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DLGAP5 promotes gallbladder cancer migration and tumor-associated macrophage M2 polarization by activating cAMP

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Abstract

Background

Although disc large associated protein family (DLGAP5) has been reported to be involved in a variety of tumor pathologic processes, its expression and mechanism in gallbladder cancer (GBC) are still uncertain. Macrophages were divided into M1 and M2 macrophages. TAM is more closely defined as M2 polarized macrophages, which plays a key role in cancer progression.

Objective

To clarify the role of disc large associated protein family (DLGAP5) in gallbladder cancer (GBC) progression and investigate the mechanism.

Methods

Differential genes in 10 normal paracancer tissues and 10 GBC tissues in GSE139682 from NCBI-GEO were analyzed by R language. Bioinformation analysis and clinical sample analysis were performed to detect DLGAP5 expression in GBC and its correlation with prognosis. CCK-8, EDU, transwell, wound closure, and Immunoblot were performed to detect its effects on the function of GBC cells. GST-pulldown showed the direct interact between DLGAP5 and cAMP. Macrophage polarization assay was further conducted to detect the effects of DLGAP5 on macrophage M2 polarization. The tumor growth assays were further conducted to confirm its role in mice.

Results

Biological analysis and clinical samples confirmed that DLGAP5 was increased in GBC and strongly related to poor prognosis in patients with GBC. After overexpression of DLGAP5 in GBC cell lines, such as GBC-SD and NOZ cells, cell proliferation and migration were enhanced, and macrophages were polarized to M2. However, after DLGAP5 is knocked down, there is opposite effect. Mechanistically, DLGAP5 promotes the growth and migration of GBC-SD and NOZ cells and the M2 polarization of THP-1-derived macrophages by activating cyclic adenosine monophosphate (cAMP) pathway. In vivo, GBC-SD with DLGAP5 knockdown was subcutaneously injected into nude mice. It was found that after DLGAP5 knockdown, both tumor volume and tumor were reduced, and indicators related to proliferation and M2 polarization decreased.

Conclusion

Our study shows that DLGAP5 is significantly elevated in GBC and is strongly related to poor prognosis in patients with GBC. DLGAP5 promotes GBC proliferation, migration, and M2 polarization of macrophages through cAMP pathway, which provides a theoretical basis for the treatment of GBC and may become a promising therapeutic target.

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Data availability

All data generated or analyzed during this study are included in this published article.

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Funding

This work was supported by the Yun nan Fundamental Research Projects (Grant No.202101AT070239) and Medical Reserve Talents project of Yunnan Provincial Health Commsion (Grant No. H-2018065).

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Authors and Affiliations

  1. Department of Hepatobiliary Surgery, The Second Affiliated Hospital of Kunming Medical University, No. 374 Dianmian Avenue, Kunming, 650102, Yunnan, People’s Republic of China

    Jie Huang, Mengyao Zheng, Yan Li, Dingwei Xu & Daguang Tian

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  1. Jie Huang
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  2. Mengyao Zheng
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  3. Yan Li
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  4. Dingwei Xu
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Contributions

All authors contributed to the study conception and design. Material preparation and the experiments were performed by Jie Huang. Data collection and analysis were performed by Mengyao Zheng, and Yan Li. The first draft of the manuscript was written by Dingwei Xu and Daguang Tian and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Jie Huang.

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All procedures performed in studies involving human participants were in accordance with the standards upheld by the Ethics Committee of the Second Affiliated Ho spital of Kunming Medical University and with those of the 1964 Helsinki Declaration and its later amendments for ethical research involving human subjects. All animal experiments were approved by the Ethics Committee of the Second Affiliated Hospital of Kunming Medical University for the use of animals and conducted in accordance with the National Institutes of Health Laboratory Animal Care and Use Guidelines.

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Huang, J., Zheng, M., Li, Y. et al. DLGAP5 promotes gallbladder cancer migration and tumor-associated macrophage M2 polarization by activating cAMP. Cancer Immunol Immunother 72, 3203–3216 (2023). https://doi.org/10.1007/s00262-023-03484-6

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