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

Si,Lingling; Yan,Xinyan; Hao,Wenjin; Ma,Xiaoyi; Ren,Huanhuan; Ren,Boxue; Li,Defang; Dong,Zhengping; Zheng,Qiusheng

doi:10.3892/or.2018.6329

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

Authors:
- Lingling Si
- Xinyan Yan
- Wenjin Hao
- Xiaoyi Ma
- Huanhuan Ren
- Boxue Ren
- Defang Li
- Zhengping Dong
- Qiusheng Zheng
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Affiliations: School of Integrated Traditional Chinese and Western Medicine, Binzhou Medical University, Yantai, Shandong 264000, P.R. China, People's Hospital of Xinjiang Uygur Autonomous Region, Uygur, Xinjiang 830001, P.R. China, School of Pharmacy, Shihezi University, Key Laboratory of Xinjiang Endemic Phytomedicine Resources, Ministry of Education, Shihezi, Xinjiang 832002, P.R. China
Published online on: March 20, 2018 https://doi.org/10.3892/or.2018.6329
Pages: 2160-2170
Copyright: © Si et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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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.

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