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Single-cell RNA sequencing reveals the lineage of malignant epithelial cells and upregulation of TAGLN2 promotes peritoneal metastasis in gastric cancer

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Abstract

Background

Peritoneal metastasis (PM) is an important factor contributing to poor prognosis in patients with gastric cancer (GC). Transcriptomic sequencing has been used to explore the molecular changes in metastatic cancers, but comparing the bulk RNA-sequencing data between primary tumors and metastases in PM studies is unreasonable due to the small proportion of tumor cells in PM tissues.

Methods

We performed single-cell RNA-sequencing analysis on four gastric adenocarcinoma specimens, including one primary tumor sample (PT), one adjacent nontumoral sample (PN), one peritoneal metastatic sample (MT) and one normal peritoneum sample (MN), from the same patient. Pseudotime trajectory analysis was used to display the process by which nonmalignant epithelial cells transform into tumor cells and then metastasize to the peritoneum. Finally, in vitro and in vivo assays were used to validate one of the selected genes that promote peritoneal metastasis.

Results

Single-cell RNA sequencing showed that a development curve was found from normal mucosa to tumor tissues and then into metastatic sites on peritoneum. TAGLN2 was found to trigger this metastasis process. The migration and invasion capability of GC cells were changed by downregulating and upregulating TAGLN2 expression. Mechanistically, TAGLN2 might modulate tumor metastasis via alterations in cell morphology and several signaling pathways, thus promoting epithelial–mesenchymal transition (EMT).

Conclusions

In summary, we identified and validated TAGLN2 as a novel gene involved in GC peritoneal metastasis. This study provided valuable insight into the mechanisms of GC metastasis and developed a potential therapeutic target to prevent GC cell dissemination.

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Data availability

The data that support the findings of this study are available on request from the corresponding author.

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Acknowledgements

This work was supported by grants awarded to Dr. Junjie Zhao (82002527), Prof. Xuefei Wang (81972228) and Prof. Yihong Sun (81872425) from National Natural Science Foundation of China.

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Author notes

  1. Chengbo Ji, Junjie Zhao and Hao chen contributed equally to this work.

Authors and Affiliations

  1. Department of General Surgery, Zhongshan Hospital, Fudan University, Shanghai, China

    Chengbo Ji, Junjie Zhao, Hao chen, Zhaoming Wang, Tianyi Cai, Chenyu Tian, Chen Wang, Dan Liu, Botian Ye, Min Fu, Jie Sun, Haojie Li, Xuefei Wang & Yihong Sun

  2. Gastric Cancer Center, Zhongshan Hospital, Fudan University, Shanghai, China

    Junjie Zhao, Hao chen, Zhaoming Wang, Tianyi Cai, Botian Ye, Min Fu, Jie Sun, Haojie Li, Xuefei Wang & Yihong Sun

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  1. Chengbo Ji
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Contributions

Conceptualization: CJ and JZ; methodology: CJ, HC, JZ. Data curation: CJ, CT, DL, CW; software and formal analysis: HC and CT; resources: TC, BY and MF; investigation: TC and JS; writing—original draft: CJ, JZ and ZW; writing—review and editing: XW and YS. Supervision, funding acquisition and project administration: HL and YS; all the authors read and approved the final manuscript.

Corresponding authors

Correspondence to Haojie Li or Yihong Sun.

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The authors declare no competing financial interests.

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All protocols in this study were approved by ethics committee of The Zhongshan Hospital and conducted in compliance with the Declaration of Helsinki.

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Single-cell RNA sequencing data were used from public GEO datasets. All the tissues from patients were obtained after informed consent.

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The original online version of this article was revised to correct Fig 4E caption and Figure 5.

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Ji, C., Zhao, J., chen, H. et al. Single-cell RNA sequencing reveals the lineage of malignant epithelial cells and upregulation of TAGLN2 promotes peritoneal metastasis in gastric cancer. Clin Transl Oncol 25, 3405–3419 (2023). https://doi.org/10.1007/s12094-023-03194-6

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