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Establishment of SLC7A11‐knockout mouse and its preliminary investigation in melanoma

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Abstract

Solute carrier family 7 member 11 (SLC7A11)/xCT is an amino acid transporter that mediates the cystine uptake and glutamate export, participates in several malignant tumors’ progression. However, the role of SLC7A11 on the occurrence and development of melanoma still remains unclear. Here, the transcribed mRNA encoding for Cas9 and sgRNA targeting SLC7A11 in vitro were microinjected into zygotes, to establish the SLC7A11 knockout (KO) mice (SLC7A11−/−). Further, we conducted melanoma-bearing mice using the metastatic melanoma cell line (B16-F10) to observe the melanoma development. There was no off-target in KO mice detected by T7E1 cleavage assay. The results showed that the tumor volume of KO mice was significantly lower than that of SLC7A11+/+ (WT) mice at 8d, 10d, 12d, 14d, and 16d (P < 0.05). The tumors of WT appeared to more disorganized morphology, more unbalanced nuclear-cytoplasmic ratio, less defined boundary, and increased tumor necrosis. And after SLC7A11 deletion, the expression of CXCL9 and TLR6 were significantly up-regulated, and that of NOS2 and CCL8 were significantly down-regulated (P < 0.01). Additionally, Ki67 immunostaining revealed lower proliferating cells in the tumors of SLC7A11 KO mice compared to WT mice. In summary, the deletion of SLC7A11 significantly inhibited the development of melanoma. Our results provide direct evidence to identify SLC7A11 as a novel target for molecular therapy and prognosis judgment of melanoma.

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Data availability

The data supporting the findings of this study are available in https://doi.org/10.6084/m9.figshare.23513307.

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Acknowledgements

The research was financially supported by the National Natural Science Foundation of China (Grant no. 32172722), and Yangzhou University Student Innovation and Entrepreneurship Training Program Project (Grant no. X20220652) and Qing Lan Project of Yangzhou university.

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Authors and Affiliations

  1. College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, China

    Yang Chen, Tingting Lu, Yufei Liu, Yongqi Liu, Shaocheng Bai, Qiuran Chen, Bohao Zhao & Xinsheng Wu

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  1. Yang Chen
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  2. Tingting Lu
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  3. Yufei Liu
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  8. Xinsheng Wu
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Contributions

Yang Chen: Data curation, Formal analysis, Methodology, Writing—review & editing. Tingting Lu, Yufei Liu, Yongqi Liu: Data curation, Formal analysis, Methodology. Shaocheng Bai, Qiuran Chen, Bohao Zhao: Formal analysis, Software, Validation. Xinsheng Wu: Data curation, Funding acquisition, Supervision. All authors contributed to, edited, and approved the final manuscript as submitted.

Corresponding author

Correspondence to Xinsheng Wu.

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Ethics approval

The animal experiments were approved by the Animal Ethics Committee of Yangzhou University, China (approval No. 201802126) and were strictly implemented according to the Guide for the Care and Use of Laboratory Animals. The mice were sacrificed by cervical dislocation following anesthesia with Shutai.

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The authors declare no conflict of interest.

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Chen, Y., Lu, T., Liu, Y. et al. Establishment of SLC7A11‐knockout mouse and its preliminary investigation in melanoma. In Vitro Cell.Dev.Biol.-Animal 59, 729–737 (2023). https://doi.org/10.1007/s11626-023-00819-6

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  • DOI: https://doi.org/10.1007/s11626-023-00819-6

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