将极化的M2型Thp-1衍生巨噬细胞进行共培养可增强肺腺癌A549细胞的干性。

Co-culturing polarized M2 Thp-1-derived macrophages enhance stemness of lung adenocarcinoma A549 cells.

作者信息

Zhang Xiaocheng, Zhu Mingyang, Hong Zipu, Chen Chengshui

机构信息

Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China.

Department of Traumatology, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, China.

出版信息

Ann Transl Med. 2021 Apr;9(8):709. doi: 10.21037/atm-21-1256.

BACKGROUND

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.

METHOD

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

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.

CONCLUSIONS

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.

背景

肿瘤微环境（TME）与癌症干细胞高度相关，并影响肿瘤的起始、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。

方法

将刺猬信号通路抑制剂（维莫德吉）、Notch抑制剂γ-分泌酶抑制剂（GSI）和信号转导子及转录激活子3（STAT3）抑制剂葫芦素I（JSI-124）分别加入A549癌细胞与THP-1衍生的巨噬细胞的共培养体系中。使用细胞计数试剂盒-8（CCK-8）检测法和Cell-IQ连续监测系统检测A549细胞的生长和形态变化。通过定量实时聚合酶链反应（qRT-PCR）和蛋白质印迹法分别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检测以评估巨噬细胞的激活和分化。

结果

结果显示，与THP-1衍生的巨噬细胞共培养可显著增强A549细胞的增殖和干性。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）的表达水平显著升高。同时，与A549细胞共培养诱导巨噬细胞向M2表型极化。此外，这些抑制剂可降低共培养体系的增殖和干性，并降低TGF-β和IL-6的表达。