Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran.
Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran; Cell and Molecular Biotechnology Research Group, Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran.
Life Sci. 2014 Jul 30;109(2):87-94. doi: 10.1016/j.lfs.2014.06.006. Epub 2014 Jun 20.
In this research, ferutinin was evaluated for its possible cytotoxic and apoptotic inducing effects in vitro and in vivo.
To determine IC50 values of ferutinin, CT26, HT29 and NIH/3T3 cells were treated with different concentrations of ferutinin. In addition to morphological changes in cells, the DNA damage was studied using DAPI staining, comet assay and PI staining. Ferutinin was also tested for its in vivo activity.
Analyses of cell survival by MTT assay showed that the IC50 values of ferutinin on CT26 and HT29 cells were 26 and 29 μg/ml, respectively, while after treating nontumoural mouse cells even with 50 μg/ml ferutinin, 70% of cells was still surviving. The results of DAPI staining and comet assay revealed that ferutinin significantly induced DNA damage in treated cells. Induction of sub-G1 peak after PI staining was also indicative of apoptotic effects of ferutinin in cancerous cells. In vivo studies showed a significant regression in tumour size in mice treated with ferutinin as compared to control groups. Its antitumour effects were very similar to the cisplatin treated group. Histological studies demonstrated that apoptosis rate in tumour cells was increased in comparison to tumour cells in control mice without ferutinin treatment. Interestingly, haematoxylin and eosin staining showed no damage in the spleen and liver of ferutinin treated mice.
As ferutinin showed less toxic effects in nontumoural cells, and induced its effects via apoptosis induction, it could be considered as an effective anticancer agent for future preclinical experiments.
本研究旨在评估 ferutinin 在体外和体内的潜在细胞毒性和促凋亡作用。
为了确定 ferutinin 的 IC50 值,用不同浓度的 ferutinin 处理 CT26、HT29 和 NIH/3T3 细胞。除了观察细胞形态变化外,还通过 DAPI 染色、彗星试验和 PI 染色研究 DNA 损伤。还测试了 ferutinin 的体内活性。
MTT 法分析细胞存活率的结果表明,ferutinin 对 CT26 和 HT29 细胞的 IC50 值分别为 26 和 29μg/ml,而用 50μg/ml ferutinin 处理非肿瘤性小鼠细胞后,仍有 70%的细胞存活。DAPI 染色和彗星试验的结果表明,ferutinin 能显著诱导处理细胞的 DNA 损伤。PI 染色后出现亚 G1 峰也表明 ferutinin 在癌细胞中具有促凋亡作用。体内研究表明,与对照组相比,用 ferutinin 治疗的小鼠肿瘤体积明显缩小。其抗肿瘤作用与顺铂治疗组非常相似。组织学研究表明,与未用 ferutinin 治疗的对照组肿瘤细胞相比,肿瘤细胞的凋亡率增加。有趣的是,苏木精-伊红染色显示,用 ferutinin 治疗的小鼠的脾和肝没有损伤。
由于 ferutinin 在非肿瘤细胞中毒性较小,并通过诱导细胞凋亡发挥作用,因此它可被视为未来临床前实验的一种有效的抗癌药物。