CCL2/CCR2 Axis Promotes the Progression of Salivary Adenoid Cystic Carcinoma via Recruiting and Reprogramming the Tumor-Associated Macrophages
CCL2/CCR2 Axis Promotes the Progression of Salivary Adenoid Cystic Carcinoma via Recruiting and Reprogramming the Tumor-Associated Macrophages
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DOI：
10.3389/fonc.2019.00231
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发表时间：
2019-04-09
影响因子：
4.7
通讯作者：
Yang, Xinjie
中科院分区：
文献类型：
--
作者：
Yang, Zihui;Li, Huan;Yang, Xinjie
目的：探讨CCL2/CCR2轴在涎腺腺样囊性癌(SACC)进展过程中肿瘤细胞与肿瘤相关巨噬细胞(TAM)相互作用中的作用及其机制。方法：采用免疫组织化学染色和生存分析方法，研究CD68、CD163、CCL2、CCR2在涎腺腺样囊性癌(SACC)中的表达及其临床意义。用RNA干扰技术沉默CCL2，用CCR2特异性拮抗剂(RS504393)阻断CCR2。采用酶联免疫吸附试验、定量逆转录聚合酶链式反应、免疫印迹、免疫荧光、流式细胞仪、CCK8、划痕愈合、Transwell等方法，探讨CCL2/CCR2轴在SACC细胞与TAMs相互作用中的作用及可能机制。结果：CD68标记的TAMS和CD163标记的M2 TAMS在SACC组织中的高表达与CCL2和CCR2的表达显著相关。值得注意的是，TAMS的高浸润性和CCL2的过度表达与SACC的临床进展和预后不良明显相关。SACC细胞来源的CCL2在体外可激活其受体CCR2在TAMs中的表达。体外实验结果进一步表明，CCL2来源的SACC细胞通过CCL2/CCR2轴参与TAMs的募集、M2极化和GDNF的表达。同时，TAMS衍生的GDNF通过GDNF/p-RET途径促进SACC细胞的增殖、迁移和侵袭。CCR2拮抗剂(RS504393)治疗免疫缺陷小鼠可显著抑制TAMs的侵袭和SACC细胞的致瘤性。结论：CCL2/CCR2轴通过对TAMs的募集和重新编程促进SACC细胞的进展。通过阻断CCL2/CCR2轴靶向TAMs可能是SACC治疗的一种有前景的策略。
Objective: The present study investigated the roles and underlying mechanism of CCL2/CCR2 axis in the interactions between tumor cells and tumor-associated macrophages (TAMs) during the progression of salivary adenoid cystic carcinoma (SACC).Methods: Immunohistochemical staining and survival analysis were performed to study the correlation and clinical value of CD68, CD163, CCL2, and CCR2 expression in SACC cases. CCL2 silencing by RNA interference and CCR2 blocking by CCR2 specific antagonist (RS504393) were performed. ELISA, qRT-PCR, western blot, immuno fluorescence, flow cytometry, CCK8, scratch wound healing, and transwell assays were used to explore the functional roles and possible mechanism of CCL2/CCR2 axis in the interactions between SACC cells and TAMs. The effects of targeting TAMs by blocking the CCL2/CCR2 axis were investigated in a xenograft mice model with SACC cells.Results: The high infiltration of TAMs marked by CD68 and high infiltration of M2 TAMs marked by CD163 were significantly correlated with the expression of CCL2 and CCR2 in SACC tissues. Notably, the high infiltration of TAMs and the overexpression of CCL2 were obviously associated with the clinical progression and poor prognosis of SACC. SACC cells derived CCL2 could activate its receptor CCR2 expression in TAMs in vitro. The in vitro results further indicated that the SACC cells derived CCL2 was involved in the recruitment, M2 polarization, and GDNF expression of TAMs through the CCL2/CCR2 axis. Meanwhile, TAMs derived GDNF promoted the proliferation, migration, and invasion of SACC cells through the GDNF/p-RET pathway. Treating immunodeficient mice with the CCR2 antagonist (RS504393) greatly inhibited the infiltration of TAMs and the tumorigenicity of SACC cells.Conclusion: These new findings indicated that the CCL2/CCR2 axis promoted the progression of SACC cells via recruiting and reprogramming TAMs. Targeting TAMs by blocking the CCL2/CCR2 axis might be a prospective strategy for SACC therapy.