Abstract

Norcantharidin (NCTD), a demethylated derivative of cantharidin, has been reported to exhibit activity against various types of cancers. However, the anti-invasive effects of NCTD and its molecular mechanism in human mucoepidermoid carcinoma (MEC) remain incompletely elucidated. Clonogenic, wound healing, invasion, zymography, western blotting and immunocytochemistry assays were performed in YD-15 cells to investigate the anti-invasive effect of NCTD and its molecular mechanism of action. The inhibitory effects of NCTD on invasiveness were compared with those of a novel focal adhesion kinase (FAK) kinase inhibitor, PF-562271. NCTD markedly suppressed the colony formation, migration, and invasion of YD-15 cells as well as the activities of MMP-2 and MMP-9. It disrupted F-actin reorganization through suppressing the FAK/Paxillin axis. Moreover, NCTD exhibited a powerful anti-invasive effect compared with that of PF-562271 in YD-15 cells. Collectively, these results suggest that NCTD has a potential anti-invasive activity against YD-15 cells. This study may clarify the impact of NCTD on migration and invasion of human MEC cells.

Highlights

  • Mucoepidermoid carcinoma (MEC) is the most common malignant tumor of the major salivary glands and histologically comprises variable mixtures of mucin-secreting, intermediated, and epidermoid cells [1,2]

  • We we investigated the anti-invasive potential of NCTD in the YD-15 human MEC cell line and compared investigated the anti-invasive potential of NCTD in the YD-15 human MEC cell line and compared its its anti-cancer function with that of PF562271, a potent focal adhesion kinase (FAK) inhibitor

  • NCTD-treated YD-15 cells increased (Supplementary Figure S1C,D). These results suggest that low NCTD-treated YD-15 cells increased (Supplementary Figure S1C,D)

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Summary

Introduction

Mucoepidermoid carcinoma (MEC) is the most common malignant tumor of the major salivary glands and histologically comprises variable mixtures of mucin-secreting, intermediated, and epidermoid cells [1,2]. Molecules 2019, 24, 1928 kinase (FAK) is a non-receptor tyrosine kinase that acts as an important mediator of integrinmediated signaling between cells and the ECM. It serves as a scaffolding protein for the binding sites signaling between cells and the ECM. The functional significance of FAK activity in MEC has not yet MEC has not yet been fully understood Natural products such as plant and animal-derived drugs play an increasingly important role in Natural products such as plant and animal-derived drugs play an increasingly important role in cancer treatment due to their low side-effects and high efficacy [15,16]. Anti-cancer function with that of PF562271, a potent FAK inhibitor

Results
NCTD Inhibits the Invasiveness of YD-15 Cells More Potently than PF562271
Discussion
Cell Culture and Chemical Treatment
Trypan Blue Exclusion Assay
Clonogenic Formation Assay
Scratch Wound Healing Assay
Matrigel Invasion Assay
Gelatin Zymography
F-Actin Staining Using TRITC-Conjugated Phalloidin
Western Blot Analysis
Statistical Analysis
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