Wang Wei, Suga Tadaharu, Hagimori Masayori, Kuroda Naotaka, Fuchigami Yuki, Kawakami Shigeru
Department of Pharmaceutical Informatics, Graduate School of Biomedical Sciences, Nagasaki University.
Department of Analytical Chemistry for Pharmaceutics, Graduate School of Biomedical Sciences, Nagasaki University.
Biol Pharm Bull. 2018;41(9):1448-1455. doi: 10.1248/bpb.b18-00335.
Oligoarginines (Rn) are becoming promising tools for the intracellular delivery of biologically active molecules. NuBCP-9, a peptide that induces apoptosis in B-cell lymphoma 2 (Bcl-2)-expressing cancer cells, has been reported to promote the uptake and non-specific cytotoxicity of R8, also called octaarginine. However, it is unknown whether a similar synergistic effect can be seen with other Rn. In this study, we conjugated NuBCP-9 with various Rn (n=8, 10, 12, 14) to investigate and compare their cellular uptake characteristics. In addition, their non-specific cytotoxicity and apoptosis-inducing abilities were evaluated. We found that NuBCP-9 conjugated with Rn enhanced cellular uptake mainly through clathrin-mediated endocytosis and macropinocytosis, and that the uptake pathways were not different from those used by unconjugated Rn. However, the cytotoxicity study showed that NuBCP-9-R12 and NuBCP-9-R14 conjugates enhanced non-specific cytotoxicity. We found that NuBCP-9-R10 conjugate had the highest uptake efficiency and induced correspondingly high levels of apoptosis, while resulting in a tolerable degree of non-specific toxicity.
寡聚精氨酸（Rn）正成为用于生物活性分子细胞内递送的有前景的工具。NuBCP-9是一种能在表达B细胞淋巴瘤2（Bcl-2）的癌细胞中诱导凋亡的肽，据报道它能促进R8（也称为八聚精氨酸）的摄取和非特异性细胞毒性。然而，尚不清楚其他Rn是否也能观察到类似的协同效应。在本研究中，我们将NuBCP-9与各种Rn（n = 8、10、12、14）偶联，以研究和比较它们的细胞摄取特性。此外，还评估了它们的非特异性细胞毒性和诱导凋亡的能力。我们发现，与Rn偶联的NuBCP-9主要通过网格蛋白介导的内吞作用和巨胞饮作用增强细胞摄取，并且摄取途径与未偶联的Rn所使用的途径没有差异。然而，细胞毒性研究表明，NuBCP-9-R12和NuBCP-9-R14偶联物增强了非特异性细胞毒性。我们发现，NuBCP-9-R10偶联物具有最高的摄取效率，并相应地诱导了高水平的凋亡，同时产生了可耐受程度的非特异性毒性。
