DLGAP5 通过激活 cAMP 促进胆囊癌迁移和肿瘤相关巨噬细胞 M2 极化。
DLGAP5 promotes gallbladder cancer migration and tumor-associated macrophage M2 polarization by activating cAMP.
机构信息
Department of Hepatobiliary Surgery, The Second Affiliated Hospital of Kunming Medical University, No. 374 Dianmian Avenue, Kunming, 650102, Yunnan, People's Republic of China.
出版信息
Cancer Immunol Immunother. 2023 Oct;72(10):3203-3216. doi: 10.1007/s00262-023-03484-6. Epub 2023 Jul 8.
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.
背景
尽管 disc large associated protein family（DLGAP5）已被报道参与多种肿瘤病理过程，但它在胆囊癌（GBC）中的表达和机制仍不确定。巨噬细胞分为 M1 和 M2 巨噬细胞。TAM 更被定义为 M2 极化的巨噬细胞，在癌症进展中发挥关键作用。
目的
阐明 disc large associated protein family（DLGAP5）在胆囊癌（GBC）进展中的作用，并探讨其机制。
方法
通过 R 语言分析 NCBI-GEO 中 GSE139682 数据库中 10 例正常癌旁组织和 10 例 GBC 组织中的差异基因。进行生物信息学分析和临床样本分析，检测 GBC 中 DLGAP5 的表达及其与预后的相关性。通过 CCK-8、EDU、Transwell、划痕愈合和免疫印迹检测 DLGAP5 对 GBC 细胞功能的影响。GST-pulldown 显示 DLGAP5 与 cAMP 的直接相互作用。进一步进行巨噬细胞极化实验，检测 DLGAP5 对巨噬细胞 M2 极化的影响。进一步进行肿瘤生长实验，以确认其在小鼠体内的作用。
结果
生物学分析和临床样本证实，DLGAP5 在 GBC 中升高，并与 GBC 患者的不良预后密切相关。在 GBC 细胞系如 GBC-SD 和 NOZ 细胞中转染 DLGAP5 后，细胞增殖和迁移增强，巨噬细胞向 M2 极化。然而，敲低 DLGAP5 后则出现相反的效果。机制上，DLGAP5 通过激活环磷酸腺苷（cAMP）通路促进 GBC-SD 和 NOZ 细胞的生长和迁移以及 THP-1 衍生巨噬细胞的 M2 极化。在体内，将 DLGAP5 敲低的 GBC-SD 细胞皮下注射到裸鼠体内。结果发现，敲低 DLGAP5 后，肿瘤体积和肿瘤均减小，与增殖和 M2 极化相关的指标降低。
结论
本研究表明，DLGAP5 在 GBC 中显著升高，与 GBC 患者的不良预后密切相关。DLGAP5 通过 cAMP 通路促进 GBC 增殖、迁移和巨噬细胞 M2 极化，为 GBC 的治疗提供了理论依据，可能成为有前途的治疗靶点。
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