Uropathogenic Escherichia coli toxins induce caspase-independent apoptosis in renal proximal tubular cells via ERK signaling.
BACKGROUND
Pyelonephritis is a risk factor for renal tubular epithelial cell damage. Recent studies have shown that Escherichia coli and/or its toxins may stimulate apoptotic cell death in renal tubular cells, but the underlying molecular mechanisms remain to be elucidated.


METHODS
Confluent LLC-PK(1) cells were exposed to E. coli toxins from overnight cultures of the uropathogenic O6K13H1 (O6) and the nonpathogenic W3110. The cell death was studied with morphological and biological assay.


RESULTS
E. coli soluble toxins from uropathogenic O6:K13:H1(O6) strain were found to induce apoptosis in a dose- and time-dependent manner in LLC-PK1 cells. The expression of FasR and the phosphorylation of ERK1/2 were significantly upregulated by O6 soluble toxins in a time-dependent manner. Cell death was completely inhibited by two specific ERK1/2 inhibitors, but not by a broad caspase inhibitor, zVAD-fmk, implicating a caspase-independent pathway via ERK. Moreover, we found that lysophosphatidic acid could trigger a survival signal through G-proteins and PI3K.


CONCLUSIONS
We demonstrate that apoptosis induced by uropathogenic E. coli toxins is dependent on ERK1/2. Caspases, although being activated, are not necessary for cell death, and they act after the ERK signaling at which point cells become committed to cell death or can be rescued.
Chen
Ming
M
Pediatric Nephrology Unit, Pathology and Immunology, Division of Pathology, Karolinska Institutet, Stockholm, Sweden. Ming.Chen@klinvet.ki.se
Jahnukainen
Timo
T
Bao
Wenjie
W
Daré
Elisabetta
E
Ceccatelli
Sandra
S
Celsi
Gianni
G
D000704:Analysis of Variance; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001427:Bacterial Toxins / Q000494:pharmacology*; D015153:Blotting, Western; D020169:Caspases / Q000502:physiology*; D002478:Cells, Cultured; D004926:Escherichia coli; D007687:Kidney Tubules, Proximal / Q000187:drug effects* / Q000201:enzymology; D020928:Mitogen-Activated Protein Kinases / Q000502:physiology*; D011704:Pyelonephritis / Q000201:enzymology / Q000473:pathology*; D015398:Signal Transduction; D013552:Swine; D015854:Up-Regulation
D001427:Bacterial Toxins; D020928:Mitogen-Activated Protein Kinases; D020169:Caspases