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Published on: March 30, 2019
Identification of the p33(ING1)-regulated genes that include cyclin B1 and proto-oncogene DEK by using cDNA
Masato Takahashi1, Naohiko Seki, Toshinori Ozaki
1Division of Biochemistry, Chiba Cancer Center Research Institute, Chuoh-ku, Chiba 260-8717, Japan.
Abstract:
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.
Insights
The inhibitor of growth 1 (ING1) protein influences cell cycle arrest and apoptosis. This study reveals ING1 targets multiple genes, including DEK and cyclin B1, with some regulation dependent on p53.
Area of Science:
- Molecular Biology
- Cancer Research
- Gene Regulation
Background:
- The tumor suppressor p33(ING1) is crucial for cell cycle arrest and apoptosis.
- ING1 functions as a transcriptional cofactor, interacting with p53 and chromatin-modifying enzymes.
- The precise molecular mechanisms of ING1-mediated transcriptional regulation remain unclear.
Purpose of the Study:
- To investigate the molecular mechanisms of ING1-mediated transcriptional regulation.
- To identify genes regulated by ING1 in mammary epithelial cells.
- To determine the role of p53 in ING1-mediated gene expression changes.
Main Methods:
- cDNA microarray analysis of NMuMG cells with antisense ING1 overexpression.
- Semiquantitative reverse transcription-PCR for validating gene expression changes.
- Adenovirus-mediated ING1 overexpression and analysis in NMuMG and p53-deficient SAOS2 cells.
Main Results:
- Antisense ING1 overexpression upregulated 14 genes (e.g., cyclin B1, DEK) and downregulated 5 genes (e.g., TPT1).
- ING1 overexpression downregulated cyclin B1, DEK, and osteopontin, while increasing TPT1 expression.
- p33(ING1)-induced cyclin B1 downregulation was confirmed to be p53-dependent.
Conclusions:
- p33(ING1) targets multiple genes, including proto-oncogene DEK and cyclin B1, affecting cell growth and apoptosis.
- ING1-mediated transcriptional regulation involves both p53-dependent and independent pathways.
- This study elucidates key aspects of ING1's role in gene regulation and cellular processes.
