Post-translational modification of cyclin A1 is associated with staurosporine and TNFalpha induced apoptosis in leukemic cells.
Understanding of molecular mechanisms underlying the effects of cell cycle proteins in response to the chemotherapeutic agents is of great importance for improving the efficacy of targeted therapeutics and overcoming resistance to chemotherapeutic agents. Staurosporine and tumor necrosis factor alpha (TNFalpha) are the therapeutic agents that inhibit tumor cell growth by inducing cell death. Staurosporine induces apoptosis through the intrinsic pathway, while TNFalpha trigger the cell death via the extrinsic apoptotic pathway. We have previously demonstrated that the cell cycle regulatory protein, cyclin A1 played an important role in the development of acute myeloid leukemia (AML), and cyclin A1 expression correlated with disease characteristics and patient outcome in leukemia. However, it remains unknown how cyclin A1 expression is regulated in leukemic cells treated with the therapeutic agents. Here, we demonstrate that cyclin A1 protein is regulated by proteasome-mediated ubiquitination and degradation in untreated U-937 cells. Interestingly, ubiquitination- and proteasomal-mediated degradation of cyclin A1 is prevented in cells treated with staurosporine or TNFalpha. Induction of apoptosis in U-937 cells by staurosporine or TNFalpha resulted in an increase in cyclin A1 protein expression, which correlated well with cyclin A1 protein modification and the activation of caspase-3. Blocking caspases activity by Z-VAD-FMK had no effect on the increased cyclin A1 expression, suggesting that cyclin A1 might be regulated by caspase-3 independent pathways. We further propose that CDC25C may be associated with cyclin A1 protein modification in response to staurosporine or TNFalpha treatment. Our results suggest that cyclin A1 protein is stabilized via post-transcriptional modification in response to apoptosis induced by staurosporine or TNFalpha.
Ekberg
Jenny
J
Division of Pathology, Department of Laboratory Medicine, Lund University, Malmö University Hospital, 205 02 Malmo, Sweden.
Persson
Jenny Liao
JL
D000111:Acetylcysteine / Q000031:analogs & derivatives / Q000494:pharmacology; D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism; D061945:Caspase Inhibitors; D045744:Cell Line, Tumor / Q000187:drug effects / Q000378:metabolism; D056750:Cyclin A1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051357:Cyclin-Dependent Kinase 2 / Q000378:metabolism; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006801:Humans; D007938:Leukemia / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D061988:Proteasome Inhibitors; D011499:Protein Processing, Post-Translational / Q000187:drug effects*; D019311:Staurosporine / Q000494:pharmacology*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D054875:Ubiquitination; D020687:cdc25 Phosphatases / Q000378:metabolism
D000590:Amino Acid Chloromethyl Ketones; C503745:CCNA1 protein, human; D061945:Caspase Inhibitors; D056750:Cyclin A1; D015853:Cysteine Proteinase Inhibitors; D004791:Enzyme Inhibitors; D061988:Proteasome Inhibitors; D014409:Tumor Necrosis Factor-alpha; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; C067713:lactacystin; D051357:Cyclin-Dependent Kinase 2; D020687:cdc25 Phosphatases; D053148:Caspase 3; D046988:Proteasome Endopeptidase Complex; D019311:Staurosporine; D000111:Acetylcysteine
Int J Biochem Cell Biol. 2006;38(8):1330-9
16517207
J Biol Chem. 2001 Nov 30;276(48):45255-60
11572869
Mol Biol Cell. 1995 Feb;6(2):185-97
7787245
Nat Med. 1997 Feb;3(2):222-5
9018243
Nat Rev Cancer. 2007 Jul;7(7):495-507
17568790
Cell Growth Differ. 1996 Jun;7(6):699-710
8780883
Nat Genet. 1998 Nov;20(3):266-72
9806545
Genes Dev. 1999 Nov 1;13(21):2751-7
10557204
Methods Enzymol. 1991;201:340-7
1719345
Oncogene. 2004 Dec 2;23(56):9082-9
15489899
Nat Rev Mol Cell Biol. 2001 Jan;2(1):63-7
11413467
Int J Cancer. 1998 Mar 16;75(6):925-32
9506539
Oncogene. 2004 Mar 15;23(11):2009-15
15021888
Curr Opin Genet Dev. 2000 Feb;10(1):54-64
10679394
Proc Natl Acad Sci U S A. 2001 Jun 5;98(12):6853-8
11381140
Genes Dev. 1999 Feb 1;13(3):270-83
9990852
Eur J Haematol. 2005 Aug;75(2):106-15
16004607
Nature. 1991 Jan 10;349(6305):132-8
1846030
J Cell Physiol. 2001 Apr;187(1):1-10
11241344
FEBS Lett. 1997 Jun 23;410(1):96-106
9247131
Leuk Res. 2006 Mar;30(3):254-61
16182364
Blood. 1999 Mar 15;93(6):2067-74
10068680
EMBO J. 1996 Jan 2;15(1):46-54
8598205
Cancer Res. 1995 Jan 15;55(2):228-31
7812949
Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):3754-8
8170983
Oncogene. 2005 Apr 14;24(16):2739-44
15829981
Int J Oncol. 2003 Jan;22(1):15-20
12469180
J Biol Chem. 1999 Jan 8;274(2):1156-63
9873064
Cell Prolif. 2003 Jun;36(3):165-75
12814432
Mol Cell Biol. 1999 Mar;19(3):2400-7
10022926
Oncogene. 2000 Jun 22;19(27):3059-68
10871858
Cell. 1997 Oct 31;91(3):325-34
9363941
Neoplasia. 2000 Jul-Aug;2(4):291-9
11005563
FEBS Lett. 2001 Feb 16;490(3):117-22
11223026
Mol Cell Biol. 1998 May;18(5):2912-22
9566910
Nat Genet. 1998 Dec;20(4):377-80
9843212
Proc Natl Acad Sci U S A. 2003 Sep 30;100 Suppl 1:11842-9
14504387
J Biol Chem. 2001 Oct 26;276(43):39667-78
11485996
Oncogene. 2003 Dec 8;22(56):9048-57
14663483
Proc Natl Acad Sci U S A. 1992 Sep 15;89(18):8626-30
1528872
Genes Dev. 1999 Aug 1;13(15):1899-911
10444588