Improving the use of 5-aminolevulinic acid (ALA)-induced protoporphyrin IX (PPIX) for the gastrointestinal tract by esterification--an in vitro study.
Possible approaches to improve the diagnostic and therapeutic effects of 5-aminolevulinic acid (ALA)-induced protoporphyrin IX (PPIX) are the esterification of ALA for enhanced uptake and the choice of wavelength for irradiation. The human colonic cell lines HT29 [G2] and CCD18 (fibroblasts) were incubated with 0.6 mM ALA, ALA-hexylester or -benzylester respectively, and for further assays with hypotaurine, in addition. PPIX-accumulation was analyzed by flow cytometry and fluorescence spectroscopy. PPIX formation kinetics were continuously recorded. Incubated cells were irradiated with an incoherent light source lambda = 400-700 nm or lambda = 590-700 nm, respectively. After PDT treatment, clonogenicity assays were performed to determine cell viability. Esterification leads to increased PPIX-accumulation, decreased time for production of detectable amounts of PPIX as well as increased response to PDT. Tumor specificity is always maintained or exceeds values for ALA alone. ALA enters the cells via beta transporter whereas esters by passive diffusion. Altering irradiation wavelengths showed the independence of wavelength rather than light dose. Results emphasize the role of heme metabolism for generating tumor specificity rather than the process of ALA-uptake, an important detail for future clinical application.
Krieg
R C
RC
National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. krieg@cber.fda.gov
Uihlein
D
D
Murthum
T
T
Endlicher
E
E
Hausmann
F
F
Messmann
H
H
Knuechel
R
R
D000622:Aminolevulinic Acid / Q000627:therapeutic use*; D002460:Cell Line; D003106:Colon / Q000187:drug effects; D004064:Digestive System / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D004307:Dose-Response Relationship, Radiation; D005434:Flow Cytometry; D006801:Humans; D007700:Kinetics; D008027:Light; D010778:Photochemotherapy / Q000379:methods; D011524:Protoporphyrins / Q000096:biosynthesis*; D013050:Spectrometry, Fluorescence; D013997:Time Factors; D014407:Tumor Cells, Cultured
D011524:Protoporphyrins; D000622:Aminolevulinic Acid; C028025:protoporphyrin IX