重楼皂苷Ⅰ通过p53信号通路诱导慢性粒细胞白血病细胞铁死亡

doi:10.20223/j.cnki.1000-8535.2025.05.005

摘要/Abstract

摘要： 目的探讨重楼皂苷Ⅰ（PPI）对慢性髓系白血病细胞（K562）细胞的抑制作用及可能的作用机制。方法采用CCK-8法筛选药物最适浓度,将培养时间为24 h的药物最适浓度作为后续实验的干预浓度。分组如下：（1）空白组;（2）PPI组;（3）抑制剂组;（4）PPI+抑制剂组。采用CCK-8法检测细胞增殖率;AO/EB染色观察细胞形态;流式细胞术检测凋亡率;ROS检测试剂盒检测活性氧（ROS）含量、还原型谷胱甘肽含量检测试剂盒检测谷胱甘肽（GSH）含量、细胞亚铁比色法测试盒检测细胞亚铁（Fe²⁺）含量;qRT-PCR法和蛋白免疫印迹法检测各组肿瘤蛋白53（p53）、钠氯离子依赖性氨基酸转运蛋白11（SLC7A11）、谷胱甘肽过氧化物酶4（GPX4）mRNA及蛋白表达量。结果 PPI抑制K562细胞的生长,且呈一定的剂量及时间依赖性（与同时间段的对照组0μmol/L比较,均P<0.01）。与空白组相比,PPI抑制K562细胞的增殖,提高了凋亡率,而铁死亡抑制剂（Ferrostian-1）的使用逆转了PPI对K562凋亡的促进作用（P<0.01）。与空白组相比,PPI组ROS、Fe²⁺含量升高,GSH含量下降,而铁死亡抑制剂的使用可下调ROS、Fe²⁺,上调GSH的含量（P<0.01）。PPI组较空白组p53 mRNA和蛋白表达水平升高,而SLC7A11、GPX4 mRNA和蛋白表达水平下降（P<0.05）;PPI+抑制剂组细胞较重楼皂苷组p53 mRNA和蛋白表达水平下降,而SLC7A11、GPX4 mRNA和蛋白表达水平升高（P<0.05）。结论 PPI能够有效抑制K562细胞增殖,促进K562细胞铁死亡,其分子机制可能与p53信号通路的调控有关。

关键词: 重楼皂苷Ⅰ, p53信号通路, 铁死亡, K562细胞

Abstract: Objective To investigate the inhibitory effect of polyphyllin I（PPI）on chronic myeloid leukemia cells（K562）and its possible mechanism．Methods K562 cell line was cultured in suitable environment,and the optimal concentration of the drug was screened by CCK-8 method．The optimal concentration of the drug cultured for 24 hours was used as the intervention concentration of the follow-up experiment．Cells were divided into the following groups：（1）blank group,（2）saponins group,（3）inhibitor group and（4）saponins + inhibitor group．The cell proliferation rate was detected by CCK-8 method．The cell morphology was observed by AO/EB staining．The apoptosis rate was detected by flow cytometry．The contents of reactive oxygen species（ROS）,glutathione（GSH）and ferrous（Fe²⁺）in different groups were detected,and the expression of mRNA and protein in different groups were detected by qRT- PCR and Western blot respectively．Results PPI significantly inhibited the growth of K562 cells in a dose-and time-dependent manner．Compared with the blank group,PPI significantly inhibited the proliferation of K562 cells and increased the apoptosis rate of K562 cells,while the use of ferroptosis inhibitor（Ferrostian-1）reversed the promoting effect of PPI on apoptosis of K562 cells．Compared with the blank group,the contents of reactive oxygen species（ROS）and ferrous iron（Fe²⁺）increased and the content of glutathione（GSH）decreased in the saponins group．The use of Ferrostian-1 could down-regulate the contents of ROS and Fe²⁺ and increase the content of GSH in the cells treated with the drug．Compared with the blank group,the expression of p53 mRNA and protein in the saponins group increased,while the expression of SLC7A11,GPX4 mRNA and protein decreased．The expression of p53 mRNA and protein in the saponins + inhibitor group was lower than that in the saponins group,while the expression levels of SLC7A11,GPX4 mRNA and protein increased．Conclusions PPI can effectively inhibit the proliferation of K562 cells and promote ferroptosis in K562 cells．The molecular mechanism can be related to the regulation of p53 signal pathway．

Key words: polyphyllin I, p53 signaling pathways, ferroptosis, K562 cells

陈冠君, 李建厂, 孙维梅, 周明轩, 尼坤洋, 王浩. 重楼皂苷Ⅰ通过p53信号通路诱导慢性粒细胞白血病细胞铁死亡[J]. 广州医药, 2025, 56(5): 605-615.

CHEN Guanjun, LI Jianchang, SUN Weimei, ZHOU Mingxuan, NI Kunyang, WANG Hao. Ferroptosis of chronic myeloid leukemia cells induced by polyphyllin I through p53 signal pathway[J]. Guangzhou Medical Journal, 2025, 56(5): 605-615.

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