Ginsenoside Rh2 stimulates the production of mitochondrial reactive oxygen species and induces apoptosis of cervical cancer cells by inhibiting mitochondrial electron transfer chain complex
Ginsenoside Rh2 stimulates the production of mitochondrial reactive oxygen species and induces apoptosis of cervical cancer cells by inhibiting mitochondrial electron transfer chain complex
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DOI：
10.3892/mmr.2021.12513
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发表时间：
2021-12-01
影响因子：
3.4
通讯作者：
Zhang, Wei
中科院分区：
文献类型：
--
作者：
Liu, Ying;Yu, Shiting;Zhang, Wei
人参皂苷 Rh2 (G-Rh2) 是从人参中提取的单体化合物，具有体外和体内抗癌活性。此前，我们报道G-Rh2诱导HeLa宫颈癌细胞凋亡，该过程与活性氧（ROS）积累和线粒体功能障碍有关。然而，G-Rh2 的上游机制及其细胞靶标仍有待阐明。在本研究中，Cell Counting Kit-8 检测、流式细胞术和 Hoechst 染色显示，G-Rh2 显着抑制细胞活力并诱导宫颈癌细胞凋亡。然而，G-Rh2 被证明对 End1/e6e7 细胞无毒。 JC-1、罗丹明 123 染色、氧化磷酸化和糖酵解能力测定表明，G-Rh2 暴露导致线粒体跨膜电位立即降低，因为它抑制线粒体氧化磷酸化和糖酵解，这两者都会减少细胞 ATP 的产生。蛋白质印迹和电子传递链（ETC）活性测定表明，G-Rh2显着抑制ETC复合物I、III和V的活性。ETC复合物III的过表达部分显着恢复线粒体ROS并抑制G-Rh2诱导的宫颈癌细胞凋亡。分子对接中结合能的预测结果证实，G-Rh2极有可能通过靶向ETC复合物，特别是ETC复合物III来诱导线粒体ROS产生并促进细胞凋亡。总而言之，目前的结果揭示了 G-Rh2 的潜在抗宫颈癌活性，并为 ETC 复合物活性受损导致宫颈癌细胞死亡提供了直接证据。
Ginsenoside Rh2 (G-Rh2) is a monomeric compound that extracted from ginseng and possesses anti-cancer activities both in vitro and in vivo. Previously, we reported that G-Rh2 induces apoptosis in HeLa cervical cancer cells and that the process was related to reactive oxygen species (ROS) accumulation and mitochondrial dysfunction. However, the upstream mechanisms of G-Rh2, along with its cellular targets, remain to be elucidated. In the present study, the Cell Counting Kit-8 assay, flow cytometry and Hoechst staining revealed that G-Rh2 significantly inhibited cell viability and induced apoptosis of cervical cancer cells. However, G-Rh2 was demonstrated to be non-toxic to End1/e6e7 cells. JC-1, rhodamine 123 staining, oxidative phosphorylation and glycolysis capacity assays demonstrated that G-Rh2 exposure caused an immediate decrease in mitochondrial transmembrane potential due to its inhibition of mitochondrial oxidative phosphorylation, as well as glycolysis, both of which reduced cellular ATP production. Western blotting and electron transport chain (ETC) activity assays revealed that G-Rh2 significantly inhibited the activity of ETC complexes I, III and V. Overexpression of ETC complex III partially significantly restored mitochondrial ROS and inhibited the apoptosis of cervical cancer cells induced by G-Rh2. The predicted results of binding energy in molecular docking, confirmed that G-Rh2 was highly likely to induce mitochondrial ROS production and promote cell apoptosis by targeting the ETC complex, especially for ETC complex III. Taken together, the present results revealed the potential anti-cervical cancer activity of G-Rh2 and provide direct evidence for the contribution of impaired ETC complex activity to cervical cancer cell death.