Susceptibility of hepatoma-derived cells to histone deacetylase inhibitors is associated with ID2 expression

Tsunedomi,Ryouichi; Iizuka,Norio; Harada,Sawako; Oka,Masaaki

doi:10.3892/ijo.2013.1811

Susceptibility of hepatoma-derived cells to histone deacetylase inhibitors is associated with ID2 expression

Authors:
- Ryouichi Tsunedomi
- Norio Iizuka
- Sawako Harada
- Masaaki Oka
View Affiliations

Affiliations: Department of Digestive Surgery and Surgical Oncology, Yamaguchi University Graduate School of Medicine, Ube, Yamaguchi 755-8505, Japan, Complementary Medicine, Yamaguchi University Graduate School of Medicine, Ube, Yamaguchi 755-8505, Japan
Published online on: February 6, 2013 https://doi.org/10.3892/ijo.2013.1811
Pages: 1159-1166
Copyright: © Tsunedomi et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Downregulation of inhibitor of DNA binding 2 (ID2) is associated with poor prognosis in cases of hepatocellular carcinoma (HCC). Therefore, to search for effective antitumor drugs for the treatment of HCC exhibiting poor prognostic indicators, we used two HCC-derived cell lines (HuH-7 and HLE) to alter ID2 levels. Specifically, ID2 expression was knocked down in HuH-7 cells via transfection with ID2-specific small interfering RNAs and separately ID2 was overexpressed in HLE cells via an ID2 expression plasmid vector. To assess the effect of antitumor drugs, MTS assay was performed. Annexin V staining was used to evaluate apoptosis and real-time RT-PCR was used to measure mRNA levels. ID2 knockdown cells were more susceptible to histone deacethylase (HDAC) inhibitors including sodium butyrate (NaB), sodium 4-phenyl-butyrate, tricostatin A, suberoylanilide hydroxamic acid, MS-275, apicidin and HC-toxin. Conversely, cells that overexpressed ID2 were less susceptible than control cells to HDAC inhibitors. NaB-induced apoptosis was inversely correlated with ID2 expression. Expression of the anti-apoptotic mRNA BCL2 was induced by NaB in control cells, but this induction of BCL2 was inhibited by ID2 knockdown and strengthened by ID2 overexpression. Expression of another anti-apoptotic mRNA, BCL2L1, was decreased by NaB administration and then partially recovered. However, in ID2 knockdown cells, BCL2L1 levels did not recover from NaB-induced suppression. ID2 affected the susceptibility of two HCC-derived cell lines to an HDAC inhibitor by regulating the expression of anti-apoptotic genes. Therefore, HDAC inhibitors may be effective for the treatment of HCC for which the prognosis is poor based on ID2 downregulation and ID2 could serve as a marker that is predictive of the clinical response to HDAC inhibitors.

View Figures

View References

1	Thorgeirsson SS and Grisham JW: Molecular pathogenesis of human hepatocellular carcinoma. Nat Genet. 31:339–346. 2002. View Article : Google Scholar : PubMed/NCBI
2	Parkin DM, Bray F, Ferlay J and Pisani P: Global cancer statistics. CA Cancer J Clin. 2005.55:74–108. 2002. View Article : Google Scholar : PubMed/NCBI
3	Bruix J, Boix L, Sala M and Llovet JM: Focus on hepatocellular carcinoma. Cancer Cell. 5:215–219. 2004. View Article : Google Scholar
4	Iizuka N, Oka M, Yamada-Okabe H, Mori N, Tamesa T, Okada T, Takemoto N, Sakamoto K, Hamada K, Ishitsuka H, Miyamoto T, Uchimura S and Hamamoto Y: Self-organizing-map-based molecular signature representing the development of hepatocellular carcinoma. FEBS Lett. 579:1089–1100. 2005. View Article : Google Scholar : PubMed/NCBI
5	Damdinsuren B, Nagano H, Kondo M, Yamamoto H, Hiraoka N, Yamamoto T, Marubashi S, Miyamoto A, Umeshita K, Dono K, Nakamori S, Wakasa K, Sakon M and Monden M: Expression of Id proteins in human hepatocellular carcinoma: relevance to tumor dedifferentiation. Int J Oncol. 26:319–327. 2005.PubMed/NCBI
6	Tsunedomi R, Iizuka N, Yamada-Okabe H, Tamesa T, Okada T, Sakamoto K, Takashima M, Hamaguchi T, Miyamoto T, Uchimura S, Hamamoto Y, Yamada M and Oka M: Identification of ID2 associated with invasion of hepatitis C virus-related hepatocellular carcinoma by gene expression profile. Int J Oncol. 29:1445–1451. 2006.PubMed/NCBI
7	Tsunedomi R, Iizuka N, Tamesa T, Sakamoto K, Hamaguchi T, Somura H, Yamada M and Oka M: Decreased ID2 promotes metastatic potentials of hepatocellular carcinoma by altering secretion of vascular endothelial growth factor. Clin Cancer Res. 14:1025–1031. 2008. View Article : Google Scholar : PubMed/NCBI
8	Benezra R, Davis R, Lockshon D, Turner D and Weintraub H: The protein ID: a negative regulator of helix-loop-helix DNA binding proteins. Cell. 61:49–59. 1990. View Article : Google Scholar : PubMed/NCBI
9	Kadesch T: Consequences of heteromeric interactions among helix-loop-helix proteins. Cell Growth Differ. 4:49–55. 1993.PubMed/NCBI
10	Norton JD: ID helix-loop-helix proteins in cell growth, differentiation and tumorigenesis. J Cell Sci. 113:3897–3905. 2000.PubMed/NCBI
11	Biggs J, Murphy EV and Israel MA: Id-like helix-loop-helix protein expressed during early development. Proc Natl Acad Sci USA. 89:1512–1516. 1992. View Article : Google Scholar : PubMed/NCBI
12	Hara E, Yamaguchi T, Nojima H, Ide T, Campisi J, Okayama H and Oda K: Id-related genes encoding helix-loop-helix proteins are required for G1 progression and are repressed in senescent human fibroblasts. J Biol Chem. 269:2139–2145. 1994.PubMed/NCBI
13	Rivera R and Murre C: The regulation and function of the Id proteins inlymphocyte development. Oncogene. 20:8308–8316. 2001. View Article : Google Scholar : PubMed/NCBI
14	Zebedee Z and Hara E: Id proteins in cell cycle control and cellular senescence. Oncogene. 20:8317–8325. 2001. View Article : Google Scholar : PubMed/NCBI
15	Benezra R, Rafii S and Lyden D: The Id proteins and angiogenesis. Oncogene. 20:8334–8341. 2001. View Article : Google Scholar : PubMed/NCBI
16	Ellmeier W, Aguzzi A, Kleiner E, Kurzbauer R and Weith A: Mutually exclusive expression of a helix-loop-helix gene and N-myc in human neuroblastomas and in normal development. EMBO J. 11:2563–2571. 1992.PubMed/NCBI
17	Israel MA, Hernandez MC, Florio M, Andres-Barquin PJ, Mantani A, Carter JH and Julin CM: Id gene expression as a key mediator of tumor cell biology. Cancer Res. 59:1726s–1730s. 1999.PubMed/NCBI
18	Lyden D, Young AZ, Zagzag D, Yan W, Gerald W, O’Reilly R, Bader BL, Hynes RO, Zhuang Y, Manova K and Benezra R: Id1 and Id3 are required for neurogenesis, angiogenesis and vascularization of tumour xenografts. Nature. 401:670–677. 1999. View Article : Google Scholar : PubMed/NCBI
19	Maruyama H, Kleeff J, Wildi S, Friess H, Büchler MW, Israel MA and Korc M: Id-1 and Id-2 are overexpressed in pancreatic cancer and in dysplastic lesions in chronic pancreatitis. AmJ Pathol. 155:815–822. 1999. View Article : Google Scholar : PubMed/NCBI
20	Lin CQ, Singh J, Murata K, Itahana Y, Parrinello S, Liang SH, Gillett CE, Campisi J and Desprez PY: A role for Id-1 in the aggressive phenotype and steroid hormone response of human breast cancer cells. Cancer Res. 60:1332–1340. 2000.PubMed/NCBI
21	Langlands K, Down GA and Kealey T: Id proteins are dynamically expressed in normal epidermis and dysregulated in squamous cell carcinoma. Cancer Res. 60:5929–5933. 2000.PubMed/NCBI
22	Wilson JW, Deed RW, Inoue T, Balzi M, Becciolini A, Faraoni P, Potten CS and Norton JD: Expression of Id helix-loop-helix proteins in colorectal adenocarcinoma correlates with p53 expression and mitotic index. Cancer Res. 61:8803–8810. 2001.PubMed/NCBI
23	Schindl M, Schoppmann SF, Ströbel T, Heinzl H, Leisser C, Horvat R and Birner P: Level of Id-1protein expression correlates with poor differentiation, enhanced malignant potential and more aggressive clinical behavior of epithelial ovarian tumors. Clin Cancer Res. 9:779–785. 2003.
24	Itahana Y, Singh J, Sumida T, Coppe JP, Parrinello S, Bennington JL and Desprez PY: Role of Id-2 in the maintenance of a differentiated and noninvasive phenotype in breast cancer cells. Cancer Res. 63:7098–7105. 2003.PubMed/NCBI
25	Coppe JP, Itahana Y, Moore DH, Bennington JL and Desprez PY: Id-1 and Id-2 proteins as molecular markers for human prostate cancer progression. Clin Cancer Res. 10:2044–2051. 2004. View Article : Google Scholar : PubMed/NCBI
26	Umetani N, Takeuchi H, Fujimoto A, Shinozaki M, Bilchik AJ and Hoon DS: Epigenetic inactivation of ID4 in colorectal carcinomas correlates with poor differentiation and unfavorable prognosis. Clin Cancer Res. 10:7475–7483. 2004. View Article : Google Scholar : PubMed/NCBI
27	de Candia P, Benera R and Solit DB: A role for Id proteins in mammary gland physiology and tumorigenesis. Adv Cancer Res. 92:81–94. 2004.PubMed/NCBI
28	Stighall M, Manetopoulos C, Axelson H and Landberg G: High ID2 protein expression correlates with a favourable prognosis in patients with primary breast cancer and reduces cellular invasiveness of breast cancer cells. Int J Cancer. 115:403–411. 2005. View Article : Google Scholar
29	Umetani N, Mori T, Koyanagi K, Shinozaki M, Kim J, Giuliano AE and Hoon DS: Aberrant hypermethylation of ID4 gene promoter region increases risk of lymph node metastasis in T1 breast cancer. Oncogene. 24:4721–4727. 2005. View Article : Google Scholar : PubMed/NCBI
30	Matsuda Y, Yamagiwa S, Takamura M, Honda Y, Ishimoto Y, Ichida T and Aoyagi Y: Overexpressed Id-1 is associated with a high risk of hepatocellular carcinoma development in patients with cirrhosis without transcriptional repression of p16. Cancer. 104:1037–1044. 2005. View Article : Google Scholar : PubMed/NCBI
31	Lee TK, Poon RT, Yuen AP, Ling MT, Wang XH, Wong YC, Guan XY, Man K, Tang ZY and Fan ST: Regulation of angiogenesis by Id-1 through hypoxia-inducible factor-1a-mediated vascular endothelial growth factor up-regulation in hepatocellular carcinoma. Clin Cancer Res. 12:6910–6919. 2006. View Article : Google Scholar : PubMed/NCBI
32	Drummond DC, Noble CO, Kirpotin DB, Guo Z, Scott GK and Benz CC: Clinical development of histone deacetylase inhibitors as anticancer agents. Annu Rev Pharmacol Toxicol. 45:495–528. 2005. View Article : Google Scholar : PubMed/NCBI
33	Liu T, Kuljaca S, Tee A and Marshall GM: Histone deacetylase inhibitors: multifunctional anticancer agents. Cancer Treat Rev. 32:157–165. 2006. View Article : Google Scholar : PubMed/NCBI
34	Khan O and La Thangue NB: HDAC inhibitors in cancer biology: Emerging mechanisms and clinical applications. Immunol Cell Biol. 90:85–94. 2012. View Article : Google Scholar : PubMed/NCBI
35	Rosato RR, Almenara JA, Dai Y and Grant S: Simultaneous activation of the intrinsic and extrinsic pathways by histone deacetylase (HDAC) inhibitors and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) synergistically induces mitochondrial damage and apoptosis in human leukemia cells. Mol Cancer Ther. 2:1273–1284. 2003.
36	Emanuele S, Lauricella M and Tesoriere G: Histone deacetylase inhibitors: Apoptotic effects and clinical implications. Int J Oncol. 33:637–646. 2008.PubMed/NCBI
37	Zhang J, Kan S, Huang B, Hao Z, Mak TW and Zhong Q: Mule determines the apoptotic response to HDAC inhibitors by targeted ubiquitination and destruction of HDAC2. Genes Dev. 25:2610–2618. 2011. View Article : Google Scholar : PubMed/NCBI
38	Vigushin DM and Coombes RC: Histone deacetylase inhibitors in cancer treatment. Anticancer Drugs. 13:1–13. 2002. View Article : Google Scholar
39	Lin HY and Chen CS, Lin SP, Weng JR and Chen CS: Targeting histone deacetylase in cancer therapy. Med Res Rev. 26:397–413. 2006. View Article : Google Scholar : PubMed/NCBI
40	Xu WS, Parmigiani RB and Marks PA: Histone deacetylase inhibitors: molecular mechanisms of action. Oncogene. 26:5541–5552. 2007. View Article : Google Scholar : PubMed/NCBI
41	Tsunedomi R, Ogawa Y, Iizuka N, Sakamoto K, Tamesa T, Moribe T and Oka M: The assessment of methylated BASP1 and SRD5A2 levels in the detection of early hepatocellular carcinoma. Int J Oncol. 36:205–212. 2010.PubMed/NCBI
42	Kouzarides T: Histone acetylases and deacetylases in cell proliferation. Curr Opin Genet Dev. 9:40–48. 1999. View Article : Google Scholar
43	Marks PA, Richon VM and Rifkind RA: Histone deacetylase inhibitors:inducers of differentiation or apotosis of transformed cells. J Natl Cancer Inst. 92:1210–1216. 2000. View Article : Google Scholar : PubMed/NCBI
44	Timmermann S, Lehrmann H, Polesskaya A and Harel-Bellan A: Histone acetylation and disease. Cell Mol Life Sci. 58:728–736. 2001. View Article : Google Scholar : PubMed/NCBI
45	Ogawa K, Yasumura S, Atarashi Y, Minemura M, Miyazaki T, Iwamoto M, Higuchi K and Watanabe A: Sodium butyrate enhances Fas-mediated apoptosis of human hepatoma cells. J Hepatol. 40:278–284. 2004. View Article : Google Scholar : PubMed/NCBI
46	Joseph J, Mudduluru G, Antony S, Vashistha S, Ajitkumar P and Somasundaram K: Expression profiling of sodium butyrate (NaB)-treated cells: identification of regulation of genes related to cytokine signaling and cancer metastasis by NaB. Oncogene. 23:6304–6315. 2004. View Article : Google Scholar
47	Chiba T, Yokosuka O, Arai M, Tada M, Fukai K, Imazeki F, Kato M, Seki N and Saisho H: Identification of genes up-regulated by histone deacetylase inhibition with cDNA microarray and exploration of epigenetic alterations on hepatoma cells. J Hepatol. 41:436–445. 2004. View Article : Google Scholar : PubMed/NCBI
48	Chen J, Saha P, Kornbluth S, Dynlacht BD and Dutta A: Cyclin-binding motifs are essential for the function of p21CIP1. Mol Cell Biol. 16:4673–4682. 1996.PubMed/NCBI
49	Shin JY, Kim HS, Park J, Park JB and Lee JY: Mechanism for inactivation of the KIP family cyclin-dependent kinase inhibitor genes in gastric cancer cells. Cancer Res. 60:262–265. 2000.PubMed/NCBI
50	Han JW, Ahn SH, Kim YK, Bae GU, Yoon JW, Hong S, Lee HY, Lee YW and Lee HW: Activation of p21^WAF1/Cip1 transcription through Sp1 sites by histone deacetylase inhibitor apicidin. J Biol Chem. 276:42084–42090. 2001.PubMed/NCBI