Novel regulation of nuclear factor-YB by miR-485-3p affects the expression of DNA topoisomerase IIα and drug responsiveness.
Nuclear factor (NF)-YB, a subunit of the transcription factor nuclear factor Y (NF-Y) complex, binds and activates CCAAT-containing promoters. Our previous work suggested that NF-YB may be a mediator of topoisomerase IIα (Top2α), working through the Top2α promoter. DNA topoisomerase II (Top2) is an essential nuclear enzyme and the primary target for several clinically important anticancer drugs. Our teniposide-resistant human lymphoblastic leukemia CEM cells (CEM/VM-1-5) express reduced Top2α protein compared with parental CEM cells. To study the regulation of Top2α during the development of drug resistance, we found that NF-YB protein expression is increased in CEM/VM-1-5 cells compared with parental CEM cells. This further suggests that increased NF-YB may be a negative regulator of Top2α in CEM/VM-1-5 cells. We asked what causes the up-regulation of NF-YB in CEM/VM-1-5 cells. We found by microRNA profiling that hsa-miR-485-3p is lower in CEM/VM-1-5 cells compared with CEM cells. MicroRNA target prediction programs revealed that the 3'-untranslated region (3'-UTR) of NF-YB harbors a putative hsa-miR-485-3p binding site. We thus hypothesized that hsa-miR-485-3p mediates drug responsiveness by decreasing NF-YB expression, which in turn negatively regulates Top2α expression. To test this, we overexpressed miR-485-3p in CEM/VM-1-5 cells and found that this led to reduced expression of NF-YB, a corresponding up-regulation of Top2α, and increased sensitivity to the Top2 inhibitors. Results in CEM cells were replicated in drug-sensitive and -resistant human rhabdomyosarcoma Rh30 cells, suggesting that our findings represent a general phenomenon. Ours is the first study to show that miR-485-3p mediates Top2α down-regulation in part by altered regulation of NF-YB.
Department of Biopharmaceutical Sciences, University of Illinois at Chicago, Chicago, IL 60612, USA.
Cheng-Fen
CF
Chen
Xiaolong
X
He
Ahmet Dirim
AD
Arslan
Yin-Yuan
YY
Mo
William C
WC
Reinhold
Yves
Y
Pommier
William T
WT
Beck
D000951:Antigens, Neoplasm / Q000096:biosynthesis* / Q000235:genetics; D000970:Antineoplastic Agents / Q000633:toxicity; D023081:CCAAT-Binding Factor / Q000378:metabolism*; D045744:Cell Line, Tumor; D004250:DNA Topoisomerases, Type II / Q000096:biosynthesis* / Q000235:genetics; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics; D015536:Down-Regulation / Q000187:drug effects; D019008:Drug Resistance, Neoplasm; D005047:Etoposide / Q000633:toxicity*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D035683:MicroRNAs / Q000502:physiology*; D000075223:Poly-ADP-Ribose Binding Proteins; D013713:Teniposide / Q000633:toxicity; D015854:Up-Regulation / Q000187:drug effects
D000951:Antigens, Neoplasm; D000970:Antineoplastic Agents; D023081:CCAAT-Binding Factor; D004268:DNA-Binding Proteins; C548912:MIRN483 microRNA, human; D035683:MicroRNAs; D000075223:Poly-ADP-Ribose Binding Proteins; C402244:nuclear factor Y; D005047:Etoposide; D013713:Teniposide; D004250:DNA Topoisomerases, Type II; C000618811:TOP2A protein, human
Chem Biol. 2010 May 28;17(5):421-33
20534341
Proc Natl Acad Sci U S A. 2006 Jun 13;103(24):9136-41
16754881
J Immunol Methods. 1983 Dec 16;65(1-2):55-63
6606682
Nat Rev Cancer. 2009 May;9(5):338-50
19377506
Cancer Res. 2003 Dec 15;63(24):8634-47
14695175
J Biol Chem. 2000 Feb 11;275(6):4435-44
10660616
Cancer Res. 1989 Mar 1;49(5):1095-8
2563671
BMC Genomics. 2009 Jun 23;10:277
19549304
Cancer Cell. 2006 Jun;9(6):435-43
16766263
Cell. 2004 Jan 23;116(2):281-97
14744438
Oncogene. 2008 Jun 19;27(27):3880-8
18264139
Nucleic Acids Res. 1995 Nov 25;23(22):4565-72
8524643
Biochemistry. 1988 Nov 29;27(24):8861-9
2853972
Cancer Res. 2003 Dec 1;63(23):8167-72
14678971
Proc Natl Acad Sci U S A. 1989 Dec;86(23):9431-5
2556712
Oncogene. 2008 Jan 10;27(3):378-86
17621267
Blood. 2002 Mar 15;99(6):2185-91
11877296
Biochem J. 2003 Sep 15;374(Pt 3):723-9
12769819
Mol Pharmacol. 2000 Feb;57(2):296-307
10648639
Nat Rev Mol Cell Biol. 2002 Jun;3(6):430-40
12042765
Mol Cell Biol. 2008 Sep;28(17):5147-61
18573883
Eur J Cancer. 1996 Jun;32A(6):958-66
8763336
Nat Rev Cancer. 2009 May;9(5):327-37
19377505
Cell Res. 2009 Apr;19(4):439-48
19238171
Nat Rev Cancer. 2006 Apr;6(4):259-69
16557279
Proc Natl Acad Sci U S A. 1999 Jul 6;96(14):7859-64
10393912
J Biol Chem. 2001 Feb 23;276(8):5570-6
11096075
Mol Pharmacol. 2001 Feb;59(2):203-11
11160854
Proc Natl Acad Sci U S A. 1988 Oct;85(19):7177-81
2845399
Nat Genet. 2009 Mar;41(3):365-70
19219043
Science. 2003 Jul 18;301(5631):336-8
12869753
Annu Rev Biochem. 1989;58:351-75
2549853
J Biol Chem. 2009 Sep 25;284(39):26533-46
19633292
Proc Natl Acad Sci U S A. 2007 Aug 14;104(33):13513-8
17686970
Nat Rev Cancer. 2006 Nov;6(11):857-66
17060945
Cancer Res. 1996 Jun 1;56(11):2573-83
8653700