College of Pharmacy, Zhejiang Chinese Medical University.
Department of Thoracic Surgery, Zhejiang Hospital, Hangzhou, China.
Anticancer Drugs. 2024 Jan 1;35(1):22-35. doi: 10.1097/CAD.0000000000001538. Epub 2023 Aug 25.
This study aimed to explore the interaction between the tumor-associated macrophage (TAM) and enhancer of zeste homolog 2 (EZH2) in tumor microenvironment of lung cancer are obscure. M2 type of TAM was induced by interleukin-4 (IL-4) and interleukin-13 (IL-13) in RAW264.7 cells. Subsequently, the co-culture system of the M2 RAW264.7 treating LLC-1 cells were constructed to evaluate the cell proliferation, migration and invasion abilities. On top of that, the M2 RAW264.7 was injected into the LLC-1 cells-bearing mice. Tumor growth and the number of metastatic nodes were observed. Moreover, DNA methylation, EZH2 expression, target genes of EZH2 and the M2 type TAM-related markers were detected in vivo and in vitro . Further experiments of EZH2 function in lung cancer were carried out by the addition of EZH2 inhibitor (GSK126) and si-EZH2. M2 type of TAM was induced with IL-4 and IL-13 with increased expression of CD206, CD68, CD163 and Arg1. Following co-culture with M2 type TAM, the proliferative, invasive, migrative abilities, tumor growth and metastasis, and the DNA methylation, EZH2 level were strengthened whereas the target genes of EZH2, including p21, CDKN2A, CDKN2B were reduced in LLC-1 cells and LLC-1 cell-bearing mice. Of note, GSK126 and si-EZH2 offset the M2 type TAM's effects, and inhibited the LLC-1 cell metastasis, DNA methylation and tumor growth. M2 type TAM promoted DNA methylation in LLC-1 cells and LLC-1 cell-bearing mice, which is related to the intensified EZH2.
这项研究旨在探讨肿瘤相关巨噬细胞（TAM）与增强子的锌指蛋白 2（EZH2）在肺癌肿瘤微环境中的相互作用尚不清楚。用白细胞介素 4（IL-4）和白细胞介素 13（IL-13）诱导 RAW264.7 细胞产生 M2 型 TAM。随后，构建 M2 RAW264.7 处理 LLC-1 细胞的共培养体系，评估细胞增殖、迁移和侵袭能力。此外，将 M2 RAW264.7 注射到携带 LLC-1 细胞的小鼠体内，观察肿瘤生长和转移节点数量。此外，还在体内和体外检测了 DNA 甲基化、EZH2 表达、EZH2 的靶基因和 M2 型 TAM 相关标志物。通过添加 EZH2 抑制剂（GSK126）和 si-EZH2 进一步进行肺癌中 EZH2 功能的实验。用白细胞介素 4 和白细胞介素 13 诱导 M2 型 TAM，其 CD206、CD68、CD163 和 Arg1 表达增加。与 M2 型 TAM 共培养后，LLC-1 细胞的增殖、侵袭、迁移能力、肿瘤生长和转移，以及 EZH2 水平增强，而 EZH2 的靶基因，包括 p21、CDKN2A 和 CDKN2B 降低。值得注意的是，GSK126 和 si-EZH2 抵消了 M2 型 TAM 的作用，并抑制了 LLC-1 细胞的转移、DNA 甲基化和肿瘤生长。M2 型 TAM 促进了 LLC-1 细胞和携带 LLC-1 细胞的小鼠的 DNA 甲基化，这与 EZH2 的增强有关。
