果实提取物及其生物活性部位通过上调 p53/γH2AX 信号和 MIA PaCa-2 细胞 G2/M 期阻滞抑制增殖作用。
Antiproliferative Effects of Fruit Extract and its Bioactive Fraction through Upregulation of p53/γH2AX Signals and G2/M Phase Arrest in MIA PaCa-2 Cells.
机构信息
Department of Post Graduate Studies and Research in Biotechnology, Kuvempu University, Shankaraghatta 577451, Karnataka, India.
Research Unit of DNA Barcoding of Thai Medicinal Plants, Department of Pharmacognosy and Pharmaceutical Botany, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand.
出版信息
Anticancer Agents Med Chem. 2022;22(17):2998-3008. doi: 10.2174/1871520622666220201103431.
BACKGROUND
Artabotrys odoratissimus (Annonaceae) is a medicinal and ornamental plant widely cultivated in Southeast Asia for its famous ylang ylang essential oil. The fruits of this plant are used for health benefits, but very little is studied about the bioactive principles, their role in regulating oxidative stress and tumour progression.
OBJECTIVE
The study aimed to evaluate the antiproliferative effects of fruit extract of Artabotrys odoratissimus and its bioactive fraction using cell-based assays.
METHODS
The free radical scavenging and antiproliferative effects of Artabotrys odoratissimus fruit ethyl acetate (FEA) extract and its bioactive fraction were evaluated using cell viability assays, colony formation assay, double staining assay, reactive oxygen species (ROS) assay, comet assay, cell cycle analysis, and western blotting.
RESULTS
The extract showed phenolic content of 149.8±0.11μg/mg Gallic acid equivalents and flavonoid content of 214.47±4.18 μg/mg Quercetin. FEA showed an IC value of 76.35 μg/ml in the ABTS assay and an IC value of 134.3±7.8 μg/ml on MIA PaCa-2 cells. The cells treated with 125 μg/ml and 250 μg/ml FEA showed increased apoptotic cells in Double staining assay, DNA damage during comet assay, enhanced ROS, and cell cycle arrest at G2M phase at 125 μg/ml and 250 μg/ml. The active fraction AF5 showed an IC value of 67±1.26 μg/ml on MIA PaCa-2 cells during MTT assay, displayed potential antiproliferative effects, and showed a marked increase in the expression of γH2AX and p53.
CONCLUSION
These results prove that the fruit extract and the bioactive fraction demonstrate oxidative stress-mediated DNA damage, leading to apoptosis in the MIA PaCa-2 cell line.
背景
Artabotrys odoratissimus（番荔枝科）是一种药用和观赏植物，因其著名的依兰依兰精油而在东南亚广泛种植。这种植物的果实被用于保健，但对其生物活性成分、它们在调节氧化应激和肿瘤进展中的作用的研究却很少。
目的
本研究旨在评估 Artabotrys odoratissimus 果实提取物及其生物活性部分对细胞的抗增殖作用。
方法
采用细胞活力测定法、集落形成试验、双染试验、活性氧（ROS）测定法、彗星试验、细胞周期分析和 Western blot 法评价 Artabotrys odoratissimus 果实乙酸乙酯（FEA）提取物及其生物活性部分的自由基清除和抗增殖作用。
结果
提取物的总酚含量为 149.8±0.11μg/mg 没食子酸当量，总黄酮含量为 214.47±4.18μg/mg 槲皮素。ABTS 法测定 FEA 的 IC 值为 76.35μg/ml，MIA PaCa-2 细胞的 IC 值为 134.3±7.8μg/ml。用 125μg/ml 和 250μg/ml 的 FEA 处理的细胞，在双染试验中显示出凋亡细胞增加，彗星试验中显示出 DNA 损伤，ROS 增强，以及在 125μg/ml 和 250μg/ml 时细胞周期停滞在 G2M 期。活性部分 AF5 在 MTT 试验中对 MIA PaCa-2 细胞的 IC 值为 67±1.26μg/ml，显示出潜在的抗增殖作用，并显著增加了 γH2AX 和 p53 的表达。
结论
这些结果证明，果实提取物及其生物活性部分在 MIA PaCa-2 细胞系中显示出氧化应激介导的 DNA 损伤，导致细胞凋亡。
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