher amount of Th1-polarized immune responses such as Th1-type IgG2a antibodies and IFN-gamma cytokine as well as TRP2-specific CD8+ T cells. A tumor therapeutic model delayed effects of tumor growth by CIITA-Exo. These findings indicate that CIITA-Exo are more efficient as compared to parental Exo to induce anti-tumor immune responses, suggesting a potential role of MHC class II-containing tumor exosomes as an efficient cancer vaccine.
Animals
;
Cancer Vaccines/genetics/immunology
;
Cell Line, Tumor
;
Cell Proliferation
;
Dendritic Cells/immunology
;
Exosomes/genetics/*metabolism
;
Gene Expression Regulation
;
Gene Transfer Techniques
;
Immunity, Cellular/immunology
;
Immunity, Humoral/immunology
;
Immunotherapy
;
Lymphocyte Activation/immunology
;
Melanoma, Experimental/mortality/pathology/*physiopathology
;
Mice
;
Mice, Inbred C57BL
;
Nuclear Proteins/*genetics/*metabolism
;
Survival Analysis
;
T-Lymphocytes/immunology/metabolism
;
Trans-Activators/*genetics/*metabolism
;
Transduction, Genetic
3.IL-1beta Acts in Synergy with Endogenous IL-1beta in A375-S2 Human Melanoma Cell Apoptosis Through Mitochondrial Pathway.
Che WANG ; Min Wei WANG ; Shin Ichi TASHIRO ; Satoshi ONODERA ; Takashi IKEJIMA
Journal of Korean Medical Science 2005;20(4):555-561
Interleukin-1beta (IL-1beta) is a pivotal proinflammatory cytokine. To investigate the mechanism of IL-1beta-induced cell death in human malignant melanoma A375-S2 cells, MTT assay, photomicroscopical observation, DNA agarose gel electrophoresis, radioimmunoassay and Western blot analysis were carried out. IL-1beta did not only induce nuclear condensation and DNA fragmentation, but also increased degradation of two substrates of caspase-3, poly ADP-ribose polymerase (PARP) and inhibitor of caspase-activated DNase (ICAD). Simultaneously, release of precursor of IL-1beta (pro-IL-1beta) and endogenous IL-1beta production were involved in the apoptotic process. IL-1beta enhanced the ratio of Bax/Bcl-2 and Bax/Bcl-xL expression and up-regulated apoptosis inducing factor (AIF) expression, which required the activation of downstream caspases. These results suggest that IL-1beta induces endogenous IL-1beta production, enhances cleavage of caspase downstream substrates and promotes mitochondria mediated apoptosis in A375-S2 cells.
Apoptosis/*drug effects
;
Blotting, Western
;
Caspase 1/metabolism
;
Caspases/metabolism
;
Cell Line, Tumor
;
Cell Proliferation/drug effects
;
Cell Survival/drug effects
;
Comparative Study
;
DNA Fragmentation/drug effects
;
Deoxyribonucleases/metabolism
;
Dose-Response Relationship, Drug
;
Enzyme Activation/drug effects
;
Humans
;
Interleukin-1/biosynthesis/*pharmacology/physiology
;
Interleukin-6/pharmacology
;
Lymphotoxin/pharmacology
;
Melanoma/metabolism/pathology/physiopathology
;
Mitochondria/*physiology
;
Poly(ADP-ribose) Polymerases/metabolism
;
Proto-Oncogene Proteins c-bcl-2/biosynthesis
;
Time Factors