秘鲁枸杞（黄金莓）中的 4β-羟基乌苏烷内酯 E 通过 DNA 损伤、细胞凋亡和 G2/M 期阻滞抑制人肺癌细胞的生长。

BACKGROUND

The crude extract of the fruit bearing plant, Physalis peruviana (golden berry), demonstrated anti-hepatoma and anti-inflammatory activities. However, the cellular mechanism involved in this process is still unknown.

METHODS

Herein, we isolated the main pure compound, 4beta-Hydroxywithanolide (4betaHWE) derived from golden berries, and investigated its antiproliferative effect on a human lung cancer cell line (H1299) using survival, cell cycle, and apoptosis analyses. An alkaline comet-nuclear extract (NE) assay was used to evaluate the DNA damage due to the drug.

RESULTS

It was shown that DNA damage was significantly induced by 1, 5, and 10 microg/mL 4betaHWE for 2 h in a dose-dependent manner (p < 0.005). A trypan blue exclusion assay showed that the proliferation of cells was inhibited by 4betaHWE in both dose- and time-dependent manners (p < 0.05 and 0.001 for 24 and 48 h, respectively). The half maximal inhibitory concentrations (IC50) of 4betaHWE in H1299 cells for 24 and 48 h were 0.6 and 0.71 microg/mL, respectively, suggesting it could be a potential therapeutic agent against lung cancer. In a flow cytometric analysis, 4betaHWE produced cell cycle perturbation in the form of sub-G1 accumulation and slight arrest at the G2/M phase with 1 microg/mL for 12 and 24 h, respectively. Using flow cytometric and annexin V/propidium iodide immunofluorescence double-staining techniques, these phenomena were proven to be apoptosis and complete G2/M arrest for H1299 cells treated with 5 microg/mL for 24 h.

CONCLUSIONS

In this study, we demonstrated that golden berry-derived 4betaHWE is a potential DNA-damaging and chemotherapeutic agent against lung cancer.

背景

水果植物酸浆（灯笼果）的粗提物具有抗肝癌和抗炎活性。然而，这一过程中涉及的细胞机制尚不清楚。

方法

本文从灯笼果中分离出主要的纯化合物 4β-羟基茄尼醇（4βHWE），并通过生存、细胞周期和凋亡分析研究其对人肺癌细胞系（H1299）的抗增殖作用。碱性彗星核提取物（NE）试验用于评估药物引起的 DNA 损伤。

结果

结果表明，DNA 损伤在剂量依赖性方式下，由 1、5 和 10μg/ml 的 4βHWE 在 2 小时内显著诱导（p<0.005）。台盼蓝排除试验显示，4βHWE 以剂量和时间依赖性方式抑制细胞增殖（24 和 48 小时分别为 p<0.05 和 0.001）。4βHWE 在 H1299 细胞中 24 和 48 小时的半最大抑制浓度（IC50）分别为 0.6 和 0.71μg/ml，表明其可能是一种治疗肺癌的潜在药物。在流式细胞术分析中，4βHWE 以亚 G1 积累的形式产生细胞周期扰动，并在 1μg/ml 时分别在 G2/M 期有轻微阻滞 12 和 24 小时。使用流式细胞术和 Annexin V/碘化丙啶免疫荧光双重染色技术，这些现象被证明是用 5μg/ml 处理 24 小时的 H1299 细胞凋亡和完全 G2/M 阻滞。