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
Unité Biologie & Recherche Médicale, CNESTEN, Rabat, Maroc.
Cell Mol Biol (Noisy-le-grand). 2011 Oct 15;57 Suppl:OL1581-91. doi: 10.5772/30025.
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.
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。