Induction of apoptosis in tumor cells by three naphthoquinone esters isolated from Thai medicinal plant:: Rhinacanthus nasutus Kurz.
Induction of apoptosis in tumor cells by three naphthoquinone esters isolated from Thai medicinal plant:: Rhinacanthus nasutus Kurz.
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DOI：
10.1248/bpb.29.2070
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发表时间：
2006-10-01
影响因子：
2
通讯作者：
Oku, Naoto
中科院分区：
文献类型：
--
作者：
Siripong, Pongpun;Yahuafai, Jantana;Oku, Naoto
鼻棘（菊科）已被用作泰国传统药物用于治疗各种癌症。最近，我们报道了该植物的活性成分--本文研究了三种主要的萘醌酯类化合物-长叶树通过TUNEL染色、DNA片段化分析、流式细胞术和Asp-Glu-Val-Asp-peptide-nitroanilide（一种caspase-3底物）的切割分析，观察了人宫颈癌（HeLaS 3）细胞中caspase-3的表达。在不同浓度的药物作用于HeLaS 3细胞后，刺五加皂苷-C、-N和-Q对HeLaS 3细胞表现出抗增殖作用，24、48和72 h时间点的IC_（50）值分别为80、659、73 μ M; 55、45、55 μ M; 1.5、1.5和5.0 μ M。48 It暴露后，清楚地观察到显示棘皮素处理的细胞的核碎裂的形态学变化。与该观察结果一致，提取的DNA的琼脂糖凝胶电泳也明显显示梯状结构的出现。流式细胞仪分析显示，在24 h孵育后，海棘毒素-N引起HeLaS 3细胞G2/M期阻滞，并在48和72 h孵育后增加HeLaS 3细胞群中亚G1亚二倍体细胞（凋亡细胞）的比例。此外，药物治疗显著提高了caspase-3的活性。在此基础上，我们的研究结果首次证明了从红毛菊根中分离的三种主要萘醌酯类化合物的抑制作用。长叶树对HeLaS 3细胞生长的抑制作用可能是通过诱导细胞凋亡而产生的，而凋亡可能与caspase-3通路的激活有关。
Rhinacanthus nasutus KURZ. (Acanthaceae) has been used as Thai traditional medicine for the treatment of various cancers. Recently, we reported that rhinacanthins, active components of the plant, had antiproliferative activity against human cancer line cells. In the present study, we investigated the growth inhibitory mechanism of rhinacanthins-C, -N and -Q, three main naphthoquinone esters isolated from the roots of R. nasutus KURZ. in human cervical carcinoma (HeLaS3) cells by means of TUNEL staining, DNA fragmentation assay, flow cytometry, and cleavage assay of Asp-Glu-Val-Asp-peptide-nitroanilide, a caspase-3 substrate. After the HeLaS3 cells was exposed with different concentrations of the drugs, rhinacanthins-C, -N and -Q exhibited antiproliferative effects on HeLaS3 cells with the IC50 values of 80, 659 73 mu M; 55, 45, 55 mu M; and 1.5, 1.5 and 5.0 mu M for 24, 48 and 72 h time points, respectively. Morphological changes showing nuclear fragmentation of rhinacanthins-treated cells were clearly observed after 48 It exposure. Consistent with this observation, the appearance of a ladder formation was also evident with an agarose gel electrophoresis of the extracted DNA. Flow cytometric analysis revealed that rhinacanthin-N caused G2/M arrest of HeLaS3 cells after 24 h incubation, and increased the proportion of sub-G1 hypodiploid cells, apoptotic cells, in the population of HeLaS3 cells after 48 and 72 h incubation. Moreover, the drug treatment markedly elevated the activity of caspase-3. Based on these results, our findings demonstrated for the first time that the inhibitory effects of three main naphthoquinone esters isolated from the roots of R. nasutus KURZ. on the growth of HeLaS3 cells appear to arise from the induction of apoptosis, that might be associated with the activation of caspase-3 pathway.