Apoptotic induction by BE16627B on human malignant glioma cell lines by an anti-matrix metalloproteinase agent.
We have elucidated the pharmacological action of the anti-matrix metalloproteinase inhibitor BE16627B on glioma cells. The study was limited to the noncytotoxic dose range. The aim of the study was to investigate whether the cytotoxicity of BE16627B, an anti-MMP agent, is related to apoptosis in the human malignant glioma cell lines U87MG, U251MG, and U373MG. MTT assay was performed to detect the cytotoxic dose range. Agarose gel electrophoresis was performed with purified genomic DNA following exposure to 20 to 500 microM BE16627B for 24 h, compared with 0 microM for the control group. Transmission electron microscopy (TEM) was employed to study nuclear fragmentation following exposure to 0, 20, and 500 microM of the agent for 24 h. An in situ endolabeling assay was performed to determine the index of apoptotic induction. MTT assay revealed that concentrations of 100 microM and above were cytotoxic. DNA laddering was demonstrated in agarose gel electrophoresis. TEM disclosed condensing and fragmentation of the chromatin. None of these changes were observed in the control group and the noncytotoxic dose group. The in situ endolabeling study disclosed that the apoptotic index was significantly elevated by cytotoxic doses of this agent (U373MG; control, 4.0%; 500 microM, 68.5%). These results indicated that cytotoxic concentrations of BE16627B induced apoptosis in human malignant glioma cell lines. In our previous report, this agent inhibited activity of MMP in noncytotoxic concentrations. Further study should be done to determine the pharmacological action of toxic BE16627B.
Yoshida
Daizo
D
Department of Neurosurgery, Nippon Medical School, 1-1-5 Sendagi. Bunkyo-ku, Tokyo 113-8603, Japan.
Watanabe
Kunihiro
K
Takahashi
Hiroshi
H
Sugisaki
Yuichi
Y
Teramoto
Akira
A
D017209:Apoptosis / Q000187:drug effects*; D045744:Cell Line, Tumor; D053938:DNA Fragmentation / Q000187:drug effects; D004151:Dipeptides / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004587:Electrophoresis, Agar Gel; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005910:Glioma / Q000378:metabolism / Q000473:pathology* / Q000648:ultrastructure; D006801:Humans; D061965:Matrix Metalloproteinase Inhibitors; D020782:Matrix Metalloproteinases / Q000187:drug effects*; D008854:Microscopy, Electron; D013386:Succinates / Q000494:pharmacology*
D004151:Dipeptides; D004791:Enzyme Inhibitors; D061965:Matrix Metalloproteinase Inhibitors; D013386:Succinates; C085667:L-N-(N-hydroxy-2-isobutylsuccinamoyl)seryl-L-valine; D020782:Matrix Metalloproteinases