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刺蒺藜提取物对人宫颈癌细胞的体外抗增殖作用及凋亡诱导作用

In vitro antiproliferative effect and induction of apoptosis by Retama monosperma L. extract in human cervical cancer cells.

作者信息

Merghoub N, Benbacer L, El Btaouri H, Ait Benhassou H, Terryn C, Attaleb M, Madoulet C, Benjouad A, El Mzibri M, Morjani H, Amzazi S

机构信息

Unité Biologie & Recherche Médicale, CNESTEN, Rabat, Maroc.

出版信息

Cell Mol Biol (Noisy-le-grand). 2011 Oct 15;57 Suppl:OL1581-91. doi: 10.5772/30025.

Abstract

The antiproliferative effect of different extracts obtained from Retama monosperma L. was investigated on human SiHa and HeLa cervical cancer cell lines using a MTT colorimetric assay. The Retama monosperma L. dichloromethane fraction (Rm-DF) was the most active extract, exhibiting a significant cytotoxic activity on both cell lines in a dose-dependent manner, after 72 h of treatment. IC50 values obtained were 14.57 ± 4.15 μg/ml and 21.33 ± 7.88 μg/ml, for SiHa and HeLa cell lines respectively. The morphological features assessment of apoptosis in Rm-DF-treated cells showed a condensation of chromatin and apoptotic bodies, accompanied by a decrease in mitochondrial membrane potential (ΔΨm) and an increase in reactive oxygen species in both cell lines. The induction of apoptosis was further confirmed by Western blotting pro-caspase 3, Bcl2 and PARP; caspase 3 activity assay; and Annexin V labelling. Analysis of Rm-DF by CG/MS revealed the presence of five known quinolizidine alkaloids as well as, sparteine (10,97%), L-methyl cytisine (9.11%), 17-oxosparteine (3.49%), lupanine (0.93%) and anagyrine (39.63%). This study shows that Retama monosperma L. extract exhibits a potential anticancer activity against cervical cancer cell lines in vitro through the inhibition of proliferation and induction of apoptosis, which may involve a mitochondria-mediated signaling pathway.

摘要

采用MTT比色法研究了单籽骆驼刺(Retama monosperma L.)不同提取物对人SiHa和HeLa宫颈癌细胞系的抗增殖作用。单籽骆驼刺二氯甲烷提取物(Rm-DF)是活性最强的提取物,在处理72小时后,对两种细胞系均表现出显著的剂量依赖性细胞毒性活性。SiHa和HeLa细胞系的半数抑制浓度(IC50)值分别为14.57±4.15μg/ml和21.33±7.88μg/ml。对Rm-DF处理的细胞进行凋亡形态学特征评估,结果显示两种细胞系均出现染色质凝聚和凋亡小体,同时线粒体膜电位(ΔΨm)降低,活性氧增加。通过蛋白质免疫印迹法检测前半胱天冬酶-3、Bcl-2和聚(ADP-核糖)聚合酶(PARP)、半胱天冬酶-3活性测定以及膜联蛋白V标记,进一步证实了细胞凋亡的诱导。通过气相色谱/质谱联用(CG/MS)分析Rm-DF,结果显示存在5种已知的喹诺里西啶生物碱,即鹰爪豆碱(10.97%)、L-甲基金雀花碱(9.11%)、17-氧代鹰爪豆碱(3.49%)、羽扇豆碱(0.93%)和臭豆碱(39.63%)。本研究表明,单籽骆驼刺提取物通过抑制增殖和诱导凋亡,在体外对宫颈癌细胞系具有潜在的抗癌活性,这可能涉及线粒体介导的信号通路。

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