naa a22 4500 510746454 CHVBK 20180411083048.0 cr unu---uuuuu 180411e20130301xx s 000 0 eng 10.1007/s12192-012-0374-y doi (NATIONALLICENCE)springer-10.1007/s12192-012-0374-y Development of rapid and highly sensitive HSPA1A promoter-driven luciferase reporter system for assessing oxidative stress associated with low-dose photodynamic therapy [Elektronische Daten] [Yuanhong Zheng, Vanminh Le, Zhuoan Cheng, Sheng Xie, Hegeng Li, Jianhui Tian, Jianwen Liu] Photodynamic therapy (PDT) is a regulatory-approved modality for treating a variety of malignant tumors. It induces tumor tissue damage via photosensitizer-mediated oxidative cytotoxicity. The heat shock protein 70 (HSP70-1) is a stress protein encoded by the HSPA1A gene and is significantly induced by oxidative stress associated with PDT. The aim of this study was to identify the functional region of the HSPA1A promoter that responds to PDT-induced oxidative stress and uses the stress responsiveness of HSPA1A expression to establish a rapid and cost-effective photocytotoxic assessment bioassay to evaluate the photodynamic potential of photosensitizers. By constructing luciferase vectors with a variety of hspa1a promoter fractions and examining their relative luciferase activity, we demonstrated that the DNA sequence from −218 to +87 of the HSPA1A gene could be used as a functional promoter to detect the PDT-induced oxidative stress. The maximal relative luciferase activity level of HSPA1A (HSP70-1) induced by hypericin-PDT was nearly nine times that of the control. Our results suggest that the novel reporter gene assay using a functional region of the HSP70A1A promoter has significant advantages for the detection of photoactivity in terms of both speed and sensitivity, when compared with a cell viability test based on ATP quantification and ROS levels. Furthermore, phthalocyanine zinc and methylene blue both induced significantly elevated levels of relative luciferase activity in a dose-dependent manner. Cell Stress Society International, 2012 Photodynamic therapy (PDT) nationallicence HSPA1A promoter activity nationallicence Stress oxidative nationallicence Luciferase reporter nationallicence Cell viability nationallicence HSP : Heat shock protein nationallicence PDT : Photodynamic therapy nationallicence ROS : Reactive oxygen species nationallicence DMSO : Dimethyl sulfoxide nationallicence Hyp : Hypericin nationallicence Me : Methylene blue nationallicence ZnPc : Zinc phthalocyanine nationallicence μM : Micromoles per liter nationallicence MTT : 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide nationallicence Zheng Yuanhong State Key Laboratory of Bioreactor Engineering and Shanghai Key Laboratory of Chemical Biology, School of Pharmacy, East China University of Science and Technology, #268, 130 Meilong Road, 200237, Shanghai, People's Republic of China aut Le Vanminh State Key Laboratory of Bioreactor Engineering and Shanghai Key Laboratory of Chemical Biology, School of Pharmacy, East China University of Science and Technology, #268, 130 Meilong Road, 200237, Shanghai, People's Republic of China aut Cheng Zhuoan State Key Laboratory of Bioreactor Engineering and Shanghai Key Laboratory of Chemical Biology, School of Pharmacy, East China University of Science and Technology, #268, 130 Meilong Road, 200237, Shanghai, People's Republic of China aut Xie Sheng State Key Laboratory of Bioreactor Engineering and Shanghai Key Laboratory of Chemical Biology, School of Pharmacy, East China University of Science and Technology, #268, 130 Meilong Road, 200237, Shanghai, People's Republic of China aut Li Hegeng Department of Oncology, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, National Clinical Research Centre for Traditional Chinese Medicine and Oncology, 725, South Wanping Road, 200032, Shanghai, People's Republic of China aut Tian Jianhui Department of Oncology, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, National Clinical Research Centre for Traditional Chinese Medicine and Oncology, 725, South Wanping Road, 200032, Shanghai, People's Republic of China aut Liu Jianwen State Key Laboratory of Bioreactor Engineering and Shanghai Key Laboratory of Chemical Biology, School of Pharmacy, East China University of Science and Technology, #268, 130 Meilong Road, 200237, Shanghai, People's Republic of China aut Cell Stress and Chaperones Springer Netherlands 18/2(2013-03-01), 203-213 1355-8145 18:2<203 2013 18 12192 https://doi.org/10.1007/s12192-012-0374-y text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s12192-012-0374-y text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Zheng Yuanhong State Key Laboratory of Bioreactor Engineering and Shanghai Key Laboratory of Chemical Biology, School of Pharmacy, East China University of Science and Technology, #268, 130 Meilong Road, 200237, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Le Vanminh State Key Laboratory of Bioreactor Engineering and Shanghai Key Laboratory of Chemical Biology, School of Pharmacy, East China University of Science and Technology, #268, 130 Meilong Road, 200237, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Cheng Zhuoan State Key Laboratory of Bioreactor Engineering and Shanghai Key Laboratory of Chemical Biology, School of Pharmacy, East China University of Science and Technology, #268, 130 Meilong Road, 200237, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Xie Sheng State Key Laboratory of Bioreactor Engineering and Shanghai Key Laboratory of Chemical Biology, School of Pharmacy, East China University of Science and Technology, #268, 130 Meilong Road, 200237, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Li Hegeng Department of Oncology, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, National Clinical Research Centre for Traditional Chinese Medicine and Oncology, 725, South Wanping Road, 200032, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Tian Jianhui Department of Oncology, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, National Clinical Research Centre for Traditional Chinese Medicine and Oncology, 725, South Wanping Road, 200032, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Liu Jianwen State Key Laboratory of Bioreactor Engineering and Shanghai Key Laboratory of Chemical Biology, School of Pharmacy, East China University of Science and Technology, #268, 130 Meilong Road, 200237, Shanghai, People's Republic of China aut NATIONALLICENCE 773 0- Cell Stress and Chaperones Springer Netherlands 18/2(2013-03-01), 203-213 1355-8145 18:2<203 2013 18 12192 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer