Co-culturing polarized M2 Thp-1-derived macrophages enhance stemness of lung adenocarcinoma A549 cells.
Co-culturing polarized M2 Thp-1-derived macrophages enhance stemness of lung adenocarcinoma A549 cells.
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共培养极化的 M2 Thp-1 巨噬细胞增强肺腺癌 A549 细胞的干细胞性。
DOI：
10.21037/atm-21-1256
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发表时间：
2021-04
影响因子：
--
通讯作者：
Chen C
中科院分区：
文献类型：
--
作者：
Zhang X;Zhu M;Hong Z;Chen C
肿瘤微环境（TME）与癌症干细胞高度相关，并影响肿瘤的发生、进展和转移。本研究旨在探讨THP-1衍生的巨噬细胞诱导A549癌细胞干性的潜在分子机制。将Hedgehog抑制剂（Vismodegib）、Notch抑制剂γ分泌酶抑制剂（GSI）和信号转导器和转录激活剂3（STAT3）抑制剂葫芦素I（JSI-124）分别添加到A549癌细胞与THP-1源性巨噬细胞的共培养系统中。使用细胞计数试剂盒8（CCK-8）测定和Cell-IQ连续监测系统检查A549细胞的细胞生长和形态变化。分别通过定量实时聚合酶链式反应（qRT-PCR）和蛋白质印迹分析干细胞标志物的信使核糖核酸（mRNA）和蛋白质表达水平，并通过流式细胞术分析评估乙醛脱氢酶（ALDH）的活性。进行酶联免疫吸附测定（ELISA）和qRT-PCR测定来评估巨噬细胞的活化和分化。结果显示，与THP-1来源的巨噬细胞共培养后，A549细胞的增殖和干细胞性显着增强。与A549细胞共培养后，巨噬细胞中转化生长因子-β（TGF-β）和白细胞介素6（IL-6）的表达水平显着增加。同时，与 A549 细胞共培养诱导巨噬细胞向 M2 表型极化。此外，抑制剂可以减少共培养系统的增殖和干性，并降低TGF-β和IL-6的表达。这些结果表明，将 A549 细胞与 THP-1 衍生的巨噬细胞共培养可以通过多种促肿瘤途径诱导 A549 细胞的干性。因此，抑制巨噬细胞与肺癌干细胞之间的相互作用可能是未来肺癌治疗的可行目标。
The tumor microenvironment (TME) is highly associated with cancer stem cells, and affects tumor initiation, progression, and metastasis. This study aimed to explore the underlying molecular mechanism of induction of A549 cancer cell stemness by THP-1-derived macrophages. The Hedgehog inhibitor (Vismodegib), Notch inhibitor Gamma Secretase Inhibitor (GSI), and Signal Transducer and Activator of Transcription 3 (STAT3) inhibitor Cucurbitacin I (JSI-124) were added separately into the co-culture system of A549 cancer cell with THP-1-derived macrophages. Cell Counting Kit-8 (CCK-8) assay and the Cell-IQ continuous surveillance system were used to examine the cell growth and morphological changes of A549 cells. The messenger ribonucleic acid (mRNA) and protein expression levels of stem cell markers were respectively analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting, and the activity of Acetaldehyde dehydrogenase (ALDH) enzyme was assessed by flow cytometry analysis. Enzyme-linked immunosorbent assay (ELISA) and qRT-PCR assays were performed to evaluate the activation and differentiation of macrophages. Results showed that the proliferation and stemness of A549 cells were significantly enhanced by co-culturing with THP-1-derived macrophages. The expression levels of Transforming growth factor-β (TGF-β) and Interleukin-6 (IL-6) in macrophages were notably increased after co-culturing with A549 cells. Meanwhile, co-culturing with A549 cells induced the polarization of macrophages towards the M2 phenotype. Moreover, the inhibitors could reduce the proliferation and stemness of the co-culture system, and decrease the expression of TGF-β and IL-6. These results suggested that co-culturing A549 cells with THP-1-derived macrophages could induce the stemness of A549 cells via multiple pro-tumorigenic pathways. Thus, inhibition of the interaction between macrophages and lung cancer stem cells may be a viable target for lung cancer treatment in the future.
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