Alterations of the actin polymerization status as an apoptotic morphological effector in HL-60 cells.
The alterations of the cytoskeletal actin network have been implicated as a morphological effector in apoptosis. However, studies directly linking actin change to the morphological events in apoptosis are lacking. This study quantitatively examined the effect of actin alteration on the camptothecin (CPT)-induced apoptotic process in HL-60 cells. Actin alteration was induced by two distinctive types of agent: the polymerization-stimulating agent, Jasplakinolide (Jas), and the polymerization-blocking agent, cytochalasin B (CB). The actin polymerization status was measured by two complementary methods: the cell pellet-based DNase I inhibition method, and the individual cell-based quantitative fluorescence image analysis (QFIA) assay. Actin polymerization induced by Jas caused apoptosis directly. By contrast, CB, an actin polymerization-blocking agent, partially inhibited CPT-induced apoptosis. A similar inhibition of the CPT-induced apoptosis response was observed with a more specific actin depolymerization agent, cytochalasin E. The alterations of the actin polymerization status occurred in three sequential steps during the apoptotic process: first polymerization, followed by depolymerization, and finally degradation. However, compared with CPT-induced apoptosis, Jas-induced apoptosis was characterized by pronounced actin polymerization that corresponded morphologically with prominent membrane blebbing, but less apoptotic body formation. Furthermore, DNase I activity, which is normally inhibited by G-actin, was specifically detected in Jas-treated cells. These results show that the regulation of actin polymerization is an important apoptotic morphological effector, whereas the alterations of the actin polymerization status by chemicals have profound effects not only on altering the morphology of apoptotic cells, but on apoptosis induction in HL-60 cells as well.
Rao
J Y
JY
Department of Pathology and Laboratory Medicine, University of California at Los Angeles and Jonsson Comprehensive Cancer Center, Los Angeles, California 90024, USA.
Jin
Y S
YS
Zheng
Q
Q
Cheng
J
J
Tai
J
J
Hemstreet
G P
GP
D000199:Actins / Q000187:drug effects* / Q000378:metabolism*; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis; D001704:Biopolymers / Q000378:metabolism; D002166:Camptothecin / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects; D003571:Cytochalasin B / Q000494:pharmacology; D003572:Cytochalasins / Q000494:pharmacology; D003850:Deoxyribonuclease I / Q000378:metabolism; D047630:Depsipeptides; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D018922:HL-60 Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D020287:In Situ Nick-End Labeling; D008856:Microscopy, Fluorescence; D010456:Peptides, Cyclic / Q000494:pharmacology; D013997:Time Factors
D000199:Actins; D000970:Antineoplastic Agents; D001704:Biopolymers; D003572:Cytochalasins; D047630:Depsipeptides; D010456:Peptides, Cyclic; C057531:jasplakinolide; C007215:cytochalasin E; D003571:Cytochalasin B; D003850:Deoxyribonuclease I; D002166:Camptothecin