Identification of the p33(ING1)-regulated genes that include cyclin B1 and proto-oncogene DEK by using cDNA microarray in a mouse mammary epithelial cell line NMuMG.
The candidate tumor suppressor p33(ING1) plays an important role in inducinggrowth arrest at G(0)-G(1) phase of the cell cycle and/or promoting apoptosis in cancerous cells. p33(ING1) is reported to act as a transcriptional cofactor by associating with tumor suppressor p53, HAT, or histone deacetyltransferase, suggesting that p33(ING1) is involved in chromatin-mediated transcriptional regulation. However, the molecular mechanism of p33(ING1)-mediated transcriptional regulation is poorly understood. Here we analyzed expression profiles in mouse mammary epithelial cells (NMuMG) by using a cDNA microarray consisting of 2304 mouse cDNAs after inducing transformation with antisense inhibitor of growth 1 (ING1) in retrovirus vector. The subsequent confirmation of the altered expression levels of the selected genes by semiquantitative reverse transcription-PCR demonstrated that overexpression of the antisense ING1 stimulated expression of 14 genes, which included cyclin B1, 12-O-tetradecanoylphorbol-13-acetate-inducible sequence 11, proto-oncogene DEK, and osteopontin, whereas we have detected transcriptional repression of 5 genes, including TPT1. In addition, adenovirus-mediated overexpression of ING1 in NMuMG cells resulted in down-regulation of cyclin B1, 12-O-tetradecanoylphorbol-13-acetate-inducible sequence 11, DEK, and osteopontin, whereas the levels of TPT1 expression were increased. The further analysis using p53(-/-) SAOS2 cells showed that the p33(ING1)-induced cyclin B1 down-regulation was p53 dependent. Thus, our cDNA microarray analysis suggested that p33(ING1) targets the multiple genes, including proto-oncogene DEK and cyclin B1, at least some of which are regulated in a p53-dependent manner, in the cells undergoing cell growth or apoptosis.
Takahashi
Masato
M
Division of Biochemistry, Chiba Cancer Center Research Institute, Chuoh-ku, Chiba 260-8717, Japan.
Seki
Naohiko
N
Ozaki
Toshinori
T
Kato
Masaki
M
Kuno
Tomoko
T
Nakagawa
Takahito
T
Watanabe
Ken-ichi
K
Miyazaki
Koh
K
Ohira
Miki
M
Hayashi
Shunji
S
Hosoda
Mitsuchika
M
Tokita
Hisashi
H
Mizuguchi
Hiroyuki
H
Hayakawa
Takao
T
Todo
Satoru
S
Nakagawara
Akira
A
D000818:Animals; D018797:Cell Cycle Proteins; D002460:Cell Line; D019926:Cyclin B / Q000096:biosynthesis / Q000235:genetics*; D056744:Cyclin B1; D016373:DNA, Antisense / Q000235:genetics / Q000494:pharmacology; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D004847:Epithelial Cells / Q000378:metabolism / Q000502:physiology; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D016147:Genes, Tumor Suppressor; D006131:Growth Inhibitors / Q000235:genetics / Q000502:physiology; D006801:Humans; D000076023:Inhibitor of Growth Protein 1; D047908:Intracellular Signaling Peptides and Proteins; D008321:Mammary Glands, Animal / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D051379:Mice; D009687:Nuclear Proteins; D020411:Oligonucleotide Array Sequence Analysis; D011506:Proteins / Q000235:genetics / Q000502:physiology*; D000090063:Proto-Oncogene Mas; D020794:Receptor Protein-Tyrosine Kinases / Q000096:biosynthesis / Q000235:genetics*; D036081:Receptors, Eph Family; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured; D000090324:Tumor Protein, Translationally-Controlled 1; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology; D025521:Tumor Suppressor Proteins
C534699:CCNB1 protein, human; C534700:Ccnb1 protein, mouse; D018797:Cell Cycle Proteins; C503698:CycB protein, Drosophila; D019926:Cyclin B; D056744:Cyclin B1; D016373:DNA, Antisense; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D006131:Growth Inhibitors; C501503:ING1 protein, human; C501505:Ing1 protein, mouse; D000076023:Inhibitor of Growth Protein 1; D047908:Intracellular Signaling Peptides and Proteins; C000715470:MAS1 protein, human; D009687:Nuclear Proteins; D011506:Proteins; D000090063:Proto-Oncogene Mas; C000715647:TPT1 protein, human; C000715648:Tpt1 protein, mouse; D000090324:Tumor Protein, Translationally-Controlled 1; D016159:Tumor Suppressor Protein p53; D025521:Tumor Suppressor Proteins; C119557:Dek protein, Drosophila; D020794:Receptor Protein-Tyrosine Kinases; D036081:Receptors, Eph Family