Objective: Ranunculus japonicus thumb. has been used as a traditional medicine for the treatment of asthma, malaria, tuberculosis, hepatitis, jaundice and so on. The aim of this study was to investigate the anti-cancer effect and action mechanism of the petroleum ether extract of Ranunculus japonicus Thumb. (RJTPEE) on human neuroblastoma SH-SY5Y cells. Methods: Cell viability was detected using MTT assay. FCM and Hoechst33258 staining was used to examine the cell cycle as well as apoptosis, and in addition, Western blot was also used to examine the expression of a range of related proteins. Results: MTT assay showed that RJTPEE could inhibit SH-SY5Y cells proliferation in concentration- and time-dependent manners. FCM analysis showed that RJTPEE could induce S and G2/M phase arrest, cell apoptosis, cell autophagy, the mitochondrial membrane potential loss (ΔΨm) and increase the production of intracellular ROS of SH-SY5Y cells. Topical morphological changes of apoptotic body formation were also observed by Hoechst 33258 staining. Western blot analysis indicated that the expressions of cell cycle-related protein Cyclin B1/D1 and CDK1/2/4/6 were obviously decreased. Meanwhile, obvious increase of apoptosis-related protein expressions, such as Bax, cytochrome c, caspase-3, caspase-9, PARP-1, and decrease of Bcl-2 protein expression were observed. Furthermore, Akt/MAPKs pathway-related proteins were also detected. Moreover, the decreased expression of autophagy-related proteins LC3 I protein indicated that RJTPEE could also enhance the autophagy ability of cells. Conclusion: RJTPEE could exert the antitumor activity on SH-SY5Y cells by inducing cell cycle arrest, cell autophagy, the mitochondrion-dependent cell apoptosis and targeting the Akt/MAPKs signaling pathway. RJTPEE has a promising therapeutic potential for neuroblastoma.

Jie Ren, Jichao Zhou, Wenbin Wang, Qun Wang, Wenjing Huang, Kun Hu. Ranunculus Japonicus Induces Cell Cycle Arrest, Apoptosis, Autophagy and Targeting the Akt/Mapks Pathways in Human Neuroblastoma SH-SY5Y Cells. International Journal of Frontiers in Medicine (2022), Vol. 4, Issue 3: 24-33. https://doi.org/10.25236/IJFM.2022.040305.

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