Department of Physiology, Chungnam National University College of Medicine, Daejeon, Republic of Korea.
Department of thoracic surgery, Chungnam National University School of Medicine, Daejeon, Republic of Korea.
BMC Mol Cell Biol. 2024 Mar 4;25(1):6. doi: 10.1186/s12860-024-00502-y.
Macrophages promote angiogenesis, metastasis, and drug resistance in several cancers. Similarly, TonEBP/NFAT5 induces metastasis in renal carcinoma and colon cancer cells. However, the role of this transcription factor and that of macrophages in lung cancer cells remains unclear. Therefore, this study investigated the effects of macrophages and TonEBP/NFAT5 expression on cisplatin resistance and migration in A549 lung adenocarcinoma cells.
A549 cells were cultured alone or indirectly co-cultured with THP-1-derived macrophages using a transwell culture chamber. Cisplatin-induced cell death was markedly decreased and migration increased in co-cultured A549 cells. Macrophage-conditioned media (CM) showed a similar effect on drug resistance and migration. Cisplatin-induced apoptosis, DNA fragmentation, and cleaved apoptotic proteins PARP and caspase-3 were markedly reduced in macrophage CM-induced A549 cells. Here, ERK, p38, JNK, and NF-κB activities were increased by macrophage CM. Furthermore, the proteins involved in cisplatin resistance and cancer cell migration were identified using specific inhibitors of each protein. ERK and NF-κB inhibition considerably reduced cisplatin resistance. The increase in macrophage CM-induced migration was partially reduced by treatment with ERK, JNK, and NF-κB inhibitors. TonEBP/NFAT5 expression was increased by macrophages, resulting in increased cisplatin resistance, cell migration, and invasion. Moreover, RNAi-mediated knockdown of TonEBP/NFAT5 reduced cisplatin resistance, migration, and invasion in macrophage CM-induced A549 cells.
These findings demonstrate that paracrine factors secreted from macrophages can change A549 cells, resulting in the induction of drug resistance against cisplatin and migration. In addition, the TonEBP/NFAT5 ratio, increased by macrophages, is an important regulator of the malignant transformation of cells.
巨噬细胞可促进多种癌症中的血管生成、转移和耐药性。同样，TonEBP/NFAT5 可诱导肾细胞癌和结肠癌细胞转移。然而，该转录因子和巨噬细胞在肺癌细胞中的作用尚不清楚。因此，本研究探讨了巨噬细胞和 TonEBP/NFAT5 表达对 A549 肺腺癌细胞顺铂耐药性和迁移的影响。
A549 细胞单独培养或通过 Transwell 培养室与 THP-1 衍生的巨噬细胞间接共培养。共培养的 A549 细胞中，顺铂诱导的细胞死亡明显减少，迁移增加。巨噬细胞条件培养基（CM）对耐药性和迁移也有类似的影响。巨噬细胞 CM 诱导的 A549 细胞中，顺铂诱导的细胞凋亡、DNA 片段化和裂解的凋亡蛋白 PARP 和 caspase-3 明显减少。在此，巨噬细胞 CM 增加了 ERK、p38、JNK 和 NF-κB 的活性。此外，使用每种蛋白的特异性抑制剂鉴定了与顺铂耐药性和癌细胞迁移相关的蛋白。ERK 和 NF-κB 抑制可显著降低顺铂耐药性。用 ERK、JNK 和 NF-κB 抑制剂处理可部分降低巨噬细胞 CM 诱导的迁移增加。巨噬细胞增加了 TonEBP/NFAT5 的表达，导致顺铂耐药性、细胞迁移和侵袭增加。此外，RNAi 介导的 TonEBP/NFAT5 敲低可降低巨噬细胞 CM 诱导的 A549 细胞中的顺铂耐药性、迁移和侵袭。
这些发现表明，巨噬细胞分泌的旁分泌因子可以改变 A549 细胞，从而诱导顺铂耐药性和迁移。此外，巨噬细胞增加的 TonEBP/NFAT5 比率是细胞恶性转化的重要调节剂。
