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Cellular and Molecular Biology

The CBS-H2S axis promotes liver metastasis of colon cancer by upregulating VEGF through AP-1 activation

British Journal of Cancer volume 126, pages 1055–1066 (2022)Cite this article

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Abstract

Background

The main therapy for colon cancer with liver metastasis is chemotherapy based on 5-fluorouracil combined with targeted drugs. However, acquired drug resistance and severe adverse reactions limit patients’ benefit from standard chemotherapy. Here, we investigate the involvement of endogenous hydrogen sulfide (H2S) in liver metastasis of colon cancer and its potential value as a novel therapeutic target.

Methods

We used the CRISPR/Cas9 system to knockdown CBS gene expression in colon cancer cell lines. PCR arrays and proteome arrays were applied to detect the transcription and protein expression levels, respectively, of angiogenesis-related genes after knockdown. The molecular mechanism was investigated by western blot analysis, RT–qPCR, immunofluorescence staining, ChIP assays and dual-luciferase reporter assays. A liver metastasis mouse model was adopted to investigate the effect of targeting CBS on tumour metastasis in vivo.

Results

Knockdown of CBS decreased the metastasis and invasion of colon cancer cells and inhibited angiogenesis both in vivo and in vitro. Tissue microarray analysis showed a positive correlation between CBS and VEGF expression in colon cancer tissues. Further analysis at the molecular level validated a positive feedback loop between the CBS-H2S axis and VEGF.

Conclusions

Endogenous H2S promotes angiogenesis and metastasis in colon cancer, and targeting the positive feedback loop between the CBS-H2S axis and VEGF can effectively intervene in liver metastasis of colon cancer.

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Fig. 1: Effects of CBS inhibition on H2S production, invasion and metastasis in SW480 and DLD1 cells.
Fig. 2: Effect of CBS inhibition on angiogenesis of colon cancer cells.
Fig. 3: The correlation between CBS and VEGF expression in colon cancer tissues.
Fig. 4: The effect of the CBS-H2S axis on the activation of VEGF and the effect of VEGF on the activation of CBS.
Fig. 5: The effect of CBS and VEGF on liver metastasis of colon cancer cells in vivo.

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Data availability

The data underlying this article are available in the article and in its online supplementary material.

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Acknowledgements

We would like to thank Professor Yi Long and Professor Ding-fang Bu for their excellent technical assistance.

Funding

This research was supported by the National Natural Science Foundation of China (No. 81902384).

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Authors and Affiliations

  1. Division of General Surgery, Peking University First Hospital, Peking University, 100034, Beijing, People’s Republic of China

    Shihao Guo, Jichang Li, Zhihao Huang, Taohua Yue, Jing Zhu, Xin Wang, Yucun Liu, Pengyuan Wang & Shanwen Chen

  2. Department of Colorectal Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China

    Shihao Guo

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Contributions

SG: designed the study; performed experiments; wrote, reviewed and edited the manuscript. JL: performed experiments. ZH: performed experiments. TY: performed experiments. JL: performed experiments. JZ: provided resources. XW: provided resources. YL: designed the study. PW: designed the study; wrote, reviewed and edited the manuscript. SC: designed the study; performed experiments; wrote, reviewed and edited the manuscript.

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Correspondence to Shanwen Chen.

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Ethics approval statement

The experimental protocol complied with the Guide for Care and Use of Laboratory Animals and was authorised by the Institutional Animal Care and Use Committee of Peking University for the use of experimental animals (No. J201965). All animal studies were complied with relevant ethical regulations for animal testing and research.

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Guo, S., Li, J., Huang, Z. et al. The CBS-H2S axis promotes liver metastasis of colon cancer by upregulating VEGF through AP-1 activation. Br J Cancer 126, 1055–1066 (2022). https://doi.org/10.1038/s41416-021-01681-7

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