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E2F5 Targeted by Let-7d-5p Facilitates Cell Proliferation, Metastasis and Immune Escape in Gallbladder Cancer

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Abstract

Background

Gallbladder cancer (GBC) remains a serious cause of cancer-related mortality across the globe. E2F5 has been identified to as a known oncogene in various cancers. However, the special functions of E2F5 have not been investigated in GBC.

Aims

To explore the regulatory functions of E2F5 and its related molecular regulatory mechanism in GBC progression.

Methods

The expression of genes were examined through qRT-PCR, western blot and IHC assay. The cell proliferation was assessed through CCK-8 and EDU assays. The cytotoxicity was tested through LDH assay. The percentage of CD8+ T cells and cell apoptosis were evaluated through flow cytometry. The binding ability was detected through luciferase reporter assay. The tumor growth was assessed through in vivo assays.

Results

In this study, it was demonstrated that E2F5 expression was evaluated in GBC, and resulted into poor prognosis. Bioinformatics analysis revealed E2F5 as a target for let-7d-5p, which when overexpressed, suppressed the metastasis and proliferation of GBC through the downregulation of E2F5. It was discovered that E2F5 activates JAK2/STAT3 signaling which is suppressed by let-7d-5p, implicating this pathway as one of the effectors of the oncogenic effects of ESF5 in GBC. E2F5 had been confirmed to aggravate tumor growth in vivo.

Conclusion

E2F5 targeted by let-7d-5p facilitated cell proliferation, metastasis and immune escape in GBC through the JAK2/STAT3 pathway.

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Funding

This work was supported by the China State Railway Group Co., Ltd. (J2022Z615).

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Author notes

  1. Lei Chen and Songyi Guo have contributed equally to this study.

Authors and Affiliations

  1. Department of Hepatobiliary Surgery, Peking University People’s Hospital, Beijing, 100044, China

    Lei Chen, Songyi Guo, Dafang Zhang & Xinyu Li

  2. Department of Hepatobiliary Oncology Surgery, Beijing Shijitan Hospital, Capital Medical University, No. 10, Tieyi Road, Yangfangdian, Haidian District, Beijing, 100038, China

    Jianfei Chen

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Contributions

LC, SYG wrote the manuscript, DFZ, XYL, JFC collected and analyzed the data. All authors read and approved the final manuscript.

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Correspondence to Jianfei Chen.

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Supplementary Information

Fig.S1 Knockdown or overexpression of E2F5 has no effects on the other members of the E2F family. A

The protein expressions of E2F1, E2F2, E2F6, E2F8 were examined after silencing E2F5 through WB. The other isoforms of E2F were not affected through downregulation via siRNA. B The protein expressions of E2F1, E2F2, E2F6, E2F8 were tested after overexpressing E2F5 through WB. The other isoforms of E2F were not affected after E2F5 overexpression. All the data are from three independent experiments (JPG 1854 kb)

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Chen, L., Guo, S., Zhang, D. et al. E2F5 Targeted by Let-7d-5p Facilitates Cell Proliferation, Metastasis and Immune Escape in Gallbladder Cancer. Dig Dis Sci 69, 463–475 (2024). https://doi.org/10.1007/s10620-023-08209-4

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