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Cellular and Molecular Biology

MET exon 14 skipping mutation drives cancer progression and recurrence via activation of SMAD2 signalling

British Journal of Cancer volume 130, pages 380–393 (2024)Cite this article

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Abstract

Background

c-Met encoded by the proto-oncogene MET, also known as hepatocyte growth factor (HGF) receptor, plays a crucial role in cellular processes. MET exon 14 skipping alteration (METΔ14EX) is a newly discovered MET mutation. SMAD2 is an important downstream transcription factor in TGF-β pathway. Unfortunately, the mechanisms by which METΔ14EX leads to oncogenic transformation are scarcely understood. The relationship between METΔ14EX and SMAD2 has not been studied yet.

Methods

We generate METΔ14EX models by CRISPR-Cas9. In vitro transwell, wound-healing, soft-agar assay, in vivo metastasis and subcutaneous recurrence assay were used to study the role of METΔ14EX in tumour progression. RNA-seq, Western blotting, co-immunoprecipitation (CO-IP) and immunofluorescent were performed to explore the interaction between c-Met and SMAD2.

Results

Our results demonstrated that METΔ14EX, independent of HGF, can prolong the constitutive activation of c-Met downstream signalling pathways by impeding c-Met degradation and facilitating tumour metastasis and recurrence. Meanwhile, METΔ14EX strengthens the interaction between c-Met and SMAD2, promoting SMAD2 phosphorylation. Therapeutically, MET inhibitor crizotinib impedes METΔ14EX-mediated tumour metastasis by decreasing SMAD2 phosphorylation.

Conclusions

These data elucidated the previously unrecognised role of METΔ14EX in cancer progression via activation of SMAD2 independent of TGF-β, which helps to develop more effective therapies for such patients.

METΔ14EX alteration significantly triggers tumour progression via activation of SMAD2 signalling that are involved in activating tumour invasion, metastasis and recurrence. On the left, in the MET wild-type (METWT), the juxtamembrane (JM) domain is involved in the regulation of tyrosine kinase activity, receptor degradation, and caspase cleavage. On the right, the METΔ14EX mutation leads to the loss of the juxtamembrane domain, resulting in an abnormal MET protein lacking a CBL-binding site. This causes the accumulation of truncated MET receptors followed by constitutive activation of the MET signalling pathway. Thus, the METΔ14EX-mutated protein has strong binding and phosphorylation to SMAD2, which results in the phosphorylation of a large number of SMAD2/3 proteins that combine with SMAD4 to form a complex in the nucleus, activating downstream signalling pathways, such as EMT and ECM remodelling, resulting in tumour progression and recurrence. TF transcription factor.

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Fig. 1: METΔ14EX alteration promotes cellular migration and invasion in vitro, independent of HGF.
Fig. 2: METΔ14EX alteration enhances tumour progression, metastasis and recurrence in vivo.
Fig. 3: RNA-seq analysis reveals that METΔ14EX alteration remarkably upregulated dominant cell movement-associated gene signatures.
Fig. 4: METΔ14EX alteration activates SMAD2 signalling via promoting phosphorylation of SMAD2.
Fig. 5: METΔ14EX-mediated tumour metastasis through activation of SMAD2 signalling.
Fig. 6: Crizotinib dramatically represses METΔ14EX-mediated tumour growth as well as metastasis.

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Data availability

All data generated or analysed during this study are included in this published article and its supplementary information files.

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Funding

This study was supported by the National Natural Science Foundation of China (NSFC 82273293) and Shanghai Municipal Health Commission Health Industry Clinical Research Project (20224Y0120).

Author information

Authors and Affiliations

  1. Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China

    Qiaoyan Liang, Yajun Hu, Qingyun Yuan, Min Yu & Bing Zhao

  2. Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China

    Qiaoyan Liang, Yajun Hu, Qingyun Yuan, Min Yu & Bing Zhao

  3. Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China

    Huijie Wang

  4. Department of Medical Oncology, Shanghai Cancer Center, Fudan University, Shanghai, China

    Huijie Wang

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  1. Qiaoyan Liang
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Contributions

BZ, MY, HW and QL performed all of the experiments. YH and QY participated in the research. QL, BZ and MY designed experiments, analysed data, and wrote the manuscript. HW provided c-Met inhibitors. QL and BZ are the guarantors of this work and, as such, had full access to all the data in the study and took responsibility for the integrity of the data and the accuracy of the data analysis. All authors read and approved the final manuscript.

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Correspondence to Min Yu, Huijie Wang or Bing Zhao.

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Liang, Q., Hu, Y., Yuan, Q. et al. MET exon 14 skipping mutation drives cancer progression and recurrence via activation of SMAD2 signalling. Br J Cancer 130, 380–393 (2024). https://doi.org/10.1038/s41416-023-02495-5

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