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金缕梅正己烷萃取物对人宫颈癌(HeLa)细胞的抗增殖和促凋亡作用。

Antiproliferative and apoptogenic effects of Cassia fistula L. n-hexane fraction against human cervical cancer (HeLa) cells.

机构信息

Department of Botanical and Environmental Sciences, Guru Nanak Dev University, Amritsar, Punjab, 143005, India.

Post Graduate Department of Botany, Khalsa College, Amritsar, India.

出版信息

Environ Sci Pollut Res Int. 2020 Sep;27(25):32017-32033. doi: 10.1007/s11356-020-08916-9. Epub 2020 Jun 6.

Abstract

The current study was performed to evaluate the antiproliferative and apoptosis-inducing potential of n-hexane fraction from Cassia fistula L. (Caesalpinioideae) fruits. The antiproliferative property of the fraction was determined by MTT assay against cancer cell lines including HeLa, MG-63, IMR-32, and PC-3 with GI value of 97.69, 155.2, 143, and 160.2 μg/ml respectively. The fraction was further explored for its apoptotic effect using confocal, SEM, and flow cytometry studies in HeLa cells. It was observed that the treatment of fraction revealed fragmentation of DNA, chromatin condensation, membrane blebbing, and formation of apoptotic bodies in a dose-dependent manner. The fraction also showed a remarkable increase in the level of ROS, mitochondrial depolarization and G/G phase cell cycle arrest, and induction in the phosphatidylserine externalization analyzed using Annexin V-FITC/PI double staining assay in HeLa cells. Kaempferol, Ellagic acid, and Epicatechin are the major phytoconstituents present in the fraction as revealed by the HPLC. The treatment of n-hexane fraction showed downregulation in the gene expression of Bcl-2 and upregulation in the expression level of p53, Bad, and caspase-3 genes analyzed using semi-quantitative RT-PCR in HeLa cells. These results suggest that n-hexane fraction from C. fistula inhibited the proliferation of cervical cancer cells efficiently by the induction of apoptosis. Graphical abstract.

摘要

本研究旨在评估苏木(豆科)果实正己烷部分的抗增殖和诱导细胞凋亡作用。通过 MTT 法测定该部分对包括 HeLa、MG-63、IMR-32 和 PC-3 在内的癌细胞系的增殖抑制作用,GI 值分别为 97.69、155.2、143 和 160.2μg/ml。进一步通过共聚焦、SEM 和流式细胞术研究观察该部分在 HeLa 细胞中的凋亡作用。结果表明,该部分以剂量依赖的方式导致 DNA 片段化、染色质浓缩、细胞膜起泡和凋亡小体形成。该部分还显示出 ROS 水平、线粒体去极化和 G1/G0 细胞周期停滞的显著增加,并通过 Annexin V-FITC/PI 双重染色分析在 HeLa 细胞中诱导磷脂酰丝氨酸外翻。HPLC 分析表明,该部分含有主要的植物成分,如山奈酚、鞣花酸和表儿茶素。正己烷部分处理后,通过半定量 RT-PCR 分析在 HeLa 细胞中观察到 Bcl-2 基因表达下调和 p53、Bad 和 caspase-3 基因表达上调。这些结果表明,苏木正己烷部分通过诱导细胞凋亡有效抑制宫颈癌细胞的增殖。

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