人参皂苷Rg5的安全性及其对宫颈癌Hela细胞的体外抑制活性

Study on In Vitro Inhibitory Activity and Safety of Ginsenoside Rg5 against Hela Cells

摘要:

为探讨人参皂苷Rg5对癌细胞的抑制作用和安全性，使用CCK-8法筛选敏感细胞系，后以多种手段评估作用机制和安全性。以流式细胞术检测Rg5对敏感细胞凋亡和周期的影响、以划痕和Transwell实验考察Rg5对细胞迁移和侵袭的抑制、以Western Blot法探究Rg5对细胞特定蛋白的扰动、以Auto Dock模拟Rg5与MMP-2和MMP-9的结合、以体内实验验证Rg5的安全性。结果显示，在Hela、MCF-7、HCT116、HepG2、A549、SiHa、CaSki细胞中，宫颈癌Hela细胞对Rg5最敏感，72 h半抑制浓度为53.02 μmol·L-1。Rg5能诱导Hela细胞凋亡、将其阻滞于G2/M期并抑制迁移和侵袭。Western Blot实验表明，Rg5能扰动周期相关蛋白并触发线粒体凋亡通路，还能降低MMP-2、MMP-9、E-cadherin和干性因子的表达。Auto Dock显示，Rg5能结合在MMP-2和MMP-9的疏水口袋中。安全性实验表明，Rg5对小鼠肝肾功能、血常规指标和主要器官无明显毒性。作为一种天然产物，Rg5能通过触发凋亡通路诱导细胞凋亡，通过下调癌细胞干性因子和相关蛋白的表达抑制Hela细胞的侵袭。研究结果为Rg5在功能性食品、保健品方面的进一步开发利用提供了理论支持。

Abstract:

To study the inhibitory effect, mechanism, and safety of ginsenosides Rg5 towards cancer cells, CCK-8 assay was conducted to screen the sensitive cancer cell line. Then the mechanisms and safety of Rg5 were studied via various of assays. The flow cytometry was applied to evaluate the apoptosis and cell cycle of sensitive cancer cells treated with Rg5. The cancer cell migration and invasion were studied via scratching experiment and transwell assay. Besides, the disturbance of protein expression in sensitive cells treated with Rg5 was measured by Western Blot experiment. The binding modes between Rg5 and MMP-2 or MMP-9 were simulated by Auto Dock study. Furthermore, the safety of Rg5 was evaluated via animal experiments. The results indicated that Hela cells were sensitive towards Rg5 among the Hela, MCF-7, HCT116, HepG2, A549, SiHa, and CaSki cells. The 72 h-IC50 value was 53.02 μmol·L-1. The Rg5 could induce Hela cell apoptosis, arrest Hela cells at G2/M phase, and inhibit migration and invasion of Hela cells. The results of Western Blot assay demonstrated that Rg5 could activate the mitochondrial apoptosis pathway, reduce the expression of MMP-2, MMP-9, E-cadherin, and cancer cell stemness factors. Auto Dock study indicated that Rg5 could bind into the hydrophobic pockets of MMP-2 and MMP-9. The in vivo safety experiments showed that Rg5 exhibited no obvious disturbance or damage towards the function of liver and kidney, the indexes of blood routine, and the major organs of mice. As a natural produc, Rg5 could induce apoptosis via initiating the apoptosis pathway and suppress cell invasion via down-regulating the expression of cancer stemness related factors, MMP-2, and MMP-9. The results provided theoretical support for the further development and utilization of Rg5 in the field of functional food and health products.