Open AccessColcemid‐induced apoptosis of cultured human glioma: Electron microscopic and confocal laser microscopic observation of cells sorted in different phases of cell cycle
Author(s) -
Tsuchida Takahiro,
Yoshimura Kunikazu,
Shirayama Yoshiaki,
Kawamoto Keiji
Publication year - 1998
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/(sici)1097-0320(19980401)31:4<295::aid-cyto9>3.0.co;2-i
Subject(s) - colcemid , interphase , mitosis , confocal microscopy , biology , microbiology and biotechnology , flow cytometry , apoptosis , confocal , cell cycle , biochemistry , optics , physics
The effect of the antitubulin agent colcemid on human glioma cells was investigated by sorting cells with different DNA content and subjecting them to confocal laser microscopy and transmission electron microscopy. The human glioma cell line U251MG was exposed to colcemid at a concentration of 0.05 μg for 16 h. Flow cytometric analysis revealed the accumulation of cells in S/G2M phase. Cells harvested from each of G0/G1 and S/G2M peaks were then analyzed by confocal laser microscopy and transmission electron microscopy. Confocal laser microscopy revealed that colcemid‐treated cells harvested from the G0/G1 peak contained mitotic and apoptotic cells in addition to interphase cells. Electron microscopy confirmed that colcemid‐treated cells in the G0/G1 peak had fragmented nuclei typical of apoptotic cells and mitotic cells with altered chromatin structure. Some mitotic cells obtained by mitotic shake‐off after treatment with colcemid showed DNA strand breaks defined by in situ nick end labeling. The present study indicates that mitotic as well as interphase apoptosis occurs in U251MG cells following colcemid treatment. Cytometry 31:295–299, 1998. © 1998 Wiley‐Liss, Inc.