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Suppression of the migration and invasion is mediated by triptolide in B16F10 mouse melanoma cells through the
Hui-Yu Jao1, Fu-Shun Yu2, Chun-Shu Yu3
1Department of Biological Science and Technology, China Medical University, Taichung, 404, Taiwan, ROC.
Abstract:
Melanoma cancer is one of the major causes of death in humans worldwide. Triptolide is one of the active components of Tripterygium wilfordii Hook F, and has biological activities including induced cell cycle arrest and induction of apoptosis but its antimetastatic effects on murine melanoma cells have not yet been elucidated. Herein, we investigated the effect of triptolide on the inhibition of migration and invasion and possible associated signal pathways in B16F10 murine melanoma cancer cells. Wound healing assay and Matrigel Cell Migration Assay and Invasion System demonstrated that triptolide marked inhibiting the migration and invasion of B16F10 cells. Gelatin zymography assay demonstrated that triptolide significantly inhibited the activities of matrix metalloproteinases-2 (MMP-2). Western blotting showed that triptolide markedly reduced CXCR4, SOS1, GRB2, p-ERK, FAK, p-AKT, Rho A, p-JNK, NF-κB, MMP-9, and MMP-2 but increased PI3K and p-p38 and COX2 after compared to the untreated (control) cells. Real time PCR indicated that triptolide inhibited the gene expression of MMP-2, FAK, ROCK-1, and NF-κB but did not significantly affect TIMP-1 and -2 gene expression in B16F10 cells in vitro. EMSA assay also showed that triptolide inhibited NF-κB DNA binding in a dose-dependent manner. Confocal laser microscopy examination also confirmed that triptolide inhibited the expression of NF-κB in B16F10 cells. Taken together, we suggest that triptolide inhibited B16F10 cell migration and invasion via the inhibition of NF-κB expression then led to suppress MMP-2 and -9 expressions. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1974-1984, 2016.
Insights
Triptolide effectively inhibits melanoma cell migration and invasion by suppressing NF-κB expression, leading to reduced matrix metalloproteinase activity. This study highlights triptolide
Area of Science:
- Oncology
- Molecular Biology
- Pharmacology
Background:
- Melanoma is a significant global health concern, causing numerous deaths worldwide.
- Triptolide, derived from Tripterygium wilfordii Hook F, exhibits known cytotoxic properties like cell cycle arrest and apoptosis induction.
- The antimetastatic potential of triptolide in melanoma, particularly its effects on cell migration and invasion, remains largely unexplored.
Purpose of the Study:
- To investigate the antimetastatic effects of triptolide on B16F10 murine melanoma cells.
- To elucidate the potential molecular mechanisms and signaling pathways involved in triptolide's antimetastatic activity.
Main Methods:
- Cell migration and invasion assays (wound healing, Matrigel assay).
- Enzyme activity analysis (Gelatin zymography).
- Protein expression analysis (Western blotting, confocal microscopy).
- Gene expression analysis (Real-time PCR).
- DNA binding assay (EMSA).
Main Results:
- Triptolide significantly inhibited the migration and invasion of B16F10 melanoma cells.
- Triptolide suppressed the activity and expression of matrix metalloproteinases (MMP-2 and MMP-9).
- Triptolide reduced the expression of key signaling molecules including NF-κB, FAK, CXCR4, and ERK, while affecting PI3K/AKT and MAPK pathways.
- Triptolide inhibited NF-κB DNA binding and expression in a dose-dependent manner.
Conclusions:
- Triptolide demonstrates potent antimetastatic effects against B16F10 murine melanoma cells.
- The mechanism involves the inhibition of NF-κB signaling, subsequently suppressing MMP-2 and MMP-9 expression.
- Triptolide represents a potential therapeutic agent for melanoma treatment, warranting further investigation.
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