Cisplatin effects on F-actin and matrix proteins precede renal tubular cell detachment and apoptosis in vitro.
In primary cultures of porcine proximal tubular kidney cells and LLC-PK1 cells cisplatin (5 - 50 microM) caused apoptosis and cell detachment; in both systems cell detachment occurred, preceded by a loss of cytoskeletal F-actin stress fibers within 4 - 6 h, and a reduction of mRNA encoding for fibronectin, collagen a2 type (IV) and laminin B2 within 17 - 41 h. Prevention of F-actin damage by phalloidin prevented nuclear fragmentation, suggesting a relation between F-actin damage and apoptosis. Overexpression of Bcl-2 also prevented apoptosis, but did not prevent damage to the F-actin skeleton or the reduction of mRNA expression of the matrix proteins. These results suggest that Bcl-2 overexpression interferes with apoptotic signals downstream of F-actin. The relevance of these results for cell detachment in kidney toxicity is discussed.
Kruidering
M
M
Division of Toxicology, Leiden/Amsterdam Center for Drug Research, Leiden University, P.O. Box 9503, 2300 RA Leiden, The Netherlands.
van de Water
B
B
Zhan
Y
Y
Baelde
J J
JJ
Heer
E
E
Mulder
G J
GJ
Stevens
J L
JL
Nagelkerke
J F
JF
D000199:Actins / Q000187:drug effects* / Q000502:physiology; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002448:Cell Adhesion / Q000187:drug effects; D002467:Cell Nucleus; D002470:Cell Survival / Q000187:drug effects; D002945:Cisplatin / Q000494:pharmacology*; D003599:Cytoskeleton; D016326:Extracellular Matrix Proteins / Q000378:metabolism*; D015870:Gene Expression; D007687:Kidney Tubules, Proximal / Q000166:cytology*; D018374:LLC-PK1 Cells; D008870:Microtubules; D009336:Necrosis; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger; D013552:Swine
D000199:Actins; D016326:Extracellular Matrix Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D002945:Cisplatin