Immunogenetic effects of low dose (CEM43 30) magnetic nanoparticle hyperthermia and radiation in melanoma cells
Immunogenetic effects of low dose (CEM43 30) magnetic nanoparticle hyperthermia and radiation in melanoma cells
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DOI：
10.1080/02656736.2019.1627433
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发表时间：
2019-11-29
影响因子：
3.1
通讯作者：
Hoopes, P. Jack
中科院分区：
文献类型：
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作者：
Duval, Kayla E. A.;Vernice, Nicholas A.;Hoopes, P. Jack
目的：在该体外研究中，我们使用了RNA定量技术、nanoString和常规蛋白质分析技术（Western印迹）来评估B16鼠黑色素瘤细胞在中等磁性纳米颗粒热疗（mNPH）剂量（相当于在43摄氏度下30分钟（CEM 43 30）和/或临床相关的8戈伊辐射剂量）后的遗传和蛋白质表达。研究方法：将具有mNP（2.5 μ g Fe/106个细胞）的黑素瘤细胞沉淀并暴露于交变磁场（AMF）以产生靶向热剂量。热剂量由光纤探头精确监测，并自动保持在CEM 43 - 30。处理后24小时收获所有细胞。结果如下：mNPH剂量显示耐热性/免疫原性HSP 70基因和许多化学引诱物和toll样受体基因途径显著升高。8戈伊剂量也上调了一些重要的免疫和细胞毒性遗传和蛋白质途径。然而，mNPH/辐射组合是多种免疫和细胞毒性基因的最有效刺激物，包括HSP 70、癌症调节趋化因子CXCL 10、CXCL 11、T细胞运输趋化因子CXCR 3、先天免疫激活剂TLR 3、TLR 4、MDM 2和p53的mTOR负调节剂、促凋亡蛋白G41 A和细胞死亡受体Fas。重要的是，许多遗传变化通过蛋白质表达变化得到了准确验证，即，HSP 70、p-mTOR、p-MDM 2。结论：这些结果不仅表明，低剂量的mNPH和辐射独立地增加重要的免疫和细胞毒性基因的表达，而且当它们组合使用时，效果大大增强。
Objective: In this in vitro study we have used an RNA quantification technique, nanoString, and a conventional protein analysis technique (Western Blot) to assess the genetic and protein expression of B16 murine melanoma cells following a modest magnetic nanoparticle hyperthermia (mNPH) dose equivalent to 30 minutes @ 43 degrees C (CEM43 30) and/or a clinically relevant 8 Gy radiation dose. Methods: Melanoma cells with mNPs(2.5 mu g Fe/106 cells) were pelleted and exposed to an alternating magnetic field (AMF) to generate the targeted thermal dose. Thermal dose was accurately monitored by a fiber optic probe and automatically maintained at CEM43 30. All cells were harvested 24 hours after treatment. Results: The mNPH dose demonstrated notable elevations in the thermotolerance/immunogenic HSP70 gene and a number of chemoattractant and toll-like receptor gene pathways. The 8 Gy dose also upregulated a number of important immune and cytotoxic genetic and protein pathways. However, the mNPH/radiation combination was the most effective stimulator of a wide variety of immune and cytotoxic genes including HSP70, cancer regulating chemokines CXCL10, CXCL11, the T-cell trafficking chemokine CXCR3, innate immune activators TLR3, TLR4, the MDM2 and mTOR negative regulator of p53, the pro-apoptotic protein PUMA, and the cell death receptor Fas. Importantly a number of the genetic changes were accurately validated by protein expression changes, i.e., HSP70, p-mTOR, p-MDM2. Conclusion: These results not only show that low dose mNPH and radiation independently increase the expression of important immune and cytotoxic genes but that the effect is greatly enhanced when they are used in combination.