逆转肿瘤相关巨噬细胞的极化可抑制肿瘤转移。

Reversing the polarization of tumor-associated macrophages inhibits tumor metastasis.

作者信息

Yuan Rufei, Li Shengfu, Geng Hua, Wang Xin, Guan Qinglin, Li Xiaohuan, Ren Changle, Yuan Xiaolin

机构信息

Center Laboratory, Affiliated Zhongshan Hospital of Dalian University, Dalian 116001, China.

Orthopedic Surgery Department, Dalian Municipal Central Hospital, Dalian 116033, China.

出版信息

Int Immunopharmacol. 2017 Aug;49:30-37. doi: 10.1016/j.intimp.2017.05.014. Epub 2017 May 25.

DOI:10.1016/j.intimp.2017.05.014

PMID:28550732

Abstract

OBJECTIVE

The M2 phenotype is dominant in tumor associated macrophages (TAM), and plays a key role in promoting tumor growth, invasion and metastasis. Converting TAM polarization from M2 to M1 may contribute to eliciting anti-tumor-specific immune responses and inhibiting tumor metastasis. In this study, the effect of reversing the polarization of TAM on tumor metastasis was investigated.

METHODS

Peritoneal macrophages were obtained from BABL/c mice, and M2 polarization was induced by IL-4. In an in vivo experiment, BABL/c mice were transplanted with 4T1 tumor cells. In vitro and in vivo experimental studies, M2 macrophage polarization was reversed with CpG-DNA or CpG-DNA combined with anti-IL-10R Ab. CD68, MHCII and FRβ molecular expression in macrophages were examined with immunofluorescence staining. The mRNA expression of IL-2, IL-6, IL-13, VEGF and MMP-9 were detected with RT-PCR. VEGF and MMP-9 protein expression of tumors in situ was measured by western blot assay. Lung-metastasis of the tumor was observed and assessed by micro-CT.

RESULTS

CpG-DNA and CpG-DNA combined with anti-IL-10R Ab could promote MHCII, IL-2, IL-6 and IL-13 molecular expression, and suppress the expression of FRβ, MMP-9 and VEGF, in both freshly isolated peritoneal macrophages and M2 macrophages. In the CpG-DNA combined with anti-IL-10R Ab injecting group, the percentage of CD68 MHCII cells were significantly higher than that of CD68FRβ cells (P<0.05). This was distinct from the result of the control group, which CD68 FRβ was higher than CD68MHCIIcells (P<0.01). Furthermore, VEGF-A and MMP-9 level in primary tumor tissues in the experimental group was significantly lower (P<0.01), compared to the control group. Moreover, the number of detectable lung-metastasis foci was significantly lower in the experimental group than in the control group (P<0.05).

CONCLUSION

Reversing the polarization of TAM from M2 to M1 phenotype can inhibit tumor metastasis.

摘要

目的

M2表型在肿瘤相关巨噬细胞（TAM）中占主导地位，在促进肿瘤生长、侵袭和转移中起关键作用。将TAM极化从M2转变为M1可能有助于引发抗肿瘤特异性免疫反应并抑制肿瘤转移。本研究探讨了逆转TAM极化对肿瘤转移的影响。

方法

从BABL/c小鼠获取腹腔巨噬细胞，用IL-4诱导M2极化。在体内实验中，将4T1肿瘤细胞移植到BABL/c小鼠体内。在体外和体内实验研究中，用CpG-DNA或CpG-DNA联合抗IL-10R抗体逆转M2巨噬细胞极化。用免疫荧光染色检测巨噬细胞中CD68、MHCII和FRβ分子表达。用RT-PCR检测IL-2、IL-6、IL-13、VEGF和MMP-9的mRNA表达。用蛋白质印迹法检测肿瘤原位VEGF和MMP-9蛋白表达。通过微型CT观察和评估肿瘤的肺转移情况。

结果

CpG-DNA以及CpG-DNA联合抗IL-10R抗体可促进新鲜分离的腹腔巨噬细胞和M2巨噬细胞中MHCII、IL-2、IL-6和IL-13分子表达，并抑制FRβ、MMP-9和VEGF的表达。在CpG-DNA联合抗IL-10R抗体注射组中，CD68 MHCII细胞百分比显著高于CD68FRβ细胞（P<0.05）。这与对照组结果不同，对照组中CD68 FRβ高于CD68MHCII细胞（P<0.01）。此外，与对照组相比，实验组原发肿瘤组织中VEGF-A和MMP-9水平显著降低（P<0.01）。而且，实验组可检测到的肺转移灶数量明显低于对照组（P<0.05）。