Tumor-associated macrophages regulate gastric cancer cell invasion and metastasis through TGFβ2/NF-κB/Kindlin-2 axis
Tumor-associated macrophages regulate gastric cancer cell invasion and metastasis through TGFβ2/NF-κB/Kindlin-2 axis
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DOI：
10.21147/j.issn.1000-9604.2020.01.09
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发表时间：
2020-02-01
影响因子：
5.1
通讯作者：
Wang, Shan
中科院分区：
文献类型：
--
作者：
Wang, Zhu;Yang, Yang;Wang, Shan
目的：最近的研究表明肿瘤相关巨噬细胞（TAM）在癌症侵袭和转移中发挥重要作用。我们前期的研究报道TAMs通过Kindlin-2通路促进胃癌（GC）细胞的侵袭和转移。但其机制有待阐明。方法：采用PMA/白介素(IL)-4/IL-13诱导THP-1单核细胞建立高效的体外TAM模型，并通过流式细胞仪分离M2型巨噬细胞。使用双荧光素酶报告系统和染色质免疫沉淀 (ChIP) 测定来研究转化生长因子 β 2 (TGF β 2) 调节 Kindlin-2 表达的机制。采用免疫组织化学方法研究人胃癌组织中TAM浸润、Kindlin-2蛋白表达、临床病理参数与预后的关系。采用裸鼠肿瘤发生模型验证体内侵袭和转移机制。结果：我们发现与TAMs共培养的GC细胞中Kindlin-2的mRNA和蛋白水平表达上调，与较高的侵袭率相关。 Kindlin-2敲低降低了共培养条件下GC细胞的侵袭率。 TAM 分泌的 TGF beta 2 通过转录因子 NF-kappa B 调节 Kindlin-2 的表达。因此，TAM 通过 TGF beta 2/NF-kappa B/Kindlin-2 轴参与 GC 的进展。 Kindlin-2表达和TAM浸润与TNM分期显着正相关，Kindlin-2高表达患者的总生存率显着低于Kindlin-2低表达患者。此外，Kindlin-2在体内促进GC细胞的侵袭。结论：本研究阐明了TAMs通过TGF beta 2/NF-kappa B/Kindlin-2轴参与GC细胞侵袭和转移的机制，为新的治疗选择和方法提供了可能。
Objective: Recent studies have shown that tumor-associated macrophages (TAMs) play an important role in cancer invasion and metastasis. Our previous studies have reported that TAMs promote the invasion and metastasis of gastric cancer (GC) cells through the Kindlin-2 pathway. However, the mechanism needs to be clarified.Methods: THP-1 monocytes were induced by PMA/interleukin (IL)-4/IL-13 to establish an efficient TAM model in vitro and M2 macrophages were isolated via flow cytometry. A dual luciferase reporter system and chromatin immunoprecipitation (ChIP) assay were used to investigate the mechanism of transforming growth factor beta 2 (TGF beta 2) regulating Kindlin-2 expression. Immunohistochemistry was used to study the relationships among TAM infiltration in human GC tissues, Kindlin-2 protein expression, clinicopathological parameters and prognosis in human GC tissues. A nude mouse oncogenesis model was used to verify the invasion and metastasis mechanisms in vivo.Results: We found that Kindlin-2 expression was upregulated at both mRNA and protein levels in GC cells cocultured with TAMs, associated with higher invasion rate. Kindlin-2 knockdown reduced the invasion rate of GC cells under coculture condition. TGF beta 2 secreted by TAMs regulated the expression of Kindlin-2 through the transcription factor NF-kappa B. TAMs thus participated in the progression of GC through the TGF beta 2/NF-kappa B/Kindlin-2 axis. Kindlin-2 expression and TAM infiltration were significantly positively correlated with TNM stage, and patients with high Kindlin-2 expression had significantly poorer overall survival than patients with low Kindlin-2 expression. Furthermore, Kindlin-2 promoted the invasion of GC cells in vivo.Conclusions: This study elucidates the mechanism of TAMs participating in GC cell invasion and metastasis through the TGF beta 2/NF-kappa B/Kindlin-2 axis, providing a possibility for new treatment options and approaches.