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委陵菜乙酸乙酯提取物对骨肉瘤癌细胞的体外抗肿瘤活性。

In vitro antitumor activity of the ethyl acetate extract of Potentilla chinensis in osteosarcoma cancer cells.

作者信息

Wan Guang, Tao Jin-Gang, Wang Guo-Dong, Liu Shen-Peng, Zhao Hong-Xing, Liang Qiu-Dong

机构信息

Department of Orthopedics, The First Affiliated Hospital of Xinxiang Medical University, Weihui, Henan 453100, P.R. China.

出版信息

Mol Med Rep. 2016 Oct;14(4):3634-40. doi: 10.3892/mmr.2016.5679. Epub 2016 Aug 26.

Abstract

The aim of the current study was to evaluate the anticancer effect of the ethanol extract of Potentilla chinensis, a Chinese medicinal plant. An MTT assay was used to evaluate the cell viability of MG‑63 human osteosarcoma cancer cells and fR‑2 cells. Furthermore, the effect of the extract on apoptosis induction, cell cycle phase distribution and inhibition of cell migration in the MG63 human osteosarcoma cancer cell line was evaluated. The effect of the extract on cell cycle phase distribution was assessed by flow cytometry using propidium iodide (PI). Phase contrast microscopy detected the morphological changes in MG63 cancer cells following extract treatment. The results of the study demonstrated that the extract was cytotoxic to MG63 cancer cells, while the normal cell line (epithelial cell line) showed lower susceptibility. Phase contrast microscopy showed distinguishing morphological features, such as cell shrinkage and blebbing induced by the extract treatment in osteosarcoma cancer cells. The average proportion of Annexin V‑positive cells (total apoptotic cells) significantly increased from 5.6% in the control to 24.2, 38.8 and 55.7% in the 40, 80 and 150 µg/ml groups, respectively. The extract induced early and late apoptosis in the cancer cells. Flow cytometric analysis revealed that the extract induced G0/G1‑cell cycle arrest, which also showed significant dose‑dependence. The extract induced a significant and concentration‑dependent reduction in cell migration. Moreover, DNA fragmentation was also examined by observation of the formation of DNA ladders. It was demonstrated that DNA fragmentation was increased with extract concentration compared with that in the control. Taken together, EEPC may serve as potential therapeutic agent against osteosarcoma, provided that the toxicity profile and in vivo investigations demonstrate that it is safe.

摘要

本研究旨在评估中药委陵菜乙醇提取物的抗癌作用。采用MTT法评估MG-63人骨肉瘤癌细胞和fR-2细胞的细胞活力。此外,还评估了该提取物对MG63人骨肉瘤癌细胞系凋亡诱导、细胞周期阶段分布及细胞迁移抑制的影响。使用碘化丙啶(PI)通过流式细胞术评估提取物对细胞周期阶段分布的影响。相差显微镜检测提取物处理后MG63癌细胞的形态变化。研究结果表明,该提取物对MG63癌细胞具有细胞毒性,而正常细胞系(上皮细胞系)的敏感性较低。相差显微镜显示出明显的形态特征,如提取物处理骨肉瘤癌细胞后诱导的细胞收缩和起泡。膜联蛋白V阳性细胞(总凋亡细胞)的平均比例从对照组的5.6%分别显著增加到40、80和150μg/ml组的24.2%、38.8%和55.7%。该提取物诱导癌细胞早期和晚期凋亡。流式细胞术分析显示,该提取物诱导G0/G1期细胞周期阻滞,也表现出显著的剂量依赖性。该提取物诱导细胞迁移显著且呈浓度依赖性降低。此外,还通过观察DNA梯带的形成来检测DNA片段化。结果表明,与对照组相比,DNA片段化随提取物浓度增加而增加。综上所述,委陵菜乙醇提取物可能作为一种潜在的抗骨肉瘤治疗药物,前提是其毒性特征和体内研究表明它是安全的。

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