Daishiro Miura, Hiroshi Uno, Yoshiaki Azuma, Tomohiro Ohta*,#,
Mamoru Kiyoki and Yoshihiro Izawa
Teijin Institute for Bio-Medical Research, 4-3-2 Asahigaoka, Hino, Tokyo 191-8512, Japan
*╩To whom correspondence should be addressed.
#╩Present address for correspondence: Pharmaceuticals Scientific Sales Promotion Department, Teijin Ltd., Iino Bldg. 5F, 1-1, Uchisaiwaicho 2-chome, Chiyoda-ku, Tokyo 100-8585, Japan
Abstract: The effect of TEI-6363 (5-[E-4-N,╩N╩-dimethylaminophenylmethylene]-4-hydroxy-2-[1-methyl imidazole-2-ilthio]-4-[4-phenylbutyl]-2-cyclopentenone), a chemically synthesized prostaglandin╩A1 derivative, on cell proliferation and osteoblastic differentiation was investigated concurrently. ROS17/2.8 cells (a rat osteosarcoma-derived cell line) were treated with TEI-6363 at two concentrations, 10-7 and 10-6╩M, and viable cells were counted to assess cytotoxic effects and determine the growth curve. After 96╩h of treatment, there was no evidence of any effect of TEI-6363 on cell viability at either concentration. However, a clear inhibitory effect on cell proliferation was observed after treatment with 10-6╩M TEI-6363 for 24╩h or longer. A pulse-treatment experiment showed that TEI-6363 induced the inhibition of proliferating ROS17/2.8 cells 24╩h after addition. The inhibition of proliferation was associated with G1-arrest demonstrated by flow cytometric analysis, and incorporation of [3H]thymidine by ROS17/2.8 cells was decreased. Osteoblastic differentiation (assessed on the basis of increased alkaline phosphatase activity and collagen synthesis) was induced by TEI-6363 treatment at 10-6╩M following G1-arrest and inhibition of cell proliferation. These results suggest that TEI-6363 arrested the cell cycle of ROS17/2.8 cells at the G1╩phase and induced osteoblastic differentiation. These results did not appear to be dependent on a marked cytotoxic effect.
Keywords: Prostaglandin, Cyclopentenone, Osteoblast, G1 arrest, Differentiation