Open AccessApoptosis inducing activity of 4‐substituted coumarins from Calophyllum brasiliense in human leukaemia HL‐60 cells
Author(s) -
Ito Chihiro,
Murata Tomiyasu,
Itoigawa Masataka,
Nakao Keisuke,
Kaneda Norio,
Furukawa Hiroshi
Publication year - 2006
Publication title -
journal of pharmacy and pharmacology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 118
eISSN - 2042-7158
pISSN - 0022-3573
DOI - 10.1211/jpp.58.7.0013
Subject(s) - apoptosis , cytotoxicity , caspase , caspase 3 , cell culture , chemistry , chromatin , staining , caspase 9 , programmed cell death , biology , microbiology and biotechnology , biochemistry , in vitro , genetics , dna
With the objective of identifying anti‐tumour‐promoting agents, we carried out a primary screening of ten 4‐substituted coumarins isolated from Calophyllum brasiliense Camb. (Guttiferae), to determine the ability of these compounds to inhibit proliferation of the human leukaemia cell line HL‐60. Among the 4‐substituted coumarins isolated, calophyllolide (2) and mammea B/BB (3) showed significant cytotoxicity against HL‐60 cells. Fluorescence microscopy with Hoechst 33342 staining revealed that the percentage of apoptotic cells with fragmented nuclei and condensed chromatin increased in a time‐dependent manner after treatment with calophyllolide (2) or mammea B/BB (3). In addition, the activity of caspase‐9 and caspase‐3 was also enhanced in a time‐dependent manner upon treatment with the 4‐substituted coumarins 2 and 3. Caspase‐9 and caspase‐3 inhibitors suppressed apoptosis induced by 4‐substituted coumarins 2 and 3. These results suggest that calophyllolide (2) and mammea B/BB (3) induced apoptosis in HL‐60 cells through activation of the caspase‐9/caspase‐3 pathway, which is triggered by mitochondrial dysfunction.