Department of Biochemistry and Molecular Biomedicine, Facultat of Biologia, Universitat de Barcelona, Barcelona, Spain; Departamento de Biología, Facultad de Ciencias, Universidad Autónoma de Madrid, Madrid, Spain; Communication Department, Hospital Clinic of Barcelona, Barcelona, Spain.
Department of Biochemistry and Molecular Biomedicine, Facultat of Biologia, Universitat de Barcelona, Barcelona, Spain.
Eur J Pharm Sci. 2017 Nov 15;109:65-77. doi: 10.1016/j.ejps.2017.07.024. Epub 2017 Jul 19.
Irinotecan (CPT-11) is an effective chemotherapeutic agent widely used to treat different cancers. Otherwise, the liposomal delivery of anti-tumor agents has been shown to be a promising strategy. The aim of this study has been to analyze the effect of liposomal CPT-11 (CPT-11lip) on two human cell lines (Hs68 and HeLa) to establish the suitability of this CPT-11 nanocarrier. We have demonstrated the highest uptake of CPT-11lip in comparison with that of CPT-11sol, in lactate buffer, and that CPT-11lip was internalized in the cells through an endocytic process whereas CPT-11sol does so by passive diffusion. CPT-11lip was not cytotoxic to normal fibroblast Hs68 cells, but induced a massive apoptosis accompanied by cell senescence in HeLa cells. CPT-11lip treatment modified the morphology of HeLa cells, induced different cell cycle alterations and accumulated into lysosomes in both cell lines. In particular, CPT-11lip treatment showed that surviving HeLa cells remained in a state of senescence whereas only a temporal growth arrest was induced in Hs68 cells. Results of RT-PCR indicated that the different responses in Hs68 (survival) and HeLa cells (apoptotic death), seemed to be induced by a p53- and p53- independent mechanism, respectively. An analysis of DNA damage also determined that released CPT-11 from liposomes was able to reach the nucleus and exert a genotoxic effect in both cell lines, which was repaired in Hs68 but not in HeLa cells. All results indicate that phospholipid-cholesterol liposomes possess optimum properties for CPT-11 delivery, being biocompatible and selectively cytotoxic against HeLa tumorigenic cells.
伊立替康（CPT-11）是一种有效的化疗药物，广泛用于治疗不同的癌症。然而，抗肿瘤药物的脂质体递送已被证明是一种很有前途的策略。本研究旨在分析脂质体伊立替康（CPT-11lip）对两种人细胞系（Hs68 和 HeLa）的影响，以确定这种 CPT-11 纳米载体的适用性。我们已经证明，与 CPT-11sol 相比，CPT-11lip 在乳酸缓冲液中的摄取率更高，并且 CPT-11lip 通过内吞作用被细胞内化，而 CPT-11sol 则通过被动扩散被内化。CPT-11lip 对正常成纤维细胞 Hs68 细胞没有细胞毒性，但在 HeLa 细胞中诱导大量凋亡并伴有细胞衰老。CPT-11lip 处理改变了 HeLa 细胞的形态，诱导了两种细胞系中不同的细胞周期改变，并在溶酶体中积累。特别是，CPT-11lip 处理表明，存活的 HeLa 细胞仍然处于衰老状态，而 Hs68 细胞仅诱导暂时的生长停滞。RT-PCR 结果表明，Hs68（存活）和 HeLa 细胞（凋亡死亡）中的不同反应似乎分别由 p53 依赖和非依赖机制诱导。DNA 损伤分析也表明，从脂质体中释放的 CPT-11 能够到达细胞核，并在两种细胞系中产生遗传毒性作用，该作用在 Hs68 中得到修复，但在 HeLa 细胞中未得到修复。所有结果表明，磷脂胆固醇脂质体具有 CPT-11 递药的最佳特性，对 Hs68 细胞具有生物相容性和选择性细胞毒性，但对 HeLa 肿瘤细胞没有作用。
