Daphnoretin induces reactive oxygen species-mediated apoptosis in melanoma cells

Hui Wang; Xue Hu; Minjing Li; Zhaohai Pan; Defang Li; Qiusheng Zheng

doi:10.3892/ol.2021.12714

Daphnoretin induces reactive oxygen species-mediated apoptosis in melanoma cells

Oncol Lett. 2021 Jun;21(6):453. doi: 10.3892/ol.2021.12714. Epub 2021 Apr 8.

Authors

Hui Wang¹, Xue Hu², Minjing Li², Zhaohai Pan², Defang Li², Qiusheng Zheng^{1

2}

Affiliations

¹ Key Laboratory of Xinjiang Phytomedicine Resource and Utilization, Ministry of Education, School of Pharmacy, Shihezi University, Shihezi, Xinjiang 832003, P.R. China.
² Yantai Key Laboratory of Pharmacology of Traditional Chinese Medicine in Tumor Metabolism, School of Integrated Traditional Chinese and Western Medicine, Binzhou Medical University, Yantai, Shandong 264003, P.R. China.

Abstract

Research suggests that daphnoretin exhibits a diverse array of antitumor mechanisms and pharmacological activities. However, there is no definitive explanation for the antitumor mechanisms of daphnoretin in malignant melanoma. In the present study, MTT and colony formation assays demonstrated that daphnoretin significantly inhibited the proliferation of melanoma A375 and B16 cells. Following treatment with daphnoretin, apoptotic bodies were observed in A375 and B16 cells via Hoechst 33258 staining. Furthermore, western blot analysis revealed that the apoptosis-related proteins cleaved caspase-3, cleaved caspase-9, Bax, cytochrome c and apoptotic protease-activating factor 1 were significantly upregulated, while the expression levels of caspase-3, caspase-9 and Bcl-2 were downregulated in A375 and B16 cells. Flow cytometry and fluorescence microscopy revealed that daphnoretin induced higher levels of reactive oxygen species (ROS). Therefore, the results of the present study indicated that daphnoretin induced ROS-mediated mitochondria apoptosis in human (A375) and murine (B16) malignant melanoma cells.

Keywords: A375 cells; B16 cells; apoptosis; daphnoretin.

Grants and funding

The present study was funded by the National Natural Science Foundation of China (grant no. 31870338), the Key Research and Development Program of Shandong Province of China (grant no. 2019GSF108214), Taishan Scholars Construction Engineering of Shandong Province (grant no. tsqn201812099), the Dominant Disciplines Talent Team Development Scheme of Higher Education of Shandong Province (grant no. 2016052410) and the Introduction and Cultivation Project for Young Creative Talents of Higher Education of Shandong Province (grant no. 20191008198).