Investigating Apoptotic Effects of Methanolic Extract of Dorema glabrum Seed on WEHI-164 Cells.

ISRN Pharmacology Pub Date : 2013-07-17 eCollection Date: 2013-01-01 DOI:10.1155/2013/949871
Maryam Bannazadeh Amirkhiz, Nadereh Rashtchizadeh, Hossein Nazemiyeh, Jalal Abdolalizadeh, Leila Mohammadnejad, Behzad Baradaran
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引用次数: 13

Abstract

We aimed to investigate the apoptotic effects of the methanolic extract of Dorema glabrum seed on WEHI-164, cancerous cells in comparison with L929, normal cells and compared them with the cytotoxic effects of Taxol. So, MTT test and DNA fragmentation assay were performed on cultured and treated cells. Also electrophoresis which was followed by immunoblotting was done to survey the production of Caspase-3 and Bcl2 proteins, and to inquire into their relative genes expression, RT-PCR was used. According to our findings, the methanolic extract of Dorema glabrum seed can alter cells morphology as they shrink and take a spherical shape and lose their attachment too. So, the plant extract inhibits cell growth albeit in a time- and dose-dependent manner and results in degradation of chromosomal DNA. Induction of apoptosis by the plant extract was proved by the reduction of pro-Caspase-3 and Bcl2 proteins and increase in Caspase-3 gene expression and decrease in that of bcl2 too. Our data well established the antiproliferative effect of methanolic extract of Dorema glabrum seed and clearly showed that the plant extract can induce apoptosis and not necrosis in vitro. These results demonstrated that Dorema glabrum seed might be a novel and attractive therapeutic candidate for tumor treatment.

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梦莲种子甲醇提取物对WEHI-164细胞凋亡作用的研究。
本实验目的是通过与L929、正常细胞进行比较,探讨多丽麻种子甲醇提取物对WEHI-164、癌细胞的凋亡作用,并与紫杉醇的细胞毒作用进行比较。因此,对培养细胞和处理细胞进行MTT试验和DNA片段化试验。采用电泳和免疫印迹法检测Caspase-3和Bcl2蛋白的产生情况,并采用RT-PCR法研究其相关基因的表达情况。根据我们的研究结果,杜鹃种子的甲醇提取物可以改变细胞的形态,使细胞收缩,呈球形,并失去附着力。因此,植物提取物抑制细胞生长,尽管以时间和剂量依赖的方式,并导致染色体DNA的降解。通过降低Caspase-3和Bcl2蛋白,增加Caspase-3基因表达,降低Bcl2基因表达,证实了植物提取物诱导细胞凋亡的作用。我们的实验结果很好地证实了梦麻种子甲醇提取物的抗增殖作用,并清楚地表明该植物提取物在体外诱导细胞凋亡而非坏死。这些结果表明,脑光豆种子可能是一种新的有吸引力的肿瘤治疗候选药物。
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