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KMID : 0380020070220030168
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Korean Journal of Biotechnology and Bioengineering
2007 Volume.22 No. 3 p.168 ~ p.173
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Fluorescence Detection for Protoporphyrin IX Induced from 5-ALA and ALA-methyl ester in Incubated Liver Cancer Cells
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Kim Myung-Hwa
Kim Jung-Mi
Kim Hyun-Jeong
Lee In-Seon
Kim Kyung-Chan
Lee Chang-Seop
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Abstract
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To clarify the usefulness of fluorescent diagnosis for cancer, we investigated the optimal method of administrating 5-aminolevulinic acid (5-ALA), 5-aminolevulinic acid methyl ester (ALA-methyl ester) by analyzing fluorescence signal of Protoporphyrin IX (PpIX) in the cultured normal and cancer cells. 5-ALA and ALA-methyl ester was injected as a photosensitizer to the cancer liver cells (HepG2) and normal liver cells (Chang). Chang and HepG2 cells were incubated with various concentrations of 5-ALA and ALA-methyl ester (0-800 μg/mL). The accumulation of PpIX induced by 5-ALA and ALA-methyl ester was in HepG2 and Chang. The cell viability was measured by MTT assay. Fluorescence of PpIX in HepG2 cell was excited at a wavelength (λ = 410 ㎚) and showed an emission spectrum at 603.2 nm, 660.8 nm and 603.2 nm, 661.4 nm which could be related to the PpIX generation induced by the applied 5-ALA and ALA-methyl ester, respectively. The experimental results showed that fluorescence signal of PpIX was proportional to the concentration of 5-ALA and ALA-methyl ester in tumor cells, but measured with low concentration in normal cells. Another results showed that the PpIX formation rate induced by ALA-methyl ester is higher than that of 5-ALA.
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KEYWORD
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5-aminolevulinic acid, 5-aminolevulinic acid methyl ester, protoporphyrin IX, photosensitizer
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