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GADD45G Interacts with E-cadherin to Suppress the Migration and Invasion of Esophageal Squamous Cell Carcinoma

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Abstract

Background/Aims

Esophageal squamous cell carcinoma (ESCC) is one of the most prevalent cancers with poor prognosis. Metastasis is the leading cause of cancer-related deaths. The growth arrest and DNA damage-inducible 45 gamma (GADD45G) has been reported to correlate with survival, invasion, and metastasis of ESCC. This study was aimed to investigate the role and mechanism of GADD45G in ESCC cell migration and invasion.

Methods

Both the effects of GADD45G and its need for E-cadherin to function on ESCC cell migration and invasion were determined through loss- and gain-of-function approaches via Transwell assays. The interaction between GADD45G and E-cadherin was detected by GST-pull down and IP assays. The expression of E-cadherin upon GADD45G overexpression was evaluated by RT-qPCR and western blot. The level of E-cadherin in cytoplasmic, nuclear, and membrane fractions was examined by western blot following subcellular fractionation.

Results

Knockdown of GADD45G increased the migration and invasion abilities of KYSE150 cells, while overexpression of GADD45G showed the opposite effects on YES2 and KYSE30 cells. GADD45G could interact with E-cadherin and enhanced its membrane level. Knockdown of E-cadherin abolished the inhibitory effects of GADD45G on ESCC cell migration and invasion. Intriguingly, dimer-dissociating mutant of GADD45G could not interact with E-cadherin and almost lost its ability to suppress the ESCC cell migration and invasion.

Conclusions

This study reveals a novel role for GADD45G in inhibiting the ESCC cell migration and invasion, which will provide a new insight in understanding the ESCC metastatic mechanism.

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Acknowledgments

We thank Wenzheng Zhang and Xuelian Zhao for construction the plasmids of pCS2 + MT and pGEX-6P-1 expressing wild-type GADD45G or its dimer-dissociating mutant A47R + I76E + L80E + A83R in our previous study.

Funding

This work was supported by the National Basic Research Program of China (973 Program) (2015CB553906 and 2015CB553904), the National Natural Science Foundation of China (81402283 and 81830086), the Education Department of Liaoning Province in China (Scientific Research Projects, L2016038) and the Natural Science Foundation of Liaoning (2015020304).

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Author notes

  1. Tongtong Li and Lele Xu have contributed equally to this work.

Authors and Affiliations

  1. Institute of Cancer Stem Cell, Dalian Medical University, No. 9 West Section, Lvshun South Road, Lvshunkou District, Dalian, 116044, China

    Tongtong Li, Lele Xu, Jinglei Teng, Yunping Ma, Wenzhong Liu, Qimin Zhan & Xuefeng Liu

  2. Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education/Beijing), Laboratory of Molecular Oncology, Peking University Cancer Hospital & Institute, No. 52 Fucheng Road, Haidian District, Beijing, 100142, China

    Yan Wang & Qimin Zhan

  3. Liaoning Key Laboratory of Proteomics, Dalian Medical University, No. 9 West Section, Lvshun South Road, Lvshunkou District, Dalian, 116044, China

    Xinming Chi & Shujuan Shao

  4. College of Stomatology, Dalian Medical University, No. 9 West Section, Lvshun South Road, Lvshunkou District, Dalian, 116044, China

    Yan Dong

  5. State Key Laboratory of Molecular Oncology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, No. 17 Panjiayuan Nanli, Chaoyang District, Beijing, 100021, China

    Qimin Zhan

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  1. Tongtong Li
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Correspondence to Xuefeng Liu.

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Li, T., Xu, L., Teng, J. et al. GADD45G Interacts with E-cadherin to Suppress the Migration and Invasion of Esophageal Squamous Cell Carcinoma. Dig Dis Sci 65, 1032–1041 (2020). https://doi.org/10.1007/s10620-019-05836-8

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