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萤火虫荧光素酶生物发光介导的癌细胞光动力毒性评估。

Evaluation of firefly luciferase bioluminescence mediated photodynamic toxicity in cancer cells.

作者信息

Schipper Meike L, Patel Manishkumar R, Gambhir Sanjiv S

机构信息

Molecular Imaging Program at Stanford (MIPS), Department of Radiology and Bio-X Program, Stanford University, E 150 Clark Center, 318 Campus Drive, Palo Alto, CA 94305-5427, USA.

出版信息

Mol Imaging Biol. 2006 Jul-Aug;8(4):218-25. doi: 10.1007/s11307-006-0048-1.

Abstract

PURPOSE

This work investigated whether fLuc-catalyzed oxidation of D-luciferin generates sufficient light to induce photodynamic toxicity in cancer cells.

PROCEDURES

Light emission was assessed via cooled CCD (charge-coupled device) camera. Parental and fLuc expressing cancer cells were exposed to subtoxic concentrations of photosensitizers (Rose Bengal or hypericin) and D-luciferin, sunlight, or lamplight. Toxicity was assessed by MTT assay.

RESULTS

fLuc expressing cells emitted up to 500-fold higher levels of photons than parental cell lines. Although exposure to photosensitizer and sunlight reduced survival of various cell lines, survival of fLuc expressing cells incubated with photosensitizer and D-luciferin, or photosensitizer and lamplight, did not differ significantly from parental or untreated cells.

CONCLUSIONS

Contesting recent reports, fLuc bioluminescence does not generate sufficient photons to induce Rose Bengal or hypericin photodynamic toxicity in a range of malignant and nonmalignant cell lines, and is not suitable as a generalizable approach to antineoplastic therapy.

摘要

目的

本研究旨在探讨荧光素酶(fLuc)催化氧化D - 荧光素所产生的光是否足以诱导癌细胞发生光动力毒性。

程序

通过冷却电荷耦合器件(CCD)相机评估发光情况。将亲本癌细胞和表达fLuc的癌细胞暴露于亚毒性浓度的光敏剂(孟加拉玫瑰红或金丝桃素)、D - 荧光素、阳光或灯光下。通过MTT法评估毒性。

结果

表达fLuc的细胞发出的光子水平比亲本细胞系高出多达500倍。虽然暴露于光敏剂和阳光会降低各种细胞系的存活率,但与光敏剂和D - 荧光素或光敏剂和灯光一起孵育的表达fLuc的细胞的存活率与亲本细胞或未处理细胞相比没有显著差异。

结论

与最近的报道相反,fLuc生物发光产生的光子不足以在一系列恶性和非恶性细胞系中诱导孟加拉玫瑰红或金丝桃素光动力毒性,因此不适合作为一种通用的抗肿瘤治疗方法。

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