Department of Physiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan.
Urology. 2010 Apr;75(4):813-9. doi: 10.1016/j.urology.2009.10.002. Epub 2009 Dec 6.
To investigate whether a single application of the membrane-permeable D-isomer of the p53 C-terminus connected with a retro-inverso version of the NH(2)-terminal 20-amino acid peptide of the influenza virus hemagglutinin-2 protein (riHA2) inhibited the growth of bladder cancer cells. The transduction of p53 using poly-arginine is useful for targeting and suppressing the growth of bladder cancer cells. However, the protein's intracellular half-life is short, and repeated application is necessary to achieve an anti-tumor effect.
The p53 carboxyl-terminal peptides covalently coupled with cell-penetrating peptides were synthesized with D- or L-amino acids. Moreover, the peptides were connected with riHA2 by a disulfide bridge. Human bladder cancer cell lines were incubated with each peptide and cell viability was assessed with the WST assay. Apoptotic cells were confirmed by Hoechst and active capase-3 staining. The p53 peptides were injected into severe combined immunodeficiency disease mice transplanted with J82 cells to investigate their anti-tumor effect on bladder tumors. A survival curve was plotted using the Kaplan-Meier method.
A single application of cell-penetrating D-isomer peptides of the p53 C-terminus connected with riHA2 (d11R-p53C'-riHA2 and dFHV-p53C'-riHA2) inhibited the growth and induced the apoptosis of bladder cancer cells. The tumor-bearing mice treated only with vehicle had a mean survival time of 12 days, whereas treatment with d11R-p53C'-riHA2 resulted in a long-term survival rate of 50%.
Peptide transduction therapy using the D-isomer p53 C-terminal peptide with riHA2 may be an innovative method for the treatment of bladder cancer.
研究与流感病毒血凝素-2 蛋白的 NH2-末端 20 个氨基酸肽的反向倒转版本连接的膜通透性 D-异构体 p53 C 末端是否能抑制膀胱癌细胞的生长。使用多聚精氨酸转导 p53 可用于靶向和抑制膀胱癌细胞的生长。然而，该蛋白质的细胞内半衰期较短，需要重复应用才能达到抗肿瘤效果。
用 D-或 L-氨基酸合成共价连接细胞穿透肽的 p53 羧基末端肽，并且通过二硫键将肽与 riHA2 连接。用每种肽孵育人膀胱癌细胞系，并用 WST 测定法评估细胞活力。通过 Hoechst 和活性 capase-3 染色确认凋亡细胞。将 p53 肽注入移植 J82 细胞的严重联合免疫缺陷疾病小鼠中，以研究其对膀胱癌的抗肿瘤作用。使用 Kaplan-Meier 方法绘制生存曲线。
单次应用与 riHA2 连接的 p53 C 末端细胞穿透 D-异构体肽（d11R-p53C'-riHA2 和 dFHV-p53C'-riHA2）抑制了膀胱癌细胞的生长并诱导了其凋亡。仅用载体治疗的荷瘤小鼠的平均存活时间为 12 天，而用 d11R-p53C'-riHA2 治疗则导致 50%的长期存活率。
使用与 riHA2 连接的 D-异构体 p53 C 末端肽的肽转导治疗可能是治疗膀胱癌的一种创新方法。
