Identification of novel molecular targets regulated by tumor suppressive miR-375 induced by histone acetylation in esophageal squamous cell carcinoma

Isozaki,Yuka; Hoshino,Isamu; Nohata,Nijiro; Kinoshita,Takashi; Akutsu,Yasunori; Hanari,Naoyuki; Mori,Mikito; Yoneyama,Yasuo; Akanuma,Naoki; Takeshita,Nobuyoshi; Maruyama,Tetsuro; Seki,Naohiko; Nishino,Norikazu; Yoshida,Minoru; Matsubara,Hisahiro

doi:10.3892/ijo.2012.1537

Identification of novel molecular targets regulated by tumor suppressive miR-375 induced by histone acetylation in esophageal squamous cell carcinoma

Authors:
- Yuka Isozaki
- Isamu Hoshino
- Nijiro Nohata
- Takashi Kinoshita
- Yasunori Akutsu
- Naoyuki Hanari
- Mikito Mori
- Yasuo Yoneyama
- Naoki Akanuma
- Nobuyoshi Takeshita
- Tetsuro Maruyama
- Naohiko Seki
- Norikazu Nishino
- Minoru Yoshida
- Hisahiro Matsubara
View Affiliations

Affiliations: Department of Frontier Surgery, Chiba University Graduate School of Medicine, Chiba, Japan, Department of Functional Genomics, Chiba University Graduate School of Medicine, Chiba, Japan, Graduate School of Life Sciences and Systems Engineering, Kyushu Institute of Technology, Wakamatsu-ku, Kitakyushu, Japan, Chemical Genetic Laboratory, RIKEN, Wako, Saitama, Japan
Published online on: June 28, 2012 https://doi.org/10.3892/ijo.2012.1537
Pages: 985-994

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

The aim of this study was to determine whether histone acetylation regulates tumor suppressive microRNAs (miRNAs) in esophageal squamous cell carcinoma (ESCC) and to identify genes which are regulated by these miRNAs. We identified a miRNA that was highly upregulated in an ESCC cell line by cyclic hydroxamic acid-containing peptide 31 (CHAP31), one of the histone deacetylase inhibitors (HDACIs), using a miRNA array analysis. miR-375 was strongly upregulated by CHAP31 treatment in an ESCC cell line. The expression levels of the most upregulated miRNA, miR-375 were analyzed by quantitative real-time PCR in human ESCC specimens. The tumor suppressive function of miR-375 was revealed by restoration of miR-375 in ESCC cell lines. We performed a microarray analysis to identify target genes of miR-375. The mRNA and protein expression levels of these genes were verified in ESCC clinical specimens. LDHB and AEG-1/MTDH were detected as miR‑375-targeted genes. The restoration of miR-375 suppressed the expression of LDHB and AEG-1/MTDH. The ESCC clinical specimens exhibited a high level of LDHB expression at both the mRNA and protein levels. A loss-of-function assay using a siRNA analysis was performed to examine the oncogenic function of the gene. Knockdown of LDHB by RNAi showed a tumor suppressive function in the ESCC cells. The correlation between gene expression and clinicopathological features was investigated by immunohistochemistry for 94 cases of ESCC. The positive staining of LDHB correlated significantly with lymph node metastasis and tumor stage. It also had a tendency to be associated with a poor prognosis. Our results indicate that HDACIs upregulate miRNAs, at least some of which act as tumor suppressors. LDHB, which is regulated by the tumor suppressive miR-375, may therefore act as an oncogene in ESCC.

View Figures

View References

1.	Enzinger PC and Mayer RJ: Esophageal cancer. N Engl J Med. 349:2241–2252. 2003. View Article : Google Scholar : PubMed/NCBI
2.	Akutsu Y and Matsubara H: The significance of lymph node status as a prognostic factor for esophageal cancer. Surg Today. 41:1190–1195. 2011. View Article : Google Scholar : PubMed/NCBI
3.	National Cancer Institute (Bethesda, MD, USA). The Surveillance, Epidemiology and End Results (SEER) Program. Cancer Statistics Review. 2007.
4.	Boumber Y and Issa JP: Epigenetics in cancer: what’s the future? Oncology (Williston Park). 25:220–226. 2282011.
5.	Hoshino I and Matsubara H: Recent advances in histone deacetylase targeted cancer therapy. Surg Today. 40:809–815. 2010. View Article : Google Scholar : PubMed/NCBI
6.	Hoshino I, Matsubara H, Ochiai T, et al: Histone deacetylase inhibitor FK228 activates tumor suppressor Prdx1 with apoptosis induction in esophageal cancer cells. Clin Cancer Res. 11:7945–7952. 2005. View Article : Google Scholar : PubMed/NCBI
7.	Murakami K, Matsubara H, Hoshino I, et al: CHAP31 induces apoptosis only via the intrinsic pathway in human esophageal cancer cells. Oncology. 78:62–74. 2010. View Article : Google Scholar : PubMed/NCBI
8.	Hoshino I, Matsubara H, Ochiai T, et al: Gene expression profiling induced by histone deacetylase inhibitor, FK228, in human esophageal squamous cancer cells. Oncol Rep. 18:85–92. 2007.PubMed/NCBI
9.	Bartel DP: MicroRNAs: genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004. View Article : Google Scholar : PubMed/NCBI
10.	Kloosterman WP and Plasterk RH: The diverse functions of microRNAs in animal development and disease. Dev Cell. 11:441–450. 2006. View Article : Google Scholar : PubMed/NCBI
11.	Lu J, Getz G, Miska EA, et al: MicroRNA expression profiles classify human cancers. Nature. 435:834–838. 2005. View Article : Google Scholar : PubMed/NCBI
12.	Calin GA and Croce CM: MicroRNA signatures in human cancers. Nat Rev Cancer. 6:857–866. 2006. View Article : Google Scholar : PubMed/NCBI
13.	Chiyomaru T, Enokida H, Nakagawa M, et al: miR-145 and miR-133a function as tumour suppressors and directly regulate FSCN1 expression in bladder cancer. Br J Cancer. 102:883–891. 2010. View Article : Google Scholar : PubMed/NCBI
14.	Ichimi T, Enokida H, Seki N, et al: Identification of novel microRNA targets based on microRNA signatures in bladder cancer. Int J Cancer. 125:345–352. 2009. View Article : Google Scholar : PubMed/NCBI
15.	Nohata N, Hanazawa T, Seki N, et al: Tumor suppressive microRNA-375 regulates oncogene AEG-1/MTDH in head and neck squamous cell carcinoma (HNSCC). J Hum Genet. 56:595–601. 2011. View Article : Google Scholar : PubMed/NCBI
16.	Sugimoto T, Seki N, Hata A, et al: The galanin signaling cascade is a candidate pathway regulating oncogenesis in human squamous cell carcinoma. Genes Chromosomes Cancer. 48:132–142. 2009. View Article : Google Scholar : PubMed/NCBI
17.	Wagner JM, Hackanson B, Lübbert M, et al: Histone deacetylase (HDAC) inhibitors in recent clinical trials for cancer therapy. Clin Epigenetics. 1:117–136. 2010. View Article : Google Scholar : PubMed/NCBI
18.	Ma X, Ezzeldin HH and Diasio RB: Histone deacetylase inhibitors: current status and overview of recent clinical trials. Drugs. 69:1911–1934. 2009. View Article : Google Scholar : PubMed/NCBI
19.	Kano M, Seki N, Matsubara H, et al: miR-145, miR-133a and miR-133b: Tumor-suppressive miRNAs target FSCN1 in esophageal squamous cell carcinoma. Int J Cancer. 127:2804–2814. 2010. View Article : Google Scholar : PubMed/NCBI
20.	Kikkawa N, Hanazawa T, Seki N, et al: miR-489 is a tumour-suppressive miRNA target PTPN11 in hypopharyngeal squamous cell carcinoma (HSCC). Br J Cancer. 103:877–884. 2010. View Article : Google Scholar : PubMed/NCBI
21.	Yoshino H, Chiyomaru T, Nakagawa M, et al: The tumour-suppressive function of miR-1 and miR-133a targeting TAGLN2 in bladder cancer. Br J Cancer. 104:808–818. 2011. View Article : Google Scholar : PubMed/NCBI
22.	Kinoshita T, Nohata N, Yoshino H, et al: Tumor suppressive microRNA-375 regulates lactate dehydrogenase B in maxillary sinus squamous cell carcinoma. Int J Oncol. 40:185–193. 2012.PubMed/NCBI
23.	Li X, Lin R and Li J: Epigenetic silencing of microRNA-375 regulates PDK1 expression in esophageal cancer. Dig Dis Sci. 56:2849–2856. 2011. View Article : Google Scholar : PubMed/NCBI
24.	Tsukamoto Y, Nakada C, Moriyama M, et al: MicroRNA-375 is downregulated in gastric carcinomas and regulates cell survival by targeting PDK1 and 14-3-3zeta. Cancer Res. 70:2339–2349. 2010. View Article : Google Scholar : PubMed/NCBI
25.	Kong KL, Kwong DL, Guan XY, et al: MicroRNA-375 inhibits tumour growth and metastasis in oesophageal squamous cell carcinoma through repressing insulin-like growth factor 1 receptor. Gut. 61:33–42. 2012. View Article : Google Scholar : PubMed/NCBI
26.	Rountree MR, Bachman KE, Baylin SB, et al: DNA methylation, chromatin inheritance, and cancer. Oncogene. 20:3156–3165. 2001. View Article : Google Scholar : PubMed/NCBI
27.	Zha X, Wang F, Zhang H, et al: Lactate dehydrogenase B is critical for hyperactive mTOR-mediated tumorigenesis. Cancer Res. 71:13–18. 2011. View Article : Google Scholar : PubMed/NCBI
28.	Chen Y, Zhang H, Xiao X, et al: Elevation of serum l-lactate dehydrogenase B correlated with the clinical stage of lung cancer. Lung Cancer. 54:95–102. 2006. View Article : Google Scholar : PubMed/NCBI
29.	Hu G, Wei Y and Kang Y: The multifaceted role of MTDH/AEG-1 in cancer progression. Clin Cancer Res. 15:5615–5620. 2009. View Article : Google Scholar : PubMed/NCBI
30.	Yu C, Chen K, Song L, et al: Overexpression of astrocyte elevated gene-1 (AEG-1) is associated with esophageal squamous cell carcinoma (ESCC) progression and pathogenesis. Carcinogenesis. 30:894–901. 2009. View Article : Google Scholar : PubMed/NCBI