Type conversion of secretomes in a 3D TAM2 and HCC cell co-culture system and functional importance of CXCL2 in HCC.
Type conversion of secretomes in a 3D TAM2 and HCC cell co-culture system and functional importance of CXCL2 in HCC.
复制标题
3D TAM2 和 HCC 细胞共培养系统中分泌组的类型转换以及 CXCL2 在 HCC 中的功能重要性
DOI：
10.1038/srep24558
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发表时间：
2016-04-27
影响因子：
4.6
通讯作者：
Liu Y
中科院分区：
文献类型：
--
作者：
Lu Y;Li S;Ma L;Li Y;Zhang X;Peng Q;Mo C;Huang L;Qin X;Liu Y
巨噬细胞在肿瘤微环境中发挥重要作用，推动癌症进展和转移，特别是在肝细胞癌（HCC）中。然而，很少有研究评估了HCC中确切的分泌组组成。本研究探讨了不同表型的巨噬细胞对肝癌细胞的影响。M2能显著增强SMMC 7721细胞的增殖、迁移和侵袭能力（P均< 0.05）。然后将M2与SMMC 7721细胞共培养以重建肿瘤微环境。通过iTRAQ标记结合质谱分析，使用定量蛋白质组学分析来自M2、SMMC 7721细胞及其共培养系统的3D单一培养物的条件培养基。分泌组分析显示，与单一培养系统相比，共培养系统中总共有159种差异分泌蛋白，其中63种上调（>1.3倍），96种下调（<0.7倍）。CXCL2在共培养系统和HCC组织中被证实具有较高的表达，并被选择用于进一步研究。功能效应数据显示，重组人CXCL2显著增强SMMC 7721细胞的迁移、侵袭能力，减弱粘附能力。而CXCL2中和和CXCR2阻断则显著抑制了CXCL2对SMMC 7721细胞的作用，提示CXCL2可能在肝癌转移中起关键作用。
Macrophages play important roles in the tumor microenvironment, driving cancer progression and metastasis, particularly in hepatocellular carcinoma (HCC). However, few studies have assessed the exact secretome composition in HCC. In the present study, the impact of different phenotype of macrophages on HCC cells was investigated. Alternatively activated macrophages (M2) were found to significantly increase the proliferation, migration and invasion abilities of SMMC7721 cells (allP< 0.05). M2 were then co-cultured with SMMC7721 cells to reconstruct the tumor microenvironment. Conditioned medium from 3D single cultures of M2, SMMC7721 cells and their co-culture system were analyzed using quantitative proteomics via iTRAQ labeling combined with mass spectrometric analysis. Secretome analysis revealed a total of 159 differential secreted proteins in the co-culture system compared to the single culture systems, with 63 being up-regulated (>1.3-fold) and 96 down-regulated (<0.7-fold). CXCL2 was confirmed to have higher expression in the co-culture system and HCC tissues and was selected for further investigation. Functional effects data suggested that recombinant human CXCL2 significantly enhanced the migration, invasion ability of SMMC7721 cells and weakened adhesion ability. While CXCL2 neutralization and CXCR2 blockage significantly inhibited the effects of CXCL2 on SMMC7721 cells, indicating that CXCL2 may play pivotal role in HCC metastasis.