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Coexpression of FOXK1 and vimentin promotes EMT, migration, and invasion in gastric cancer cells

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Abstract

In human gastric cancer (GC), the upregulation of FOXK1 and vimentin is frequently observed in cancer cells and correlates with increased malignancy. We report that FOXK1 synergizes with vimentin to promote GC invasion and metastasis via the induction of epithelial-mesenchymal transition (EMT). We showed that higher expression levels of FOXK1 were significantly associated with GC development. FOXK1 can physically interact with and stabilize vimentin. Moreover, a positive correlation between the expression of FOXK1 and vimentin was found in GC cells. Higher expression levels of these two proteins were significantly associated with differentiation, lymph node metastasis, AJCC stage, and poorer prognosis. Furthermore, the coexpression of FOXK1 and vimentin enhances cell metastasis through the induction of EMT in GC cells. However, the siRNA-mediated repression of vimentin in FOXK1-overexpressing cells reversed the EMT-like phenotype and reduced GC cell migration and invasion in vitro and in vivo. Altogether, our findings suggest that the vimentin-FOXK1 axis provides new insights into the molecular mechanisms underlying EMT regulation during GC progression and metastasis.

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Abbreviations

EMT:

Epithelial-to-mesenchymal transition

FOX:

The forkhead box

AJCC:

American Joint Committee on Cancer

GC:

Gastric cancer

TMA:

Tissue microarray

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Funding

This study was supported by grants from the National Natural Science Funds of China (81672875 and 81772964) and “President Foundation of Nanfang Hospital, Southern Medical University” (2012B009, 2013Z007) and high-level topic-matching funds of Nanfang Hospital (201347 and G201227). Guangdong Provincial Key Laboratory of Gastroenterology, Department of Gastroenterology, Nanfang Hospital, Southern Medical University; Guangzhou Pilot Project of Clinical and Translational Research Center (early gastrointestinal cancer, No. 7415696196402).

Author information

Authors and Affiliations

  1. Guangdong Provincial Key Laboratory of Gastroenterology, Department of Gastroenterology, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, China

    Hui Zhang, Xiaosheng Wu, Yizhi Xiao, Liqing Wu, Ying Peng, Weimei Tang, Guangnan Liu, Yong Sun, Jing Wang, Huiqiong Zhu, Mengwei Liu, Weiyu Dai, Ping Jiang, Aimin Li, Side Liu & Jide Wang

  2. Department of Gastroenterology, Hexian Memorial Affiliated Hospital of Southern Medical University, Guangzhou, 511400, China

    Hui Zhang

  3. Department of Medical Oncology, the First People’s Hospital of Yunnan Province, Kunming University of Science and Technology, Kunming, 650032, China

    Wenjing Zhang

  4. Department of General Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, China

    Guoxin Li

  5. Department of Gastroenterology, Longgang District People’s Hospital, Shenzhen, 518172, China

    Li Xiang, Side Liu & Jide Wang

Authors
  1. Hui Zhang
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  2. Xiaosheng Wu
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  18. Side Liu
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Corresponding authors

Correspondence to Li Xiang, Side Liu or Jide Wang.

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The authors declare that they have no conflict of interest.

Ethics approval

This study was conducted with the approval of the ethics committee of Southern Medical University.

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Article Summary

Our findings suggest that the vimentin-FOXK1 axis has provided new insights into the molecular mechanisms underlying EMT regulation during GC progression and metastasis.

Electronic supplementary material

Supplementary Fig. 1

Vimentin physically interacts with FOXK1 in SCG7901 cells. (A) Diagram showing the full-length and truncated (N terminus, FHA domain and C terminus) FOXK1 fragments. Immunoprecipitation was performed using cell lysates from SCG7901 cells transfected with the above four constructs. Anti-HA or normal mouse (nm) IgG was used as the bait to detect vimentin. The IP blot was probed with the indicated antibodies to show the input. (B) Full-length, N-terminal, IF rod domain or C-terminal vimentin protein fragments interacted with FOXK1. All images represent two or three independent experiments with identical results. (C) FOXK1 increases the stability of vimentin via a posttranslational mechanism. SCG7901 cells were transfected with vimentin. Twenty-four hours after transfection, cycloheximide (CHX) was added to the culture, and the levels of the FOXK1 protein at different time points were determined via western blotting. The half-life of FOXK1 increased from 3 to 7 h in the presence of vimentin. (PNG 63 kb)

High Resolution Image (TIFF 1036 kb)

Supplementary Fig. 2.

FOXK1 expression is positively correlated with vimentin expression in GC cells. These images are representative of 3 independent experiments with identical results. (PNG 242 kb)

High Resolution Image (TIFF 2193 kb)

Supplementary Fig. 3.

Assessment of migration and invasion. Cells stably transfected with vector or FOXK1 were transfected with vimentin siRNA 48 h later, and the migratory (A) and invasive (B) abilities of the cells were decreased. ** P < 0.05 and *** P < 0.01. The experiments were repeated at least three times. (PNG 651 kb)

High Resolution Image (TIFF 1667 kb)

Supplementary Fig. 4.

Gelatin zymography assay for MMP-2 and MMP-9 levels in SCG7901 cells. (A) Representative gelatin zymography results showing MMP2 and MMP9 activity. (B) Quantitative analysis of (A). The bars represent the intensity value of the bright band and were used for statistical analysis. N = 3, ****P < 0.001, vector vs. FOXK1 and FOXK1 vs. FOXK1-vimentin siRNA. (PNG 753 kb)

High Resolution Image (TIFF 481 kb)

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Zhang, H., Wu, X., Xiao, Y. et al. Coexpression of FOXK1 and vimentin promotes EMT, migration, and invasion in gastric cancer cells. J Mol Med 97, 163–176 (2019). https://doi.org/10.1007/s00109-018-1720-z

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  • DOI: https://doi.org/10.1007/s00109-018-1720-z

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