Gupta Parul, Singh Arpita, Verma Ajay Kumar, Kant Surya, Pandey Anuj Kumar, Khare Puneet, Prakash Ved
Department of Respiratory Medicine, King George's Medical University, Lucknow 226003, Uttar Pradesh, India.
Department of Pharmacology, Dr. Ram Manohar Lohia, Institute of Medical Sciences, Lucknow 226010, Uttar Pradesh, India.
Vaccines (Basel). 2022 Oct 26;10(11):1801. doi: 10.3390/vaccines10111801.
Taxanes, microtubule stabilizing agents, are extensively used in the treatment of non-small cell lung cancer (NSCLC). However, their clinical effectiveness remains restricted owing to significant adverse effects and drug resistance. Nanotechnology may guide chemotherapeutic drugs directly and selectively to malignant cells, improving their therapeutic efficiency. In the present study, we synthesized polylactic-co-glycolic acid (PLGA) based nanoparticles encapsulating docetaxel and evaluated their efficacy in non-small cell lung carcinoma (A549) cells and primary immune cells derived from humans. Docetaxel-PLGA nanoparticles (PLGA-Dtx) were synthesized and characterized using distinct methods. Moreover, the cytotoxicity of free docetaxel (Dtx) and Dtx-conjugated nanoparticles (PLGA-Dtx) was studied in A549 cells and peripheral blood mononuclear cells derived from humans. Furthermore, annexin V-FITC/PI staining was used to assess the mode of cell death. Additionally, human peripheral blood mononuclear cells (PBMCs) were used for assessing the associated immune response and cytokine profile following PLGA-Dtx treatment. Spherical PLGA-Dtx nanoparticles with a 150 ± 10 nm diameter and 70% encapsulation efficiency (EE) were synthesized. The MTT assay showed that the IC of PLGA-Dtx nanoparticles was significantly lower than free docetaxel in A549 cells. Cytotoxicity data also revealed the selective nature of PLGA-Dtx with no significant effects in normal human bronchial epithelial cells (BEAS-2B) and PBMCs derived from healthy donors. Interestingly, PLGA-Dtx exerted an improved effect and tempted both apoptosis and necroptosis, as evidenced by annexin V and propidium iodide-positive cells. Further, PLGA-Dtx-exposed A549 cells showed increased Cas-3, Cas-9, RIP-1, and RIP-3, indicating apoptosis and necroptosis. An increased pro-inflammatory response manifested from the enhancement of IFN-γ and TNF-α in PLGA-Dtx-exposed PBMCs, posed by the occurrence of necroptosis and the immune stimulatory effect of PLGA-Dtx. In conclusion, PLGA-Dtx has a selective anticancer potential and better immunostimulatory potential. Therefore, PLGA-Dtx may be useful for the treatment of non-small cell lung carcinoma.
紫杉烷类作为微管稳定剂，被广泛用于治疗非小细胞肺癌（NSCLC）。然而，由于显著的不良反应和耐药性，它们的临床疗效仍然受限。纳米技术可以将化疗药物直接且选择性地导向恶性细胞，提高其治疗效率。在本研究中，我们合成了包裹多西他赛的聚乳酸-羟基乙酸共聚物（PLGA）纳米颗粒，并评估了它们对非小细胞肺癌（A549）细胞和源自人类的原代免疫细胞的疗效。采用不同方法合成并表征了多西他赛-PLGA纳米颗粒（PLGA-Dtx）。此外，研究了游离多西他赛（Dtx）和Dtx偶联纳米颗粒（PLGA-Dtx）对A549细胞和源自人类的外周血单个核细胞的细胞毒性。此外，采用膜联蛋白V-FITC/PI染色评估细胞死亡方式。另外，使用人类外周血单个核细胞（PBMCs）评估PLGA-Dtx处理后的相关免疫反应和细胞因子谱。合成了直径为150±10nm、包封率（EE）为70%的球形PLGA-Dtx纳米颗粒。MTT试验表明，PLGA-Dtx纳米颗粒在A549细胞中的半数抑制浓度（IC）显著低于游离多西他赛。细胞毒性数据还揭示了PLGA-Dtx的选择性，对正常人支气管上皮细胞（BEAS-2B）和来自健康供体的PBMCs无显著影响。有趣的是，PLGA-Dtx发挥了更好的作用，诱导了凋亡和坏死性凋亡，膜联蛋白V和碘化丙啶阳性细胞证明了这一点。此外，暴露于PLGA-Dtx的A549细胞中，半胱天冬酶-3（Cas-3）、半胱天冬酶-9（Cas-9）、受体相互作用蛋白1（RIP-1）和受体相互作用蛋白3（RIP-3）增加，表明发生了凋亡和坏死性凋亡。坏死性凋亡的发生以及PLGA-Dtx的免疫刺激作用导致暴露于PLGA-Dtx的PBMCs中γ干扰素（IFN-γ）和肿瘤坏死因子-α（TNF-α）增强，表现出促炎反应增强。总之，PLGA-Dtx具有选择性抗癌潜力和更好的免疫刺激潜力。因此，PLGA-Dtx可能对非小细胞肺癌的治疗有用。
