Licochalcone�D induces apoptosis and inhibits migration and invasion in human melanoma A375 cells

Abstract

The aim of the present study was to determine the effects of Licochalcone D (LD) on the apoptosis and migration and invasion in human melanoma A375 cells. Cell proliferation was determined by sulforhodamine B assay. Apoptosis was assessed by Hoechst 33258 and Annexin V-FITC/PI staining and JC-1 assay. Total intracellular reactive oxygen species (ROS) was examined by DCFH-DA. Wound healing and Transwell assays were used to detect migration and invasion of the cells. The activities of matrix metalloproteinase (MMP-2 and MMP-9) were assessed via gelatin zymography. Tumor growth in vivo was evaluated in C57BL/6 mice. RT-PCR, qPCR, ELISA and western blot analysis were utilized to measure the mRNA and protein levels. Our results showed that LD inhibited the proliferation of A375 and SK-MEL-5 cells in a concentration-dependent manner. After treatment with LD, A375 cells displayed obvious apoptotic characteristics, and the number of apoptotic cells was significantly increased. Pro-apoptotic protein Bax, caspase-9 and caspase-3 were upregulated, while anti-apoptotic protein Bcl-2 was downregulated in the LD-treated cells. Meanwhile, LD induced the loss of mitochondrial membrane potential (ΔΨm) and increased the level of ROS. ROS production was inhibited by the co-treatment of LD and free radical scavenger N-acetyl-cysteine (NAC). Furthermore, LD also blocked A375 cell migration and invasion in vitro which was associated with the downregulation of MMP-9 and MMP-2. Finally, intragastric administration of LD suppressed tumor growth in the mouse xenograft model of murine melanoma B16F0 cells. These results suggest that LD may be a potential drug for human melanoma treatment by inhibiting proliferation, inducing apoptosis via the mitochondrial pathway and blocking cell migration and invasion.

HIF‐1α/VEGF signaling‐mediated epithelial–mesenchymal transition and angiogenesis is critically involved in anti‐metastasis effect of luteolin in melanoma cells

“…Cell migration was measured by wound‐healing assay to evaluate the inhibit migration effect of luteolin in vitro (Si et al, ). A375 and B16‐F10 cells were seeded in 60‐mm dishes at the density of 8 × 10 5 cells per dish to 90–100% confluence.…”

“…Cell invasion was detected using Matrigel‐coated transwell chambers (8 μm pore size; Corning, USA) assay according to the manufacturer's instructions (Si et al, ). After pretreatment with or without (control) various concentrations (5, 10, and 20 μM) of luteolin for 24 hr, A375 and B16‐F10 cells were harvested and seeded into the upper chamber at the density of 4 × 10 4 cells per well in serum free medium.…”

Licochalcone D Induces ROS-Dependent Apoptosis in Gefitinib-Sensitive or Resistant Lung Cancer Cells by Targeting EGFR and MET

“…Various pharmacological actions, including anti-oxidant, anti-biotic, anti-ulcer, and anti-carcinogenic effects, have been described for LCD [2]. Previous studies have demonstrated that LCD can induce cell apoptosis and suppress migration and invasion in skin cancer [3]. In addition, LCD can suppress the proliferation in oral cancer cells [4].…”

Natural Chalcones in Chinese Materia Medica: Licorice

“…In addition, it attenuated migration, adhesion, and invasion of cancer cells accompanied with the downregulated protein expression of MMP-9 and nuclear translocation of nuclear factor-κB (NF-κB) [115]. Similar to licochalcone B, licochalcones C and D induced apoptosis by altering the expression of Bcl-2 family member genes and activating the caspase-mediated cell death signaling pathway [45,46]. Meanwhile, licochalcone D induced apoptosis through mitochondrial pathway and blocked cell migration and invasion into surrounding tissues by reducing the activity and expression of MMP-2 and MMP-9 [46].…”

“…Similar to licochalcone B, licochalcones C and D induced apoptosis by altering the expression of Bcl-2 family member genes and activating the caspase-mediated cell death signaling pathway [45,46]. Meanwhile, licochalcone D induced apoptosis through mitochondrial pathway and blocked cell migration and invasion into surrounding tissues by reducing the activity and expression of MMP-2 and MMP-9 [46]. In addition to increasing the expression of proteins (proapoptotic factors, caspase-3, and PARP), suppressing the constitutive NF-κB activation, and downregulating Bcl-2 and Bax, licochalcone E reduced tumor growth and metastasis through inhibiting tube formation of vascular endothelial cells [47][48][49].…”