Artesunate inhibits proliferation and promotes apoptosis and autophagy of nephroblastoma cell line SK-NEP-1

Objective To investigate the effects of artesunate (Art) on the proliferation, apoptosis, and autophagy of nephroblastoma cell line (SK-NEP-1). Methods SK-NEP-1 cells were intervened with different concentrations of Art (0, 10, 20, 40 and 80 μmol/L), and MTT method was applied to calculate the cell proliferation inhibition rate to screen the optimal intervention concentration; SK-NEP-1 cells were separated into control group, Art group, 3-MA group (Art+autophagy inhibitor, 3-methyladenine), and Rapa group (Art+autophagy activator rapamycin). EdU and flow cytometry were applied to detect cell proliferation and apoptosis, respectively; MDC staining was applied to detect autophagy in cells; the level of reactive oxygen species (ROS) in cells was detected by DCFH-DA fluorescent probe; the expression of proliferating cell nuclear antigen (PCNA), anti apoptotic factor B cell lymphomatoma-2 (Bcl-2), Bcl-2 associated X protein (Bax), microtubule junction protein 1 light chain 3Ⅱ/3Ⅰ (LC3Ⅱ/LC3Ⅰ), selective autophagy junction protein 1 (p62), and benzyl chloride 1 (Beclin-1) proteins in cells were detected by Western blot. Results Compared with 0 μmol/L Art, the proliferation inhibition rate of SK-NEP-1 cells was gradually increased after 10, 20, 40 and 80 μmol/L Art treatment (P＜0.05), and the IC50 value was 46.881 μmol/L, so 40 μmol/L Art was selected for follow-up experiments. Compared with the control group, the apoptosis rate, relative autophagy fluorescence intensity, ROS level, Bax, LC3 Ⅱ/LC3 Ⅰ, Beclin-1, PINK1, and Parkin protein expression levels of SK-NEP-1 cells in the Art group were obviously increased, the EdU positive cell rate, PCNA, Bcl-2, and P62 protein expression levels were obviously reduced (P＜0.05); The autophagy inhibitor 3-MA inhibited the promoting effect of Art on apoptosis and autophagy of nephroblastoma cells and inhibit proliferation (P＜0.05). Conclusions Art inhibits the proliferation of nephroblastoma cell line SK-NEP-1, and promotes autophagy and apoptosis.