College of Pharmacy, Keimyung University, Daegu 42601, South Korea.
College of Pharmacy, Yeungnam University, Gyeongsan 38541, Gyeongbuk, South Korea.
Mol Pharm. 2020 Nov 2;17(11):4386-4400. doi: 10.1021/acs.molpharmaceut.0c00856. Epub 2020 Oct 20.
The mechanism of cell death has attracted a great deal of research interest in the design of antitumor therapy in recent days. Several attempts have been carried out in this direction and in our study also, we studied this phenomenon with the design of panitumumab (PmAb)-conjugated and temozolomide (TMZ)-loaded poly(lactic--glycolic acid) nanoparticles (PLGA-NPs), termed PmAb-TMZ-PLGA-NPs. First, PmAb was functionalized on the surface of TMZ-PLGA-NPs using ethyl(dimethylaminopropyl)carbodiimide (EDC)--hydroxysuccinimide (NHS) chemistry. Targeted PLGA-NPs significantly enhanced the cellular uptake of nanoparticles in the U-87 MG cell line as a result of the high epidermal growth factor receptor (EGFR) expression, compared to the LN229 cell line. Our study demonstrated that following the treatment of PmAb-TMZ-PLGA-NPs, a more pronounced anticancer effect was noticed in comparison with free TMZ and TMZ-PLGA-NPs. Further, a more pronounced cytotoxic effect of PmAb-TMZ-PLGA-NPs was observed in the high EGFR-overexpressed glioblastoma multiforme (GBM) model (U-87 MG) cell line compared to the low EGFR GBM model (LN229). Our study demonstrated that the treatment of PmAb-TMZ-PLGA-NPs in GBM tried to adopt the autophagic pathway of the cell survival mechanism with the elevated level of autophagic marker (Beclin-1 and LC3B) at 24 h time point, thereby suppressing the expression of caspase-9 and PARP. However, at the 48 h time point, the elevated expression of caspase-9 and PARP with the downregulation of Beclin-1 and LC3B, following the treatment of PmAb-TMZ-PLGA-NPs in the GBM model, suggested that apoptotic cell death was superior over autophagic cell survival. It was also noteworthy the activation of caspase-9 was correlated with the continuous overproduction of reactive oxygen species up to a 48 h time point after the treatment of PmAb-TMZ-PLGA-NPs. This result sheds light on the biological effect of targeted chemotherapy and illustrates that PmAb-TMZ-PLGA-NPs could be applied for EGFR-overexpressed different cancer models.
细胞死亡的机制在抗肿瘤治疗的设计中引起了广泛的研究兴趣。在这方面已经进行了一些尝试，在我们的研究中，我们还研究了这种现象，设计了帕尼单抗（PmAb）偶联和替莫唑胺（TMZ）负载的聚乳酸-羟基乙酸共聚物纳米粒子（PLGA-NPs），称为 PmAb-TMZ-PLGA-NPs。首先，通过使用 1-乙基-3-（3-二甲氨基丙基）碳二亚胺（EDC）-N-羟基琥珀酰亚胺（NHS）化学，将 PmAb 功能化到 TMZ-PLGA-NPs 表面。与 LN229 细胞系相比，由于高表皮生长因子受体（EGFR）表达，靶向 PLGA-NPs 显著增强了 U-87 MG 细胞系中纳米颗粒的细胞摄取。我们的研究表明，与游离 TMZ 和 TMZ-PLGA-NPs 相比，在用 PmAb-TMZ-PLGA-NPs 处理后，观察到更明显的抗癌作用。此外，与低 EGFR 胶质母细胞瘤模型（LN229）相比，在高 EGFR 过表达的多形性胶质母细胞瘤（GBM）模型（U-87 MG）细胞系中观察到 PmAb-TMZ-PLGA-NPs 的更明显的细胞毒性作用。我们的研究表明，在 GBM 中用 PmAb-TMZ-PLGA-NPs 治疗试图通过自噬途径来适应细胞生存机制，在 24 小时时间点上调自噬标记物（Beclin-1 和 LC3B），从而抑制半胱天冬酶-9 和 PARP 的表达。然而，在 48 小时时间点，在用 PmAb-TMZ-PLGA-NPs 处理 GBM 模型后，半胱天冬酶-9 和 PARP 的表达升高，Beclin-1 和 LC3B 的表达下调，表明细胞凋亡死亡优于自噬细胞存活。值得注意的是，在用 PmAb-TMZ-PLGA-NPs 处理后，直到 48 小时时间点，半胱天冬酶-9 的激活与活性氧的持续过度产生相关。这一结果揭示了靶向化疗的生物学效应，并表明 PmAb-TMZ-PLGA-NPs 可应用于 EGFR 过表达的不同癌症模型。
