Lysosome imaging in cancer cells by pyrene-benzothiazolium dyes: An alternative imaging approach for LAMP-1 expression based visualization methods to avoid background interference |
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| Affiliation: | 1. Department of Chemistry, University of Akron, Akron, OH 44325, USA;2. Maurice Morton Institute of Polymer Science, University of Akron, Akron, OH 44325, USA;3. Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN 46556, USA;4. Department of Medicinal Chemistry and Molecular Pharmacology and the Purdue Center for Cancer Research, Purdue University, West Lafayette, IN 47907, USA;1. State Key Laboratory of Biotherapy and Cancer Center, Department of Orthopedics, West China Hospital, Collaborative Innovation Center for Biotherapy, Sichuan University, Chengdu, 610041, China;2. Key Laboratory of Birth Defects and Related Diseases of Women and Children, West China Second University Hospital, Sichuan University, Chengdu, 610041, China;3. Key Laboratory of Green Chemistry and Technology, Ministry of Education, College of Chemistry, Sichuan University, Chengdu, 610064, China;4. Chengdu Synguider Technology Co., Ltd., Chengdu, 610041, China;1. CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China;2. Fluorescence Research Group, Singapore University of Technology and Design, Singapore 487372, Singapore;3. University of Chinese Academy of Sciences, Beijing 100039, China;1. Key Laboratory of Green Chemistry and Technology (Ministry of Education), College of Chemistry, Sichuan University, Chengdu 610064, PR China;2. State Key Laboratory of Biotherapy/Collaborative Innovation Center for Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, PR China;3. State Key Laboratory of Oral Diseases, Sichuan University, Chengdu 610041, PR China;1. Department of Criminal Science and Technology, Sichuan Police College, Luzhou 646000, China;2. Liaoning Water Environment Monitoring Center, Shenyang Branch, Shenyang 110005, China;3. State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian 116024, China |
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| Abstract: | A series of pyrene-benzothiazolium dyes (1a–1d) were experimentally investigated to study their internalization mechanism into cellular lysosomes as well as their potential imaging applications for live cell imaging. The lysosome selectivity of the probes was further compared by using fluorescently tagged lysosome associated membrane protein-1 (LAMP-1) expression-dependent visualization in both normal (COS-7, HEK293) and cancer (A549, Huh 7.5) cell lines. These probes were successfully employed as reliable lysosome markers in tumor cell models, thus providing an attractive alternative to LAMP-1 expression-dependent visualization methods. One advantage of these probes is the elimination of significant background fluorescence arising from fluorescently tagged protein expression on the cell surface when cells were transfected with LAMP-1 expression plasmids. Probes exhibited remarkable ability to stain cellular lysosomes for long-term experiments (up to 24 h) and the highly lipophilic nature of the probe design allowed their accumulation in hydrophobic regions of the cellular lysosomes. Experimental evidences indicated that the probes are likely to be internalized into lysosomes via endocytosis and accumulated in the hydrophobic regions of the lysosomes rather than in the acidic lysosomal lumen. These probes also demonstrated significant stability and lysosome staining for fixed cell imaging applications as well. Lastly, the benzothiazolium moiety of the probes was identified as the key component for lysosome selectivity. |
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