青蒿琥酯抑制肾母细胞瘤细胞系SK-NEP-1增殖和促进凋亡与自噬

doi:10.16352/j.issn.1001-6325.2025.04.0493

基础医学与临床 ›› 2025, Vol. 45 ›› Issue (4): 493-498.doi: 10.16352/j.issn.1001-6325.2025.04.0493

青蒿琥酯抑制肾母细胞瘤细胞系SK-NEP-1增殖和促进凋亡与自噬

魏建新¹, 房彦乐², 鲁雨博³, 高宇光¹, 郎兴^1*, 李静涛¹, 马新生¹

1.邯郸市中心医院小儿外二科,河北邯郸 056000;
2.河北医科大学第二医院普外三科,河北石家庄 050000
3.邯郸市中心医院药学部,河北邯郸 056000

收稿日期:2024-07-25 修回日期:2024-10-30 出版日期:2025-04-05 发布日期:2025-03-24
通讯作者: ^*swsus0@163.com
基金资助:
河北省2024年度中医药类科学研究课题计划项目(2024195)

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

WEI Jianxin¹, FANG Yanle², LU Yubo³, GAO Yuguang¹, LANG Xing^1*, LI Jingtao¹, MA Xinsheng¹

1. the Second Department of Pediatric Surgery, Handan Central Hospital, Handan 056000;
2. the Third Department of General Surgery, the Second Hospital of Hebei Medical University, Shijiazhuang 050000;
3. Department of Pharmacy, Handan Central Hospital, Handan 056000, China

Received:2024-07-25 Revised:2024-10-30 Online:2025-04-05 Published:2025-03-24

摘要/Abstract

摘要： 目的　探讨青蒿琥酯(Art)对肾母细胞瘤细胞系(SK-NEP-1)增殖、凋亡和自噬的影响。方法　以不同浓度的Art(0、10、20、40和80 μmol/L)干预SK-NEP-1细胞,MTT法检测细胞增殖抑制率,筛选最佳干预浓度;将SK-NEP-1细胞分为对照组、Art组、3-MA组(Art+自噬抑制剂,3-甲基腺嘌呤)、Rapa组(Art+自噬激活剂雷帕霉素),EdU、流式细胞测量术分别检测细胞增殖、凋亡;丹磺酰尸胺(MDC)染色法检测细胞自噬;DCFH-DA荧光探针检测细胞活性氧(ROS)的水平;Western blot检测细胞中增殖细胞核抗原(PCNA)、细胞抗凋亡因子B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、微管连接蛋白1轻链3Ⅱ/3Ⅰ(LC3Ⅱ/LC3Ⅰ)、选择性自噬接头蛋白1(p62)和Beclin-1蛋白表达。结果　与0 μmol/L Art相比,10、20、40和80 μmol/L Art处理后SK-NEP-1细胞增殖抑制率逐渐升高(P＜0.05),IC₅₀值为46.881 μmol/L,故选取40 μmol/L Art进行后续实验。与对照组相比,Art组SK-NEP-1细胞凋亡率、相对自噬荧光强度、ROS水平、Bax、LC3Ⅱ/LC3Ⅰ、Beclin-1、PINK1、Parkin蛋白表达水平显著升高,EdU阳性细胞率、PCNA、Bcl-2、p62蛋白表达水平显著降低(P＜0.05);自噬抑制剂3-MA可减弱Art对肾母细胞瘤细胞凋亡和自噬的促进作用,对增殖的抑制作用(P＜0.05)。结论　Art抑制肾母细胞瘤细胞系的增殖,促进细胞自噬和凋亡。

关键词: 青蒿琥酯, 肾母细胞瘤, 增殖, 凋亡, 自噬

Abstract: 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.

Key words: artesunate, nephroblastoma, proliferation, apoptosis, autophagy

中图分类号:

R737.11

魏建新, 房彦乐, 鲁雨博, 高宇光, 郎兴, 李静涛, 马新生. 青蒿琥酯抑制肾母细胞瘤细胞系SK-NEP-1增殖和促进凋亡与自噬[J]. 基础医学与临床, 2025, 45(4): 493-498.

WEI Jianxin, FANG Yanle, LU Yubo, GAO Yuguang, LANG Xing, LI Jingtao, MA Xinsheng. Artesunate inhibits proliferation and promotes apoptosis and autophagy of nephroblastoma cell line SK-NEP-1[J]. Basic & Clinical Medicine, 2025, 45(4): 493-498.

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青蒿琥酯抑制肾母细胞瘤细胞系SK-NEP-1增殖和促进凋亡与自噬

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

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