Gastric Cancer

, Volume 20, Issue 6, pp 960–969 | Cite as

Expression and function of Uc.160+, a transcribed ultraconserved region, in gastric cancer

  • Ririno Honma
  • Keisuke Goto
  • Naoya Sakamoto
  • Yohei Sekino
  • Kazuhiro Sentani
  • Naohide Oue
  • Wataru Yasui
Original Article
  • 223 Downloads

Abstract

Background

Transcribed ultraconserved regions (T-UCRs) are a novel class of noncoding RNAs that are highly conserved among the orthologous regions in most vertebrates. It has been reported that T-UCRs have distinct signatures in human cancers. We previously discovered the downregulation of T-UCR expression in gastric cancer (GC), indicating that T-UCRs could play an important role in GC biology. Uc.160+, a T-UCR reported to be downregulated in human cancer, has not been examined in GC.

Methods

We analyzed the expression pattern of Uc.160+ in nonneoplastic and tumor tissues of the stomach by using uantitative reverse transcription polymerase chain reaction (qRT-PCR) and in situ hybridization (ISH), specifically focusing on the mechanism of transcriptional regulation and target genes that are regulated by T-UCRs. We also attempted to determine the effect of Uc.160+ expression on biological features of GC cell lines by Western blotting.

Results

On the basis of the qRT-PCR and ISH results, Uc.160+ expression in adenoma and GC tissues was clearly downregulated compared with that in nonneoplastic mucosa tissues of the stomach. Cancer-specific DNA methylation in the promoter region of Uc.160 was observed by bisulfite genomic DNA sequencing analysis. The effect of DNA methylation on Uc.160+ expression was further confirmed by reporter gene assay. We also revealed that Uc.160+ inhibited the phosphorylation of Akt by regulating phosphatase and tensin homolog (PTEN) expression.

Conclusions

These results indicate that Uc.160+ could possibly have a tumor suppressive role in GC.

Keywords

Gastric cancer Noncoding RNA Transcribed ultraconserved region DNA methylation 

Notes

Acknowledgements

We thank Shinichi Norimura for excellent technical assistance. We thank the Analysis Center of Life Science, Hiroshima University, for the use of its facilities. This work was supported by Grants-in-Aid for Scientific Research (JP15H04713, JP16K08691) and Grants-in-Aid for Challenging Exploratory Research (26670175, JP16K15247) from the Japan Society for the Promotion of Science.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Human rights statement and informed consent

All procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation (institutional and national) and with the Helsinki Declaration of 1964 and later versions. Informed consent or substitute for it was obtained from all patients for their being included in the study.

Supplementary material

10120_2017_714_MOESM1_ESM.tif (562 kb)
Fig. S1 Functional analysis using Uc.160+-silenced cancer cells. a Uc.160+ expression in normal colon and colon cancer cells. b Uc.160+ expression in DLD-1 cells transfected with negative control or two different small interfering RNAs. c The growth-promotive effect of Uc.160+ knockout in DLD-1 cells. The results are expressed as the mean ± standard deviation of triplicate measurements (TIFF 562 kb).
10120_2017_714_MOESM2_ESM.xlsx (40 kb)
Table S1 A 260/A 280 ratios (XLSX 40 kb).

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Copyright information

© The International Gastric Cancer Association and The Japanese Gastric Cancer Association 2017

Authors and Affiliations

  • Ririno Honma
    • 1
  • Keisuke Goto
    • 2
  • Naoya Sakamoto
    • 1
  • Yohei Sekino
    • 1
  • Kazuhiro Sentani
    • 1
  • Naohide Oue
    • 1
  • Wataru Yasui
    • 1
  1. 1.Department of Molecular PathologyHiroshima University Institute of Biomedical and Health SciencesHiroshimaJapan
  2. 2.Cancer Biology ProgramUniversity of Hawaii Cancer CenterHonoluluUSA

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