Apoptosis in Human Oral Squamous Cell Carcinomas is Induced by 15-Deoxy-
12,14-Prostaglandin J2 but not by Troglitazone
K. Fukuchi1,
M. Date2,3,*,
Y. Azuma2,
M. Shinohara2,
H. Takahashi3, and
K. Ohura2
1 First Department of Oral and Maxillofacial Surgery, Osaka Dental University, Hirakata, Japan;
2 Department of Pharmacology, Osaka Dental University, 8-1, Kuzuhahanazono-cho, Hirakata, Osaka 573-1121, Japan; and
3 Institute of Clinical Medicine and Research, Jikei University School of Medicine, Kashiwa, Japan;
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Figure 1. PPAR- protein expression in SAS and HSC-4 cells. (A) Western blot analysis. (B) Immunocytochemistry. Results shown are from one representative experiment from a total of 3 performed. Scale bar = 10 µm.
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Figure 2. Effects of 15-d-PGJ2 and troglitazone on cell viability. Cells were incubated with various concentrations of 15-d-PGJ2 and trogiltazone for 36 hrs (A) and with 20 µM 15-d-PGJ2 and troglitazone for the indicated periods of time (B). Cell viability was determined by a modified MTT assay and expressed as a percentage of viability under control culture conditions. Data are expressed as mean ± SD (n = 3). *p < 0.05, **p < 0.01.
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Figure 3. Detection of apoptosis in human oral SCC cell lines. (A) TUNEL staining in human oral SCC cell lines. A TUNEL assay was performed 24 hrs after 20 µM of 15-d-PGJ2 or troglitazone treatment. The results shown are from a representative experiment from a total of 3 that were performed. Scale bar = 10 µm. (B) Effect of 15-d-PGJ2 or troglitazone on intracellular nucleosome enrichment. The nucleosome concentration after 20 µM of 15-d-PGJ2 or troglitazone treatment was examined with a cell-death detection kit for the indicated periods of time. Control cells were assigned a value of 1, and other values were expressed relative to these and were plotted against the time after treatment. Data are expressed as mean ± SD (n = 3). *p < 0.05, **p < 0.01.
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Figure 4. Effects on apoptotic signaling pathways by 15-d-PGJ2. Control cells were assigned a value of 1, and other values were expressed relative to these and were plotted against the time after 15-d-PGJ2 treatment. Data are expressed as mean ± SD (n = 3). *p < 0.05, **p < 0.01. (A) Caspase activation induced by 15-d-PGJ2. Cells were incubated with 20 µM 15-d-PGJ2 and troglitazone for the indicated periods of time. (B) Effects of caspase inhibitors on 15-d-PGJ2-induced apoptosis. Cells were treated with 20 µM 15-d-PGJ2 in the presence or absence of Z-VAD-FMK (2.5-20 µM), or Z-DEVD-FMK (2.5-20 µM) for 24 hrs. Then cells were processed for detection of fragmented mono- and oligonucleosomes by ELISA. (C) Cytochrome c release from mitochondria. Cells were incubated with 20 µM 15-d-PGJ2 for the indicated periods of time.
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