STAT3-EphA7 axis contributes to the progression of esophageal squamous cell carcinoma

Abstract

Background

Our previous study has revealed that EphA7 was upregulated in patient-derived esophageal squamous cell carcinoma (ESCC) xenografts with hyper-activated STAT3, but its mechanism was still unclear.

Materials and methods

To assess the association between EphA7 and STAT3, western blotting, immunofluorescence, ChIP assay, and qRT-PCR were conducted. Truncated mutation and luciferase assay were performed to examine the promoter activity of EphA7. CCK-8 assay and colony formation were performed to assess the proliferation of ESCC. Cell-derived xenograft models were established to evaluate the effects of EphA7 on ESCC tumor growth. RNA-seq analyses were used to assess the effects of EphA7 on related signals.

Results

In this study, EphA7 was found upregulated in ESCC cell lines with high STAT3 activation, and immunofluorescence also showed that EphA7 was co-localized with phospho-STAT3 in ESCC cells. Interestingly, suppressing STAT3 activation by the STAT3 inhibitor Stattic markedly inhibited the protein expression of EphA7 in ESCC cells, in contrast, activation of STAT3 by IL-6 obviously upregulated the protein expression of EphA7. Moreover, the transcription of EphA7 was also mediated by the activation of STAT3 in ESCC cells, and the −2000∼−1500 region was identified as the key promoter of EphA7. Our results also indicated that EphA7 enhanced the cell proliferation of ESCC, and silence of EphA7 significantly suppressed ESCC tumor growth. Moreover, EphA7 silence markedly abolished STAT3 activation-derived cell proliferation of ESCC. Additionally, RNA-seq analyses indicated that several tumor-related signaling pathways were significantly changed after EphA7 downregulation in ESCC cells.

Conclusion

Our results showed that the transcriptional expression of EphA7 was increased by activated STAT3, and the STAT3 signaling may act through EphA7 to promote the development of ESCC.

Keywords:

• STAT3
• EphA7
• transcription
• ESCC
• promoter

Ethical approval

This study was approved by the Review Board and Ethical Committee of the First Affiliated Hospital of Zhengzhou University.

Author contributions

Li Wang: Methodology, Investigation, Data curation, Writing, Original draft preparation, Visualization. Qiao-Feng Zhao: Methodology, Investigation. Bing-Bing Yang: Methodology, Investigation. Hui-Jie Liang: Validation, Investigation. Xian-E Zhang: Validation, Investigation. Xiao-Yan Zhang: Validation, Investigation. Wan-Jing Yang: Formal analysis. Zhi-Yu Guo: Software. Xin Xu: Writing, Reviewing and Editing. Fang Tian: Conceptualization, Writing, Reviewing and Editing, Project administration, Funding acquisition. Qing-Hua Wu: Conceptualization, Supervision, Funding acquisition. All authors contributed to and approved the final manuscript.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Data availability statement

All data generated or analyzed during this study are included in this published article. Data will be made available on request.

Additional information

Funding

This work was supported by the National Natural Sciences Foundation of China [U1304813]; Science Foundation of the Henan Province of China [20B310016]; Key R & D and Promotion Project in Henan Province [202102310101]; and Zhengzhou General Science and Technology Public Relations Project [141PPTGG449].