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Chinese Journal of Clinical Pharmacology and Therapeutics ›› 2025, Vol. 30 ›› Issue (1): 51-60.doi: 10.12092/j.issn.1009-2501.2025.01.006

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Effects and mechanism of ethanol extract of Angelica sinensis (Oliv.) Diels on cell proliferation inhibition and apoptosis induction in B16-F10 melanoma cells

KE Jiajie1, SHEN Yuzhou1, XU Yaping2, CHEN Yupei1, CHEN Peiyuan3, WU Hongtan1   

  1. 1 School of Public Health and Medical Technology, Xiamen Medical College, Xiamen 361023, Fujian, China; 2 School of Basic Medicine, Xiamen Medical College, Xiamen 361023, Fujian, China; 3 Department of Dermatology, Xiamen University Affiliated Chenggong Hospital, Xiamen 361003, Fujian, China
  • Received:2023-12-13 Revised:2024-07-10 Online:2025-01-26 Published:2025-01-02

Abstract:

AIM: To explore the effect and mechanism of ethanol extract of Angelica sinensis?(Oliv.) Diels (EEA) on cell proliferation and apoptosis in B16-F10 melanoma cells. METHODS: Cell viability was analyzed by MTT method. Cell proliferation was detected by colony formation assay. The inverted microscope was used to observe the changes of cell growth confluence and morphology. Hoechst 33342 staining was used to detect cell apoptosis. Flow cytometry (FCM) was used to detect cell cycle and apoptosis. Transmission electron microscopy (TEM) was used to observe the changes of cell mitochondrial structure. Western blot was used to detect the expression levels of cell cycle, apoptosis, mitochondrial biogenesis, and mitochondrial dynamics-related proteins. RESULTS: Compared with the blank control group, the cell viability of B16-F10 melanoma cells was reduced after EEA (10-400 μg/mL) treatment for 24 h and 48 h, respectively (P<0.05, P<0.01). The decreased cell growth confluence, morphological changes such as shrinkage, rounding, and reduction in the volume, and apoptotic?morphologic?changes such as chromatin condensation were observed after EEA (100 μg/mL and 200 μg/mL) treatment for 24 h. The number of cell clones was decreased after EEA (10-200 μg/mL) treatment for 14 d (P<0.01). The morphology of mitochondria became more round and shorter, and the inner mitochondrial matrices were either damaged or absent after 200 μg/mL EEA treatment for 24 h. The ratio of cells in G0/G1 phase and the early apoptosis rate of cells were higher than those of the blank control group (P<0.01) after EEA (20-200 μg/mL) treatment for 24 h. Western blot results showed that compared with the blank control group, the protein expression levels of cleaved caspase-9, Bax, DRP1, and FIS1 were up-regulated (P<0.05, P<0.01), and the protein expression levels of cyclin D1, cyclin E, CDK2, CDK4, Bcl-2, Bad, Bcl-XL, SIRT1, PGC-1α, NRF1, TFAM, MFN2, and OPA1 were down-regulated (P<0.05, P<0.01). CONCLUSION: EEA has an inhibitory effect on the proliferation of B16-F10 melanoma cells, which may be related to the induction of G1/S cell cycle arrest and mitochondrial apoptotic pathway, and the disruption of mitochondrial biogenesis and mitochondrial dynamics.

Key words: EEA, B16-F10 melanoma cells, cell cycle, apoptosis, mitochondrial biogenesis and mitochondrial dynamics

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