iFSP1对SKOV3细胞铁死亡的影响及机制研究^*

Effects and mechanism of iFSP1 on ferroptosis in SKOV3 cells

摘要

目的 探究铁死亡抑制蛋白1抑制剂(iFSP1)对SKOV3细胞铁死亡的影响及机制, 为卵巢癌临床治疗策略提供新的理论依据。方法 使用光学显微镜观察iFSP1对细胞形态和生长状态的影响; 采用CCK-8比色法检测iFSP1单独处理及与铁死亡抑制剂Lip-1联合处理对细胞活力的影响; 利用活性氧(ROS)检测试剂盒分析细胞内ROS的水平; 使用Western blot检测iFSP1作用后细胞内铁死亡抑制蛋白1的表达情况; 采用CCK-8法检测铁死亡诱导剂RSL3、铁死亡诱导剂RSL3联合iFSP1对细胞活力的影响。结果 iFSP1可抑制SKOV3细胞增殖, 并且随着iFSP1浓度升高和作用时间的延长, 细胞抑制率逐步上升(P<0.05); 铁死亡抑制剂Lip-1可减弱iFSP1对SKOV3细胞的抑制效应(P<0.05); iFSP1可提升SKOV3细胞的ROS水平(P<0.05); iFSP1可下调SKOV3细胞的FSP1蛋白表达水平(P<0.05); iFSP1能够增强铁死亡诱导剂RSL3对SKOV3细胞的生长抑制(P<0.05)。结论 iFSP1可下调FSP1蛋白表达水平, 诱导SKOV3细胞铁死亡, 从而抑制SKOV3细胞的增殖。iFSP1可增强铁死亡诱导剂RSL3对SKOV3细胞的抑制效应。

Abstract

Objective To explore the impact and underlying mechanisms of ferroptosis suppressor protein 1 inhibitor (iFSP1) on ferroptosis in SKOV3 cells, aiming to provide a novel theoretical foundation for clinical treatment strategies of ovarian cancer. Methods The effects of iFSP1 on cell morphology and growth status were observed using optical microscopy. The CCK-8 assay was employed to evaluate the impact of iFSP1 alone or in combination with the ferroptosis inhibitor Lip-1 on cell viability. Intracellular levels of reactive oxygen species (ROS), were quantitatively analyzed using an ROS detection kit. Western blotting was performed to measure the expression levels of FSP1 protein in cells treated with iFSP1. Additionally, the CCK-8 assay was used to assess the effects of ferroptosis inducer RSL3 alone or in combination with iFSP1 on cell viability. Results iFSP1 significantly inhibited the proliferation of SKOV3 cells, with the inhibition rate increasing progressively with higher concentrations of iFSP1 and longer treatment durations (P<0.05). The ferroptosis inhibitor Lip-1 attenuated the inhibitory effect of iFSP1 on SKOV3 cells (P<0.05). iFSP1 markedly elevated ROS levels in the SKOV3 cells and downregulated the expression of FSP1 protein (P<0.05). Furthermore, iFSP1 enhanced the growth-inhibitory effect of the ferroptosis inducer RSL3 on SKOV3 cells (P<0.05). Conclusion iFSP1 induces ferroptosis in SKOV3 cells by downregulating FSP1 protein expression, thereby inhibiting SKOV3 cells proliferation. Additionally, iFSP1 enhances the inhibitory effect of the ferroptosis inducer RSL3 on SKOV3 cells.

关键词

iFSP1 / 铁死亡 / 卵巢癌 / RSL3 / 铁死亡抑制蛋白1

Key words

iFSP1 / Ferroptosis / Ovarian cancer / RSL3 / Ferroptosis suppressor protein 1

参考文献

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基金资助

^*吉林省科技发展计划项目(YDZJ202201ZYTS571)