Abstract
We have investigated the molecular mechanisms regulating the necrosis and apoptosis that occur on treatment of mouse mammary tumor FM3A cells with 5-fluoro-2′-deoxyuridine (FUdR), a potent anticancer agent, using the original clone F28-7 and its variant F28-7-A cells. Previously, we reported an interesting observation that FUdR induces a necrotic morphology in F28-7 but an apoptotic morphology in F28-7-A cells. We have now analyzed the protein expression profiles of these FUdR-induced necrosis and apoptosis. Thus, proteome analysis of these clones by two-dimensional gel electrophoresis and mass spectrometry showed that the cytoplasmic intermediate filament protein, cytokeratin-19, is expressed at a significantly higher level in F28-7 than in F28-7-A cells. This strong expression was detected both in untreated and FUdR-treated stages of F28-7 cells. We interpreted this phenomenon as suggesting that cytokeratin-19 possesses a function in leading the cell to apoptosis. We performed a knockdown of cytokeratin-19 expression in F28-7 cells by use of the small interfering RNA technique. Indeed, a lowering of the cytokeratin-19 expression down to the level in F28-7-A occurred, and the FUdR-induced death morphology of this knockdown F28-7 was apoptosis, instead of the necrosis usually observable in the FUdR-treated F28-7. It is known that the cytoskeletal protein cytokeratin-19 undergoes caspase-mediated degradation during apoptosis. Our present finding provides an interesting possibility that cytokeratin-19 may have a key role in regulating cell-death morphology.
| Original language | English |
|---|---|
| Pages (from-to) | 2329-2338 |
| Number of pages | 10 |
| Journal | Journal of Proteome Research |
| Volume | 9 |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - May 7 2010 |
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Keywords
- 5-fluoro-2′-deoxyuridine (FUdR)
- Apoptosis
- Cell death
- Cytokeratin-19
- Intermediate filament
- Lamin B1
- Necrosis
- Proteome analysis
- SiRNA
ASJC Scopus subject areas
- Biochemistry
- Chemistry(all)
Cite this
Protein expression profiles of necrosis and apoptosis induced by 5-fluoro-2′-deoxyuridine in mouse cancer cells. / Sato, Akira; Satake, Akito; Hiramoto, Akiko; Wataya, Yusuke; Kim, Hye-Sook.
In: Journal of Proteome Research, Vol. 9, No. 5, 07.05.2010, p. 2329-2338.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Protein expression profiles of necrosis and apoptosis induced by 5-fluoro-2′-deoxyuridine in mouse cancer cells
AU - Sato, Akira
AU - Satake, Akito
AU - Hiramoto, Akiko
AU - Wataya, Yusuke
AU - Kim, Hye-Sook
PY - 2010/5/7
Y1 - 2010/5/7
N2 - We have investigated the molecular mechanisms regulating the necrosis and apoptosis that occur on treatment of mouse mammary tumor FM3A cells with 5-fluoro-2′-deoxyuridine (FUdR), a potent anticancer agent, using the original clone F28-7 and its variant F28-7-A cells. Previously, we reported an interesting observation that FUdR induces a necrotic morphology in F28-7 but an apoptotic morphology in F28-7-A cells. We have now analyzed the protein expression profiles of these FUdR-induced necrosis and apoptosis. Thus, proteome analysis of these clones by two-dimensional gel electrophoresis and mass spectrometry showed that the cytoplasmic intermediate filament protein, cytokeratin-19, is expressed at a significantly higher level in F28-7 than in F28-7-A cells. This strong expression was detected both in untreated and FUdR-treated stages of F28-7 cells. We interpreted this phenomenon as suggesting that cytokeratin-19 possesses a function in leading the cell to apoptosis. We performed a knockdown of cytokeratin-19 expression in F28-7 cells by use of the small interfering RNA technique. Indeed, a lowering of the cytokeratin-19 expression down to the level in F28-7-A occurred, and the FUdR-induced death morphology of this knockdown F28-7 was apoptosis, instead of the necrosis usually observable in the FUdR-treated F28-7. It is known that the cytoskeletal protein cytokeratin-19 undergoes caspase-mediated degradation during apoptosis. Our present finding provides an interesting possibility that cytokeratin-19 may have a key role in regulating cell-death morphology.
AB - We have investigated the molecular mechanisms regulating the necrosis and apoptosis that occur on treatment of mouse mammary tumor FM3A cells with 5-fluoro-2′-deoxyuridine (FUdR), a potent anticancer agent, using the original clone F28-7 and its variant F28-7-A cells. Previously, we reported an interesting observation that FUdR induces a necrotic morphology in F28-7 but an apoptotic morphology in F28-7-A cells. We have now analyzed the protein expression profiles of these FUdR-induced necrosis and apoptosis. Thus, proteome analysis of these clones by two-dimensional gel electrophoresis and mass spectrometry showed that the cytoplasmic intermediate filament protein, cytokeratin-19, is expressed at a significantly higher level in F28-7 than in F28-7-A cells. This strong expression was detected both in untreated and FUdR-treated stages of F28-7 cells. We interpreted this phenomenon as suggesting that cytokeratin-19 possesses a function in leading the cell to apoptosis. We performed a knockdown of cytokeratin-19 expression in F28-7 cells by use of the small interfering RNA technique. Indeed, a lowering of the cytokeratin-19 expression down to the level in F28-7-A occurred, and the FUdR-induced death morphology of this knockdown F28-7 was apoptosis, instead of the necrosis usually observable in the FUdR-treated F28-7. It is known that the cytoskeletal protein cytokeratin-19 undergoes caspase-mediated degradation during apoptosis. Our present finding provides an interesting possibility that cytokeratin-19 may have a key role in regulating cell-death morphology.
KW - 5-fluoro-2′-deoxyuridine (FUdR)
KW - Apoptosis
KW - Cell death
KW - Cytokeratin-19
KW - Intermediate filament
KW - Lamin B1
KW - Necrosis
KW - Proteome analysis
KW - SiRNA
UR - http://www.scopus.com/inward/record.url?scp=77952055632&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=77952055632&partnerID=8YFLogxK
U2 - 10.1021/pr9010537
DO - 10.1021/pr9010537
M3 - Article
VL - 9
SP - 2329
EP - 2338
JO - Journal of Proteome Research
JF - Journal of Proteome Research
SN - 1535-3893
IS - 5
ER -