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Kindlin-2–miR-1258–TCF4 feedback loop promotes hepatocellular carcinoma invasion and metastasis

  • Original Article—Liver, Pancreas, and Biliary Tract
  • Published:
Journal of Gastroenterology Aims and scope Submit manuscript

Abstract

Background

Upregulated Kindlin-2 expression in hepatocellular carcinoma (HCC) correlates with metastasis and poor prognosis. In this study, we investigated the molecular mechanism of Kindlin-2 in HCC.

Methods

Kindlin-2 downstream pathways were explored through microRNA sequencing. The Kindlin-2–miR-1258–TCF4 axis was verified using bisulfite sequencing, a luciferase reporter assay, quantitative real-time PCR, and rescue assays. Binding of TCF4 to the Kindlin-2 promoter was confirmed by promoter activity analysis and chromatin immunoprecipitation.

Results

MiRNA sequencing identified miR-1258 as a downstream effector of Kindlin-2. MiR-1258 expression was increased following Kindlin-2 knockdown and decreased after Kindlin-2 overexpression. Next, we identified transcription factor 7 like 2 (TCF7L2 or TCF4) as a target of miR-1258 and found that Kindlin-2 upregulated TCF4 expression by epigenetically suppressing miR-1258 in HCC. Furthermore, our results suggest that TCF4 binds to the Kindlin-2 promotor to enhance its transcription. Therefore, Kindlin-2–miR-1258–TCF4 interaction creates a positive feedback loop. Functional assays and animal experiments demonstrated critical roles of miR-1258 and TCF4 in HCC cell migration in vitro and HCC metastasis in vivo. In HCC tissues, Kindlin-2 expression correlated negatively with miR-1258 expression and positively with TCF4 expression. Meanwhile, miR-1258 expression correlated negatively with TCF4 expression.

Conclusions

This study illustrates a novel integrin-independent signaling pathway, Kindlin-2–miR-1258–TCF4, that regulates HCC invasion and metastasis and identifies Kindlin-2 as a promising therapeutic target in HCC.

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Abbreviations

HCC:

Hepatocellular carcinoma

EMT:

Epithelial–mesenchymal transition

miR:

MicroRNA

SDS-PAGE:

Sodium dodecyl sulfate–polyacrylamide gel electrophoresis

shRNA:

Short hairpin RNAs

CPM:

Counts per million reads

qRT-PCR:

Quantitative real-time PCR

CRISPR:

Clustered regularly interspaced short palindromic repeats

Cas 9:

CRISPR-associated protein 9

ChIP:

Chromatin immunoprecipitation

DNMT:

DNA methyltransferase

5-aza-CdR:

5-Aza-2’-deoxycytidine

TCF4:

Transcription factor 7 like 2

FERM:

4.1-Ezrin-ridixin-moesin

3′-UTR:

3′-Untranslated region

CDS:

Coding sequence

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Acknowledgements

This work was supported by grants from the National Natural Science Foundation of China (Grant number: 81802882), Joint Funds for the innovation of Science and Technology, Fujian province (Grant number: 2018Y9104, 2018Y9108), and the Natural Science Foundation of Fujian Province (Grant number: 2018J01251).

Funding

National Natural Science Foundation of China, 81802882, Jie Lin, Joint Funds for the innovation of Science and Technology, Fujian province, 2018Y9104, Wansong Lin, 2018Y9108, Yunbin Ye, Natural Science Foundation of Fujian Province, 2018J01251, Jie Lin.

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Author notes

  1. Wansong Lin and Jie Lin have contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Immuno-Oncology, Fujian Medical University Cancer Hospital, Fujian Cancer Hospital, No.420, Fuma Road, Jinan District, Fuzhou, 350014, Fujian, China

    Wansong Lin, Jieyu Li, Shuping Chen & Yunbin Ye

  2. Fujian Key Laboratory of Translational Cancer Medicine, No.420, Fuma Road, Jinan District, Fuzhou, 350014, Fujian, China

    Wansong Lin, Jieyu Li, Shuping Chen & Yunbin Ye

  3. Department of Pathology, Shengli Clinical Medical College of Fujian Medical University, Fujian Provincial Hospital, No.134, Dong Street, Gulou District, Fuzhou, 350001, Fujian, China

    Jie Lin, Ying Lin, Yijuan Wu, Lingfeng Chen, Hui Cheng, Lin Liu & Xiaoyan Chen

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  1. Wansong Lin
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Contributions

LWS and LJ conceived and designed the study as well as performed the laboratory analysis. LY and WYJ performed the animal experiments and the ChIP analysis. LJY and CSP performed the western blotting analysis. CLF performed the qRT-PCR analysis in clinical samples. CH and LL collected the samples. LJ, LWS, CXY, and YYB contributed reagents, materials, and analysis tools. LWS and LJ wrote the manuscript. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Jie Lin or Yunbin Ye.

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Lin, W., Lin, J., Li, J. et al. Kindlin-2–miR-1258–TCF4 feedback loop promotes hepatocellular carcinoma invasion and metastasis. J Gastroenterol 57, 372–386 (2022). https://doi.org/10.1007/s00535-022-01866-8

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  • DOI: https://doi.org/10.1007/s00535-022-01866-8

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