[An experimental study of bladder cancer cell apoptosis induced by cinobufacin].
OBJECTIVE
To study the mechanism of cinobufacin-induced apoptosis in BIU87 cell.


METHODS
Growth inhibition of bladder cancer cells in vitro were estimated using MTT assay. Apoptotic cells were detected by in-situ cell apoptosis detection kit, and confirmed by flow cytometry. The mRNA level affected by cinobufacin was determined with RT-PCR. Caspase-3 activity was estimated with chromometry.


RESULTS
Cinobufacin inhibited the proliferation of BIU87 cells and induced apoptosis. More than 50% cells were killed with 0.2 mg/L of cinobufacin for 72 h. S-phase and G2-phase arrest was induced. There were significant changes of Bcl-2, caspase-3 mRNA and caspase-3 activity; while no changes of bax was found.


CONCLUSIONS
Cinobufacin can inhibit the proliferation of BIU87 cells by inducting apoptosis, which may be related with S- and G2-phase arrest, down-regulation of Bcl-2, and increasing caspase-3 expression and its activity.
Dpartment of Urology, Affiliated Hospital of Guangdong Medical College, Zhanjiang 524000, China. sj366@21cn.com
Jin
J
Su
Zhi-jian
ZJ
Xu
Mu-shi
MS
Ye
D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry; D016195:G2 Phase / Q000187:drug effects; D020128:Inhibitory Concentration 50; D008856:Microscopy, Fluorescence; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016196:S Phase / Q000187:drug effects; D013997:Time Factors; D001749:Urinary Bladder Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D051028:bcl-2-Associated X Protein / Q000235:genetics / Q000378:metabolism
D000970:Antineoplastic Agents; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; D053148:Caspase 3