南方医科大学学报

南方医科大学学报 ›› 2022, Vol. 42 ›› Issue (11): 1681-1688.doi: 10.12122/j.issn.1673-4254.2022.11.12

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类黑色素纳米颗粒联合近红外光照射具有良好的抗卵巢癌作用

杨洁容，陈小平   

  1. 广东省第二中医院妇科，广东 广州 510095
  • 出版日期:2022-11-20 发布日期:2022-11-30

Natural melanin-based nanoparticles with photothermal/photodynamic activities induce ovarian cancer cell death

YANG Jierong, CHEN Xiaoping   

  1. Department of Gynecology, Guangdong Second Hospital of Traditional Chinese Medicine, Guangzhou 510095, China
  • Online:2022-11-20 Published:2022-11-30

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摘要： 目的 探讨类黑色素（PDA）纳米的制备、物化表征及其与光热治疗（PTT）/光动力治疗（PDT）协同抗肿瘤效率。方法采用透射电镜（TEM），水合粒径分析及Zeta电位分析分析PDA纳米的形貌、粒径大小，利用近红外激发光（NIR）探究其光热及光动力性能。将卵巢癌细胞分别用全培（Control组）、单纯PDA、PDA+NIR（0.7 W/cm2与1.0 W/cm2）进行处理，进一步采用流式细胞术、细胞毒性试验（CCK-8）、Transwell实验与单侧皮下荷瘤模型评估类黑色素纳米的PTT/PDT协同抗肿瘤作用。结果 制备的 PDA 纳米为-20 mV及100 nm 左右的圆球颗粒。在 NIR（0.7 W/cm2与 1.0 W/cm2）照射下其变化温度值（ΔT）分别约为15 ℃或30 ℃，同时可产生大量单线态氧，具有优良的PTT/PDT性能。流式细胞学结果表明：相比于空白组，PDA纳米不会引起卵巢癌细胞活性氧生成及线粒体膜电位变化，PDA联合NIR照射可诱导卵巢癌细胞内活性氧显著上升以及线粒体膜电位下降（P<0.001）；且相比较于低功率NIR（0.7 W/cm2）光照组，高功率NIR（1.0 W/cm2）处理的细胞活性氧水平更高、线粒体膜电位下降越显著（P<0.01）。CCK8实验表明：相比于空白组，PDA对肿瘤细胞无明显毒性，PDA联合NIR照射对卵巢癌细胞具有显著杀伤作用（P<0.001），且相比较于低功率NIR（0.7 W/cm2）光照组，高功率NIR（1.0 W/cm2）处理的细胞存活率越低（P<0.001）。Transwell实验表明：相比于空白组，PDA不能抑制卵巢癌细胞的侵袭迁移能力，PDA联合NIR照射能显著抑制卵巢癌细胞的侵袭迁移能力（P<0.001），且相比较于低功率NIR（0.7 W/cm2）光照组，高功率NIR（1.0 W/cm2）处理的卵巢癌细胞侵袭迁移数目越少（P<0.001）。小鼠单侧皮下荷瘤模型表明：相比于生理盐水组和单独的PDA组，PDA联合NIR照射能发挥显著的体内抗肿瘤作用（P<0.001）。结论 类黑色素联合近红外光照射具有良好的抗肿瘤效应，是卵巢癌治疗的一种潜在新方法。

关键词: 类黑色素纳米；纳米药物；近红外光照射；卵巢癌

Abstract: Objective To investigate the physicochemical characteristics of natural melanin-like nanoparticles (PDA NPs) and their synergistic anti-tumor efficacy with photothermal and photodynamic (PTT/PDT) therapy. Methods The chemically synthesized PDA NPs were characterized using transmission electron microscope (TEM), dynamic light scattering (DLS) and Zeta potential analysis, and their photothermal and photodynamic properties were assessed with near-infrared excitation light (NIR). The antitumor efficacy of free PDA or PDA combined with NIR irradiation (0.7 and 1.0 W/cm2) was evaluated in ovarian cancer cells using flow cytometry, Cell Counting Kit-8 (CCK-8), and Transwell assay and in a mouse model bearing subcutaneous ovarian cancer xenograft. Results The synthesized PDA NPs showed a spherical morphology with diameters around 100 nm and a zeta potential of -20 mV. Upon NIR irradiation at 0.7 and 1.0 W/cm2, the particles underwent temperature changes (ΔT) of about 15 and 30 ℃, respectively, and produced a large amount of singlet oxygen, demonstrating their excellent PTT/PDT properties. In ovarian cancer cells, treatment with PDA NPs alone did not induce obvious changes in reactive oxygen species (ROS) production or mitochondrial membrane potential (MTP), but when combined with NIR irradiation, these particles caused a significant increase of ROS and a reduction of MTP (P<0.001), and such changes were more prominent with high power NIR (P<0.01). PDA NPs alone showed no obvious cytotoxicity, but the combination of PDA with NIR irradiation produced potent killing effect on ovarian cancer cells (P<0.001), and the effect was much stronger with a high power irradiation (P<0.001). While PDA alone showed no inhibitory effect on tumor cell metastasis, the combined treatment with PDA and NIR irradiation, especially at a high power, significantly suppressed invasion and migration of ovarian cancer cells (P<0.001). In the tumor-bearing mouse model, the combined treatment, but not PDA alone, produced a significant inhibitory effect on tumor growth (P<0.001). Conclusion PDA NPs combined with NIR has a strong anti-tumor effect, suggesting a potential new therapeutic strategy for ovarian cancer.

Key words: melanin-like nanoparticles; nanomedicine; mear-infrared light irradiation; ovarian cancer