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Therapeutic effect of photodynamic therapy using PAD‐S31 and diode laser on human liver cancer cells
Author(s) -
Date Masataka,
Fukuchi Kazuhide,
Namiki Yoshihisa,
Okumura Akihiko,
Morita Shosuke,
Takahashi Hiroshi,
Ohura Kiyoshi
Publication year - 2004
Publication title -
liver international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.873
H-Index - 110
eISSN - 1478-3231
pISSN - 1478-3223
DOI - 10.1111/j.1478-3231.2004.00902.x
Subject(s) - photodynamic therapy , apoptosis , tunel assay , mtt assay , cancer research , cytochrome c , viability assay , microbiology and biotechnology , in vivo , photosensitizer , cancer cell , terminal deoxynucleotidyl transferase , programmed cell death , cell culture , in vitro , liver cancer , chemistry , cancer , medicine , biology , hepatocellular carcinoma , biochemistry , genetics , organic chemistry
Background/aims: Photodynamic therapy (PDT) is an effective local cancer treatment which a photosensitizer is administered and the tumor is irradiated with light. We examined the effect of PDT using PAD‐S31 as the photosensitizer, and the 670 nm diode laser on human hepatocellular carcinomas (HCCs). Methods: Huh‐7, HepG2 and Hep3B cell lines were used in the all experiments. Cell viability was determined by a modified MTT assay. Two methods were used for the determination of apoptosis: terminal deoxynucleotidyl transferase (TdT)‐mediated dUTP nick‐end labeling assay and detection of fragmented mono‐ and oligo‐nucleosomes by enzyme‐linked immunosorbent assay. The caspase activity was measured by fluorometric assay. Cytochrome c in cytosolic fraction was determined using a human cytochrome c immunoassay. Xenografts of human oral HCC cells were generated in KSN S1c nude mice. Results:In vitro PDT showed excellent cytotoxicity that was a function of laser energy, drug concentration and time to the hepatoma cell lines. The combined use of PAD‐S31 and laser irradiation showed excellent anti‐tumor activity without severe side‐effect against human hepatoma xenografts in nude mice. PDT‐mediated cell death occurred predominantly by apoptosis in vitro and in vivo . Furthermore, this treatment initiates early cytochrome c release, followed by late caspase‐3 and ‐9 activation. Conclusion: Our study demonstrates that PDT using PAD‐S31 and the diode laser induces apoptosis that is mediated by cytochrome c release and caspase activation in human liver cancer cell lines. It is expected that this therapy will be clinically useful for the treatment of patients with HCC.