Specific cytoskeleton changes during apoptosis accompanying induced differentiation of HL-60 myeloid leukemia cells.
Changes in actin filaments and microtubules were studied in the human myeloid leukemia HL-60 cell line during the process of apoptotic cell death accompanying induced differentiation. These cytoskeleton changes were assessed during a 6-day cultivation in the presence of 10(-6) M all-trans retinoic acid (ATRA), a specific inductor of both differentiation into granulocytes and apoptosis, or during a 18-day cultivation in the presence of 1.6 nM phorbol myristylacetate (PMA), which induces differentiation into macrophages. The processes were studied at the morphological level by fluorescence microscopy and, quantitatively, by flow cytometry. The results showed that the actin cytoskeleton underwent specific structural changes during the apoptotic process, but microtubules were not actively involved. In the initial stages of apoptosis, a fine meshwork of actin filaments turned into actin granules that, in the final stages, were transformed into a network of long actin fibres distributed throughout the cytoplasm. These actin structures were considered to play an active role in two main morphological events of apoptosis - formation of blebs and final cell disintegration into apoptotic bodies. In addition, high proportion of cells with apoptotic nuclei and completely destroyed actin structures were found in the differentiating ATRA-treated cell population. Flow cytometric measurement of cytoskeletal proteins content confirmed all these observed changes. Alterations and rearrangements of both cytoskeletal structures are common for the apoptotic cell death of HL-60 cells and they are independent on the course of differentiation.
Veselská
Renata
R
Cell Culture Laboratory, Department of Biology, Medical Faculty, Masaryk University, CZ-66243 Brno, Czech Republic. rvesel@med.muni.cz
Zitterbart
Karel
K
Jelínková
Sarka
S
Neradil
Jakub
J
Svoboda
Augustin
A
D000199:Actins / Q000378:metabolism*; D017209:Apoptosis / Q000187:drug effects*; D002454:Cell Differentiation / Q000187:drug effects*; D003599:Cytoskeleton / Q000187:drug effects*; D005434:Flow Cytometry; D006098:Granulocytes / Q000187:drug effects; D018922:HL-60 Cells; D006801:Humans; D007962:Leukocytes / Q000166:cytology / Q000187:drug effects; D008264:Macrophages / Q000187:drug effects; D008856:Microscopy, Fluorescence; D022423:Myeloid Cells / Q000166:cytology / Q000187:drug effects; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014212:Tretinoin / Q000494:pharmacology*
D000199:Actins; D014212:Tretinoin; D013755:Tetradecanoylphorbol Acetate