PO.TB10.03 · 肿瘤生物学
M2巨噬细胞在上皮性卵巢癌肿瘤微环境化疗耐药中的作用
Role of M2 macrophages in chemoresistance about the tumor microenvironment of epithelial ovarian cancer
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
目的：卵巢癌的标志性特征之一是对化疗药物耐药性的产生。与肿瘤细胞和基质微环境的相互作用促成了肿瘤相关巨噬细胞的表型极化。巨噬细胞至少由两个亚群组成，即M1和M2。M2巨噬细胞具备一系列促肿瘤能力，涉及免疫抑制、血管生成和新生血管形成，以及基质活化和重塑。因此，需要更实用的靶点来抑制由卵巢癌-巨噬细胞相互作用所致的化疗药物耐药和癌症进展。为了更好地理解卵巢癌细胞化疗耐药的机制，我们旨在研究巨噬细胞对肿瘤细胞卡铂反应的影响，并鉴定与化疗耐药相关的基因。
方法：通过对单核细胞进行细胞因子处理，将巨噬细胞分化为M1和M2巨噬细胞。通过transwell共培养实验修饰肿瘤微环境（TME）。我们施用卡铂并检测化疗耐药。我们还检测了与EMT（上皮-间质转化）、PD-L1和化疗耐药相关的关键基因的表达。Nanostring分析鉴定了与M2巨噬细胞共培养的卵巢癌细胞相较于单培养的肿瘤信号通路中的基因表达变化。进行了功能研究（增殖、迁移、侵袭、划痕愈合）以确定M2巨噬细胞在卵巢癌中的作用。
结果：与单培养的卵巢癌细胞相比，在与M2巨噬细胞共培养时iNOS下调，而PD-L1、CD206、TGF-b、MDR1、CSF-1和Arg1上调。PD-L1在M2巨噬细胞中也上调。与M2巨噬细胞共培养的卵巢癌细胞相较于单培养的卵巢癌细胞表现出更高的卡铂耐药性。促EMT相关基因CD2、VIM、ZEB1和SNAIL1在与M2巨噬细胞共培养的卵巢癌细胞中上调。Nanostring分析揭示了与肿瘤信号通路活化和EMT相关基因表达水平的变化。与M2巨噬细胞共培养的卵巢癌细胞进行功能研究，我们发现癌细胞的各项功能能力均显著增强。
结论：我们鉴定了与M2巨噬细胞共培养的卵巢癌细胞中多种基因表达的变化，并将PD-L1的上调确定为化疗耐药的关键因素。我们还进行了额外的Nanostring分析以细化M2巨噬细胞与卵巢癌细胞相互作用对肿瘤信号通路以及肿瘤微环境中化疗耐药的影响，并将分析这些数据并开展进一步实验。
查看英文原文 English abstract
OBJECTIVE One of the hallmark characteristics of ovarian cancer is the development of resistance to chemotherapeutics. The mutual interactions with tumor cells and stromal microenvironment contribute to phenotypically polarization of tumor associated macrophages. Macrophages consist of at least two subgroups, M1 and M2. M2 macrophages are endowed with a repertoire of tumor-promoting capabilities involving immuno-suppression, angiogenesis and neovascularization, as well as stromal activation and remodeling. Therefore, there is a need for more practical targets to inhibit chemotherapeutic drug resistance and cancer progression due to ovarian cancer-macrophage interactions. To better understand the mechanism of chemoresistance in ovarian cancer cells, we aimed to investigate the influence of macrophages on the tumor cell response to carboplatin and identify the genes associated with chemoresistance.
METHODS Macrophages were differentiated into M1 and M2 macrophages by cytokine treatment of monocytes. The tumor microenvironment (TME) was modified by transwell co-culture assay. We treated the carboplatin and tested chemoresistance. We also examined expression of key genes associated with EMT (Epithelial-Mesenchymal Transition), PD-L1 and chemoresistance. Nanostring analysis identified gene expression changes in tumor signaling pathways in ovarian cancer cells co-cultured with M2 macrophages compared to single-cultures. Functional study (proliferation, migration, invasion, wound healing) were performed to determine the role of the M2 macrophage in ovarian cancer.
RESULTS Compared to single-cultured ovarian cancer cells, iNOS was downregulated in co-cultured with M2 macrophages, while PD-L1,CD206,TGF-b,MDR1,CSF-1 and Arg1 were upregulated. PD-L1 was also upregulated in M2 macrophages. Ovarian cancer cells co-cultured with M2 macrophages showed higher carboplatin resistance compared to single-cultured ovarian cancer cells. EMT-promoting related genes CD2,VIM, ZEB1 and SNAIL1 were upregulated in ovarian cancer cells co-cultured with M2 macrophages. Nanostring analysis revealed changes in the expression levels of genes associated with tumor signaling pathway activation and EMT. Ovarian cancer cells co-cultured with M2 macrophages performed functional study, we found that the functional abilities of the cancer cells were all significantly increased.
CONCLUSION We identified changes in the expression of various genes in ovarian cancer cells which were co-cultured with M2 macrophages, and identified upregulation of PD-L1 as a key factor in chemoresistance. We have also conducted additional nanostrings to refine the impact of M2 macrophage and ovarian cancer cell interactions on tumour signalling pathways as well as chemoresistance in the tumour microenvironment, and will be analysing these data and conducting further experiments.
利益披露 Disclosure
S. Jeon, None.