HSPA1A or β-actin 华东医科大学(英文版).pdfVIP

Cell Stress and Chaperones DOI 10.1007/s12192-012-0374-y ORIGINAL PAPER Development of rapid and highly sensitive HSPA1A promoter-driven luciferase reporter system for assessing oxidative stress associated with low-dose photodynamic therapy Yuanhong Zheng Vanminh Le Zhuoan Cheng Sheng Xie Hegeng Li Jianhui Tian Jianwen Liu Received: 6 August 2012 /Accepted: 7 September 2012 # Cell Stress Society International 2012 Abstract Photodynamic therapy (PDT) is a regulatory- when compared with a cell viability test based on ATP quan- approved modality for treating a variety of malignant tumors. tification and ROS levels. Furthermore, phthalocyanine zinc It induces tumor tissue damage via photosensitizer-mediated and methylene blue both induced significantly elevated levels oxidative cytotoxicity. The heat shock protein 70 (HSP70-1) is of relative luciferase activity in a dose-dependent manner. a stress protein encoded by the HSPA1A gene and is signifi- cantly induced by oxidative stress associated with PDT. The Keywords Photodynamic therapy (PDT) . HSPA1A aim of this study was to identify the functional region of the promoteractivity . Stressoxidative .Luciferasereporter .Cell HSPA1A promoter that responds to PDT-induced oxidative viability stress and uses the stress responsiveness of HSPA1A expres- sion to establish a rapid and cost-effective photocytotoxic Abbreviations assessment bioassay to evaluate the photodynamic potential HSP Heat shock protein of photosensitizers. By constructing luciferase vectors with a PDT Photodynamic therapy variety of hspa1a promoter fractions and examining their ROS Reactive oxygen species relative luciferase activity, we demonstrated that the DNA DMSO Dimethyl sulfoxide sequence from −218 to +87 of the HSPA1A gene could be