Fluorescence lifetime imaging of E-combretastatin uptake and distribution in live mammalian cells.
To investigate within live mammalian cells the uptake and disposition of combretastatins, fluorescence lifetime imaging was used with two-photon excitation (2PE). Combretastatin A4 (CA4) and analogues are potential anticancer drugs due to their ability to inhibit angiogenesis. E(trans)-combretastatins are considerably less active than the Z(cis)-combretastatins proposed for clinical use. However the E-combretastatins exhibit stronger intrinsic fluorescence with quantum yields and lifetimes that depend markedly on solvent polarity and viscosity. It is proposed that 2PE in the red and near-infrared tissue window may allow in situ isomerization of E-combretastatins to the more active Z-isomer, offering spatial and temporal control of drug activation and constitute a novel form of photodynamic therapy. In the present work we have characterised 2PE of E-CA4 and have used fluorescence lifetime imaging with 2PE to study uptake and intracellular disposition of E-CA4 and an analogue. The results show that these molecules accumulate rapidly in cells and are located mainly in lipidic environments such as lipid droplets. Within the droplets the local concentrations may be up to two orders of magnitude higher than that of the drug in the surrounding medium.
School of Environment and Life Sciences, University of Salford, Salford M5 4WT, UK. r.h.bisby@salford.ac.uk
Roger H
RH
Bisby
Stanley W
SW
Botchway
John A
JA
Hadfield
Alan T
AT
McGown
Anthony W
AW
Parker
Kathrin M
KM
Scherer
D000818:Animals; D001632:Bibenzyls / Q000737:chemistry / Q000493:pharmacokinetics*; D016466:CHO Cells; D006224:Cricetinae; D006367:HeLa Cells; D006801:Humans; D036641:Microscopy, Fluorescence, Multiphoton; D013267:Stilbenes / Q000737:chemistry / Q000493:pharmacokinetics*
D001632:Bibenzyls; D013267:Stilbenes; C040105:combretastatin; C058728:fosbretabulin