title: Intracellular localization of the radiation enhancer motexafin gadolinium using interferometric Fourier fluorescence microscopy.
abstract: Motexafin gadolinium (MGd) is a unique therapeutic agent that localizes in cancer cells and increases tumor response to ionizing radiation and certain chemotherapeutics. The in vitro intracellular localization, accumulation, and retention of MGd in murine EMT6 mammary sarcoma and Rif-1 fibrosarcoma cell lines were studied using interferometric Fourier fluorescence microscopy. MGd cellular uptake was semiquantified using its characteristic fluorescence emission band centered at 758 nm. Colocalization studies were performed using mitochondrial, endoplasmic reticulum, Golgi apparatus, nuclear, and lysosomal fluorescent organelle probes, and verified using interferometric Fourier spectroscopy. Cellular uptake was gradual and increased significantly with incubation time. MGd localized primarily within the lysosomes and endoplasmic reticulum, and to a lesser extent within the Golgi apparatus and mitochondria. Mitochondrial staining was increased in media without serum. No nuclear uptake was detected in the Rif-1 cells, but after 48 h nuclear uptake was observed in 15% of EMT6 cells. These results indicated that MGd accumulates within cytoplasmic compartments. The sustained intracellular localization of MGd may, in part, account for its unique radiation and chemotherapy enhancement properties. Interferometric Fourier fluorescence microscopy is a potentially powerful tool in delineating and verifying localization sites of therapeutic agents.
authors.affiliation: Pharmacyclics Inc., Sunnyvale, California, USA. KWoodburn@pcyc.com
authors.forename: K W
authors.initials: KW
authors.lastname: Woodburn
mesh_terms: D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002467:Cell Nucleus / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005456:Fluorescent Dyes; D006056:Golgi Apparatus / Q000378:metabolism; D008247:Lysosomes / Q000378:metabolism; D008665:Metalloporphyrins / Q000032:analysis* / Q000493:pharmacokinetics; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods; D008857:Microscopy, Interference; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D009368:Neoplasm Transplantation; D009374:Neoplasms, Experimental / Q000378:metabolism*; D010777:Photochemistry / Q000379:methods; D011838:Radiation-Sensitizing Agents / Q000032:analysis* / Q000493:pharmacokinetics; D012509:Sarcoma / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D014407:Tumor Cells, Cultured
chemical_list: D016895:Culture Media, Serum-Free; D005456:Fluorescent Dyes; D008665:Metalloporphyrins; D011838:Radiation-Sensitizing Agents; C102825:motexafin lutetium; C437683:motexafin gadolinium