体外亚甲蓝对骨肉瘤细胞的光动力作用。

Photodynamic action of methylene blue in osteosarcoma cells in vitro.

机构信息

School of Chinese Medicine, The Chinese University of Hong Kong, Shatin, Hong Kong; School of Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China.

School of Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China.

出版信息

Photodiagnosis Photodyn Ther. 2014 Mar;11(1):13-9. doi: 10.1016/j.pdpdt.2013.09.003. Epub 2013 Oct 16.

DOI:10.1016/j.pdpdt.2013.09.003

PMID:24629696

Abstract

BACKGROUND

Osteosarcoma is a common malignant bone tumor which threatens the life of young people worldwide. To explore alternative strategy for combating osteosarcoma, a light-emitting diode (LED) that activates methylene blue (MB) was used in the present study to investigate cell death of osteosarcoma-derived UMR106 cells.

MATERIALS AND METHODS

Photocytotoxicity in UMR106 cells was investigated 24h after photodynamic activation of MB using sulforhodamine B (SRB) assay and light microscopy. Apoptosis induction was observed 24h after photodynamic treatment using a confocal laser scanning microscopy (CLSM) with Hoechst 33342 staining. The change in mitochondrial membrane potential (MMP) was analyzed using a flow cytometry with rhodamine 123 staining.

RESULTS

MB under red light irradiation caused a drug-concentration (0-100μM) and light-dose (0-32J/cm(2)) dependent cytotoxicity in UMR106 cells. The SRB assay and light microscopy observed a significant decrease in the number of UMR106 cells attached to the bottom of culture well after LED light-activated MB (100μM, 32J/cm(2)). Nuclear shrinkage, chromatin condensation and fragmentation were found in the treated cells by nuclear staining. In addition, flow cytometry showed that the MMP in UMR106 cells was rapidly reduced by photo-activated MB (100μM, 32J/cm(2)).

CONCLUSION

Photodynamic action of MB under LED irradiation could remarkably kill osteosarcoma cells and induce cell apoptosis as well as MMP collapse.

摘要

背景

骨肉瘤是一种常见的恶性骨肿瘤，它威胁着全世界年轻人的生命。为了探索对抗骨肉瘤的替代策略，本研究使用发光二极管（LED）激活亚甲蓝（MB）来研究骨肉瘤衍生的 UMR106 细胞的细胞死亡。

材料与方法

使用磺基罗丹明 B（SRB）测定法和光镜观察 MB 光动力激活 24 小时后 UMR106 细胞的光细胞毒性。用 Hoechst 33342 染色的共聚焦激光扫描显微镜（CLSM）观察光动力处理 24 小时后细胞凋亡的诱导。用 rhodamine 123 染色的流式细胞术分析线粒体膜电位（MMP）的变化。

结果

MB 在红光照射下引起 UMR106 细胞的药物浓度（0-100μM）和光剂量（0-32J/cm²）依赖性细胞毒性。SRB 测定法和光镜观察到 LED 激活 MB（100μM，32J/cm²）后贴附在培养皿底部的 UMR106 细胞数量明显减少。细胞核染色显示处理后的细胞出现核收缩、染色质浓缩和碎裂。此外，流式细胞术显示光激活 MB（100μM，32J/cm²）可迅速降低 UMR106 细胞的 MMP。