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DLX6-AS1/miR-204-5p/OCT1 positive feedback loop promotes tumor progression and epithelial–mesenchymal transition in gastric cancer

  • Yu Liang
  • Chun-Dong Zhang
  • Cheng Zhang
  • Dong-Qiu DaiEmail author
Original Article

Abstract

Background

Accumulating evidence indicates that long non-coding RNAs (lncRNAs) participate in progression of gastric cancer (GC). Nevertheless, the function and expression level of DLX6-AS1 in GC remain unknown.

Methods

We explored the sequencing data of DLX6-AS1 downloaded from The Cancer Genome Atlas. The expression of DLX6-AS1, miR-204-5p and OCT1 in 56 GC patients and GC cell lines was quantified by qRT-PCR and western blotting. Furthermore, we performed in vitro functional assays to assess proliferation, invasion and migration of GC cells by knockdown of DLX6-AS1. The expression level of epithelial–mesenchymal transition (EMT)-related genes was also determined by qRT-PCR and western blotting. Actin remodeling was detected by F-actin phalloidin staining. The luciferase reporter assay and chromatin immunoprecipitation assay was utilized to confirm the bioinformatic prediction. The function of the DLX6-AS1/miR-204-5p/OCT1 axis in GC proliferation was clarified by rescue assays.

Results

We first demonstrated that DLX6-AS1 was upregulated in GC tissues and cell lines and was associated with T3/T4 invasion, distant metastasis and poor clinical prognosis. Further functional analysis showed that downregulation of DLX6-AS1 inhibited GC cell proliferation, migration, invasion and EMT in vitro. Mechanistic investigation indicated that DLX6-AS1 acted as a cancer-promoting competing endogenous RNA (ceRNA) by binding miR-204-5p and upregulating OCT1. Moreover, the transcription factor OCT1 was confirmed to enhance DLX6-AS1 expression by targeting the promoter region.

Conclusions

This study revealed that OCT1-induced DLX6-AS1 promoted GC progression and the EMT via the miR-204-5p/OCT1 axis, suggesting that this lncRNA might be a promising prognostic biomarker and therapeutic target for GC.

Keywords

DLX6-AS1 Gastric cancer miR-204-5p OCT1 EMT 

Notes

Funding

The study was funded by the National Natural Science Foundation of China (Grant number 30572162); the Natural Science Foundation of Liaoning Province (Grant number 201602817); Chun-Dong Zhang was supported in part by the Japan China Sasakawa Medical Fellowship (Grant number 2017816).

Compliance with ethical standards

Conflict of interest

Yu Liang declares that he has no conflict of interest. Chun-Dong Zhang declares that he has no conflict of interest. Cheng Zhang declares that he has no conflict of interest. Dong-Qiu Dai declares that he has no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study.

Supplementary material

10120_2019_1002_MOESM1_ESM.pdf (18.2 mb)
Supplementary file1 (PDF 18683 kb)

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Copyright information

© The International Gastric Cancer Association and The Japanese Gastric Cancer Association 2019

Authors and Affiliations

  1. 1.Department of Gastrointestinal SurgeryThe Fourth Affiliated Hospital of China Medical UniversityShenyangChina
  2. 2.Department of Gastrointestinal Surgery, Graduate School of MedicineUniversity of TokyoTokyoJapan
  3. 3.Division of EpigenomicsNational Cancer Center Research InstituteTokyoJapan
  4. 4.Cancer CenterThe Fourth Affiliated Hospital of China Medical UniversityShenyangChina

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