Institute of Molecular Biosciences, University of Graz, Humboldtstraße 50/III, A-8010 Graz, Austria; Innovative Cancer Models, St. Anna Children's Cancer Research Institute, Zimmermannplatz 10, A-1090 Wien, Austria; BioTechMed-Graz, Austria.
Institute of Molecular Biosciences, University of Graz, Humboldtstraße 50/III, A-8010 Graz, Austria; BioTechMed-Graz, Austria; Center for Medical Research, Medical University of Graz, Stiftingtalstraße 24, A-8010 Graz, Austria.
Biochim Biophys Acta Biomembr. 2020 Aug 1;1862(8):183264. doi: 10.1016/j.bbamem.2020.183264. Epub 2020 Mar 7.
The aim of this study was to develop effective and specific anti-cancer drugs based on membrane active peptides. In previous studies we showed that human lactoferricin (hLFcin) derived peptides facilitate specific killing of cancer cells. These antitumor peptides were found by conventional melanoma two-dimensional (2D) cell cultures to induce apoptosis of cancer cells and to specifically target lipid phosphatidylserine located on the outside of cancer cell membranes. In order to have a more relevant in vitro model able to mimic the natural microenvironments of tumor tissues we established three-dimensional (3D) multicellular tumor spheroids (MCTS). We used a set of (retro) di-peptides derived from LF11, an 11 amino acid long fragment of hLFcin, which differed in peptide length, positive net charge and hydrophobicity and determined antitumor activity and non-specific toxicity on non-neoplastic cells using 2D and 3D model systems. 2D studies unveiled a correlation between length, positive net charge and hydrophobicity of peptides and their specific antitumor activity. (Retro) di-peptides as R-DIM-P-LF11-215 and DIM-LF11-322 with a net charge of +9 and moderate hydrophobicity exhibited the highest specific antitumor activity. Further evaluation of the peptides anticancer activity by 3D in vitro studies confirmed their higher activity and cancer specificity compared to their parent R-DIM-P-LF11, with the exception of DIM-LF11-339. This highly hydrophobic peptide caused cell death mainly at the border of tumor spheroids indicating that too high hydrophobicity may prevent peptides from reaching the center of the spheroids.
本研究旨在基于膜活性肽开发有效且特异的抗癌药物。在先前的研究中，我们发现人乳铁蛋白衍生肽（hLFcin）衍生肽能够促进癌细胞的特异性杀伤。这些抗肿瘤肽在传统的黑色素瘤二维（2D）细胞培养中被发现，可诱导癌细胞凋亡，并特异性靶向位于癌细胞膜外的脂质磷脂酰丝氨酸。为了建立更相关的体外模型，能够模拟肿瘤组织的天然微环境，我们建立了三维（3D）多细胞肿瘤球体（MCTS）。我们使用了一组（反式）二肽，这些二肽源自 LF11，是 hLFcin 的一个 11 个氨基酸长的片段，其在肽长度、净正电荷和疏水性方面存在差异，并使用 2D 和 3D 模型系统确定了对非肿瘤细胞的抗肿瘤活性和非特异性毒性。2D 研究揭示了肽的长度、净正电荷和疏水性与其特异性抗肿瘤活性之间的相关性。（反式）二肽 R-DIM-P-LF11-215 和 DIM-LF11-322 具有+9 的净正电荷和中等疏水性，表现出最高的特异性抗肿瘤活性。通过 3D 体外研究进一步评估肽的抗癌活性，证实与母体 R-DIM-P-LF11 相比，其活性和抗癌特异性更高，除了 DIM-LF11-339。这种高度疏水性的肽主要在肿瘤球体的边界处引起细胞死亡，表明过高的疏水性可能会阻止肽到达球体的中心。
