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Negative Regulation of Kruppel-Like Factor 4 on microRNA-106a at Upstream Transcriptional Level and the Role in Gastric Cancer Metastasis

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Abstract

Background

MicroRNAs are classes of endogenous noncoding RNAs that play a substantial role in tumor processes through regulating the targets at posttranscriptional level. However, little is known about the upstream transcription regulatory mechanism although it is a prerequisite for investigation of its aberrant expression and function.

Aims

This report evaluates miR-106a’s direct transcriptional factor from upstream level to in depth elucidate their communication in gastric cancer development.

Methods

Gastric cancer tissues were collected to analyze the miR-106a expression using real-time PCR methods. The combination of Kruppel (or Krüppel)-like factor 4 (KLF4) to miR-106a promoter was testified through bioinformatics followed by construction of luciferase reporter plasmid and chromatin immunoprecipitation assay. Functional experiments and mouse models for evaluating cell growth and metastasis were conducted to observe the biological effect of KLF4 on miR-106a. The interplay between KLF4 and miR-106a was tested with Wnt activator and confirmed in clinical specimens.

Results

The up-regulated miR-106a linked to gastric cancer metastasis and epithelial–mesenchymal transition. UCSC and JASPAR predicted the promoter sequence of miR-106a and its binding site with transcriptional factor KLF4. Construction of reporter gene further verified their direct combination at upstream level. Moreover, the inhibitory effect of KLF4 on the phenotype of gastric cancer cells could be restored by miR-106a. CHIR-induced experiment and clinical specimens confirmed the negative regulation of KLF4 on miR-106a.

Conclusions

Our findings provide novel direct insights into molecular mechanisms for interaction of KLF4 and miR-106a at upstream level and new ways for clinical application of KLF4–miR-106a axis in advanced gastric cancer metastasis.

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Abbreviations

KLF4:

Kruppel (or Krüppel)-like factor 4

EMT:

Epithelial–mesenchymal transition

ChIP:

Chromatin immunoprecipitation

TSS:

Transcription start site

H&E:

Hematoxylin and eosin

IHC:

Immunohistochemistry

qRT-PCR:

Quantitative real-time PCR

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Acknowledgments

This study was funded by China Postdoctoral Science Foundation (Grant Number 2018M633529), the Ningxia Natural Science Foundation (Grant Number NZ16148, NZ16276), Outstanding Young Teachers Development Foundation of Ningxia Higher Education Institutions (Grant Number NGY2016123) and Basic Research Project of Natural Science Foundation of Shaanxi Province (Grant Number 2017JM8120).

Author’s contribution

Meng Zhu and Shuixiang He conceived and supervised the study; Meng Zhu and Ning Zhang designed and performed experiments; Meng Zhu analyzed data and wrote the manuscript; Shuixiang He and Xinlan Lu made manuscript revisions.

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Authors and Affiliations

  1. Department of Gastroenterology, First Affiliated Hospital of Xi’an Jiaotong University, 277 West Yanta Road, Xi’an, 710061, Shaanxi, People’s Republic of China

    Meng Zhu, Xinlan Lu & Shuixiang He

  2. Department of Pathology, General Hospital of Ningxia Medical University, Yinchuan, Ningxia, People’s Republic of China

    Ning Zhang

Authors
  1. Meng Zhu
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  2. Ning Zhang
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  4. Shuixiang He
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Correspondence to Shuixiang He.

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Zhu, M., Zhang, N., Lu, X. et al. Negative Regulation of Kruppel-Like Factor 4 on microRNA-106a at Upstream Transcriptional Level and the Role in Gastric Cancer Metastasis. Dig Dis Sci 63, 2604–2616 (2018). https://doi.org/10.1007/s10620-018-5143-z

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  • DOI: https://doi.org/10.1007/s10620-018-5143-z

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