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毛竹提取物诱导骨肉瘤细胞凋亡信号,与AMPK激活有关。

Phyllostachys edulis extract induces apoptosis signaling in osteosarcoma cells, associated with AMPK activation.

作者信息

Chou Chi-Wen, Cheng Ya-Wen, Tsai Chung-Hung

机构信息

Institute of Medicine, Chung Shan Medical University, Taichung, Taiwan ; Department of Orthopedics Surgery, National Taiwan University Hospital Yun-Lin Branch, Yun-Lin, Taiwan.

Graduate Institute of Cancer Biology and Drug Discovery, College of Medical Science and Technology, Taipei Medical University, Taipei, Taiwan.

出版信息

Drug Des Devel Ther. 2014 Sep 24;8:1577-84. doi: 10.2147/DDDT.S69342. eCollection 2014.

Abstract

OBJECTIVE

Bamboo is distributed worldwide, and its different parts are used as foods or as a traditional herb. Recently, antitumoral effects of bamboo extracts on several tumors have been increasingly reported; however, antitumoral activity of bamboo extracts on osteosarcoma remains unclear. In the present study, we investigated effects of an aqueous Phyllostachys edulis leaf extract (PEE) on osteosarcoma cells and the underlying mechanism of inhibition.

METHODS

The growth of human osteosarcoma cell lines 143 B and MG-63 and lung fibroblast MRC-5 cells was determined by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Apoptosis was demonstrated using TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling) assay and flow cytometric analysis. Phosphorylation and protein levels were determined by immunoblotting.

RESULTS

After treatment with PEE, viability of 143 B and MG-63 cells was dose-dependently reduced to 36.3% ± 1.6% of control values, which were similar to AICAR (5-aminoimidazole-4-carboxamide 1-β-D-ribofuranoside) treatments. In parallel, ratios of apoptotic cells and cells in the sub-G1 phase were significantly increased. Further investigation showed that PEE treatments led to activation of caspase cascades and changes of apoptotic mediators Bcl2, Bax, and p53. Consistently, our results revealed that PEE activated adenosine monophosphate-activated protein kinase (AMPK) signaling, and the AMPK activation was associated with the induction of apoptotic signaling.

CONCLUSION

Our results indicated that PEE suppressed the growth of 143 B and MG-63 cells but moderately affected MRC-5 cells. PEE-induced apoptosis may attribute to AMPK activation and the following activation of apoptotic signaling cascades. These findings revealed that PEE possesses antitumoral activity on human osteosarcoma cells by manipulating AMPK signaling, suggesting that PEE alone or combined with regular antitumor drugs may be beneficial as osteosarcoma treatments.

摘要

目的

竹子分布于世界各地,其不同部位被用作食物或传统草药。最近,越来越多的报道称竹子提取物对多种肿瘤具有抗肿瘤作用;然而,竹子提取物对骨肉瘤的抗肿瘤活性仍不清楚。在本研究中,我们研究了毛竹叶水提取物(PEE)对骨肉瘤细胞的影响及其抑制的潜在机制。

方法

通过MTT(3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐)试验测定人骨肉瘤细胞系143 B和MG-63以及肺成纤维细胞MRC-5细胞的生长。使用TUNEL(末端脱氧核苷酸转移酶dUTP缺口末端标记)试验和流式细胞术分析来证明细胞凋亡。通过免疫印迹法测定磷酸化和蛋白质水平。

结果

用PEE处理后,143 B和MG-63细胞的活力呈剂量依赖性降低至对照值的36.3%±1.6%,这与AICAR(5-氨基咪唑-4-甲酰胺1-β-D-呋喃核糖苷)处理相似。同时,凋亡细胞和亚G1期细胞的比例显著增加。进一步研究表明,PEE处理导致半胱天冬酶级联反应的激活以及凋亡介质Bcl2、Bax和p53的变化。一致地,我们的结果显示PEE激活了腺苷单磷酸激活的蛋白激酶(AMPK)信号通路,并且AMPK的激活与凋亡信号的诱导有关。

结论

我们的结果表明,PEE抑制143 B和MG-63细胞的生长,但对MRC-5细胞有中度影响。PEE诱导的细胞凋亡可能归因于AMPK的激活以及随后凋亡信号级联反应的激活。这些发现表明,PEE通过操纵AMPK信号通路对人骨肉瘤细胞具有抗肿瘤活性,这表明PEE单独使用或与常规抗肿瘤药物联合使用可能对骨肉瘤治疗有益。

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