目的: 探讨人参炔醇对巨噬细胞表型和功能的调控作用及对乳腺癌细胞生物学行为的影响。方法: 用人参炔醇处理4T1乳腺癌细胞，观察各项生物学行为变化；用4T1乳腺癌细胞培养上清液预处理RAW264.7 巨噬细胞系48 h，诱导其向M2型巨噬细胞极化；极化后的巨噬细胞再分别经不同浓度人参炔醇(0，8，16 μg/mL)处理24 h，用qRTPCR、蛋白质印迹、流式细胞术、ELISA法联合检测各组表型、抗原递呈相关分子MHCⅡ类分子、CD80、CD86表达改变、分泌谱变化，以及细胞系迁移、侵袭等。结果： 与0 μg/mL组相比，乳腺癌细胞系经8，16 μg/mL人参炔醇处理后，细胞增殖率明显降低(P均＜0.01)，细胞凋亡率明显增加(P均＜0.05)，癌细胞趋化作用减弱； M2型巨噬细胞经16 μg/mL人参炔醇刺激后，细胞表型逆转，Arg1表达明显降低(P均＜0.01)，iNOS表达明显增加(P均＜0.01)，MHCⅡ、CD80、CD86表达升高；并伴随IL1β、TNFα表达升高(P均＜0.01)，IL4、IL8、IL10表达降低(P均＜0.05)；经人参炔醇处理的巨噬细胞能够明显减少乳腺癌细胞的迁移与侵袭。结论： 人参炔醇可重塑巨噬细胞表型和功能，从而间接改变乳腺癌细胞生物学行为。
Abstract
Objective: To investigate the effects of panaxynol on the phenotype and function of macrophages and the biological behavior of breast cancer cells. Methods: Firstly, breast cancer 4T1 cells were directly treated with panaxynol, and various biological behavior changes were observed. Secondly, the macrophage cell line RAW264.7 was stimulated with breast cancer cell 4T1 culture supernatant for 48 h to induce macrophage polarization to M2 macrophages, and then macrophages were treated with different concentrations of panaxynol(0, 8, 16 μg/mL) for 24 h. qRTPCR, Western blotting, flow cytometry and ELISA were used to detect the phenotypic and antigenpresenting related molecules expression changes, such as MHCⅡ, CD80 and CD86, and the secretion spectrum changes, and the influence of panaxynol on the migration and invasion of breast cancer cell lines. Results: Compared with the 0 μg/mL group, the breast cancer cells proliferation was inhibited by the treatment with 8, 16 μg/mL panaxynol(P＜0.01), the cell apoptosis rate was enhanced(P＜0.05), and the chemotaxis of cancer cells was weakened. In addition, panaxynolstimulated macrophage showed the M2 phenotype. After stimulation with panaxynol(16 μg/mL), the macrophage phenotype was reversed, the expression of Arg1 was decreased(P＜0.01), while the expression of iNOS was increased(P＜0.01), and the surface antigenpresenting molecules(MHCⅡ, CD80, CD86) was increased significantly, accompanied by the expression level of IL4, IL8, IL10 downregulated(P＜0.01) and IL1β, TNFα upregulated(all P＜0.05). In addition, panaxynolstimulated macrophages could significantly reduce the number of migrating and invading breast cancer cells. Conclusion: Panaxynol could remodel macrophage phenotype and function, and therefore regulate the biological behavior of breast cancer cells.
关键词
参炔醇 /
巨噬细胞 /
乳腺癌 /
极化 /
肿瘤细胞 /
细胞增殖 /
细胞凋亡
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参考文献
［1］Dong D, Zhang G, Yang J, et al. The role of iron metabolism in cancer therapy focusing on tumorassociated macrophages. J Cell Physiol, 2019，234(6):8028-8039.
［2］Cassetta L, Pollard JW. Targeting macrophages: therapeutic approaches in cancer. Nat Rev Drug Discov, 2018，17(12)：887-904.
［3］Yamaguchi T, Fushida S, Yamamoto Y, et al. Tumorassociated macrophages of the M2 phenotype contribute to progression in gastric cancer with peritoneal dissemination. Gastric Cancer, 2016, 19(4): 1052-1065.
［4］Medrek C, Ponten F, Jirstrom K, et al. The presence of tumor associated macrophages in tumor stroma as a prognostic marker for breast cancer patients. BMC Cancer, 2012, 12: 306.
［5］Gupta V, Yull F, Khabele D. Bipolar tumorassociated macrophages in ovarian cancer as targets for therapy. Cancers (Basel), 2018, 10(10). pii: E366.
［6］Sharda DR, Yu S, Ray M, et al. Regulation of macrophage arginase expression and tumor growth by the Ron receptor tyrosine kinase. J Immunol, 2011, 187(5): 2181-2192.
［7］Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation. Cell, 2011,144(5): 646-674.
［8］Ho MY, Tang SJ, Chuang MJ, et al. TNFα induces epithelialmesenchymal transition of renal cell carcinoma cells via a GSK3βdependent mechanism. Mol Cancer Res, 2012, 10(8): 1109-1119.
［9］段贤春, 汪永忠, 居靖. 人参炔醇研究进展. 安徽医药, 2008, 12(1)：1-3.
［10］Le HT, Nguyen HT, Min HY, et al. Corrigendum to “Panaxynol, a natural Hsp90 inhibitor, effectively targets both lung cancer stem and nonstem cells”.. Cancer Lett, 2018, 424: 136.
［11］Yan Z, Yang R, Jiang Y, et al. Induction of apoptosis in human promyelocytic leukemia HL60 cells by panaxynol and panaxynol. Molecules, 2011, 16(7): 5561-5573.
［12］Kim JY, Lee K, Kim SH, et al. Inhibitory effect of tumor cell proliferation and induction of G2/M cell cycle arrest by panaxytriol. Planta Medica, 2002, 68(2):119-122.
［13］Sharda DR, Yu S, Ray M, et al. Regulation of macrophage arginase expression and tumor growth by the Ron receptor tyrosine kinase. J Immunol, 2011, 187(5):2181-2192.
［14］Ding M, Fu X, Tan H, et al. The effect of vascular endothelial growth factor C expression in tumorassociated macrophages on lymphangiogenesis and lymphatic metastasis in breast cancer. Mol Med Rep, 2012, 6(5):1023-1029.
［15］Afik R, Zigmond E, Vugman M, et al. Tumor macrophages are pivotal constructors of tumor collagenous matrix. J Exp Med, 2016, 213(11): 2315-2331.
［16］Ashkar AA, Gillgrass A, Krneta T. The influence of macrophages and the tumor microenvironment on natural killer cells. Curr Mol Med, 2013, 13(1)：68-79.
［17］Huang Z, Yang Y, Jiang Y, et al. Antitumor immune responses of tumorassociated macrophages via tolllike receptor 4 triggered by cationic polymers. Biomaterials, 2013, 34(3):746-755.
［18］Yang H, Kim C, Kim MJ, et al. Soluble vascular endothelial growth factor receptor3 suppresses lymphangiogenesis and lymphatic metastasis in bladder cancer. Mol Cancer, 2011, 10: 36.
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脚注
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基金
江苏省社会发展项目（BE2016716）；江苏省高校重点实验项目（16KJA320005）
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