Abstract：
Melanoma is a prevalent malignant skin tumor known for its high invasive ability and a high rate of metastasis, making clinical treatment exceptionally challenging. Tumor-associated macrophages (TAMs) are the most abundant immune cells in the tumor microenvironment and play a crucial role in tumor survival and development. Cold atmospheric plasma (CAP) is an emerging tool for tumor treatment that has garnered attention from scholars due to its interaction with non-tumor cells in the tumor microenvironment. Here, we used the macrophage lines THP-1 and RAW264.7, as well as the melanoma cell lines A375 and MV3, as research subjects to investigate the effect of plasma-activated liquid (PAL) on macrophage differentiation and its inhibitory effect on melanoma cell proliferation. We confirmed that the killing effect of PAL on melanoma cells was selective. Using flow cytometry and PCR, we discovered that PAL can influence macrophage differentiation. Through in vitro cell coculture, we demonstrated that PAL-treated macrophages can significantly impede tumor cell development and progression, and the effect is more potent than that of PAL directly targeting tumor cells. Furthermore, we have proposed the hypothesis that PAL promotes the differentiation of macrophages into the M1 type through the ROS/JAK2/STAT1 pathway. To test the hypothesis, we employed catalase and fludarabine to block different sites of the pathway. The results were then validated through Western Blot, qPCR and ELISA. This study illustrates that PAL therapy is an effective tumor immunotherapy and expands the scope of tumor immunotherapy. Furthermore, these findings establish a theoretical foundation for potential clinical applications of PAL.
摘要：
黑色素瘤是一种常见的恶性皮肤肿瘤，以其高侵袭能力和高转移率而著称。使临床治疗异常具有挑战性。肿瘤相关巨噬细胞(TAMs)是肿瘤微环境中最丰富的免疫细胞,在肿瘤的生存和发展中起着至关重要的作用。冷大气等离子体（CAP）是一种新兴的肿瘤治疗工具，由于其与非肿瘤细胞在肿瘤微环境中的相互作用而引起了学者的关注。这里,我们以巨噬细胞系THP-1和RAW264.7,以及黑色素瘤细胞系A375和MV3为研究对象,研究血浆活化液体(PAL)对巨噬细胞分化的影响及其对黑色素瘤细胞增殖的抑制作用.我们证实PAL对黑色素瘤细胞的杀伤感化是选择性的。使用流式细胞术和PCR，我们发现PAL可以影响巨噬细胞的分化。通过体外细胞共培养，我们证明，PAL处理的巨噬细胞可以显著阻碍肿瘤细胞的发展和进展，该效应比直接靶向肿瘤细胞的PAL更有效。此外，我们提出了以下假设：PAL通过ROS/JAK2/STAT1途径促进巨噬细胞分化为M1型。为了检验假设，我们使用过氧化氢酶和氟达拉滨阻断该途径的不同位点。然后通过WesternBlot验证结果，qPCR和ELISA。本研究表明，PAL治疗是一种有效的肿瘤免疫治疗方法，扩大了肿瘤免疫治疗的范围。此外，这些发现为PAL的潜在临床应用奠定了理论基础。
