Cell-penetrating D-Isomer Peptides of p53 C-terminus: Long-term Inhibitory Effect on the Growth of Bladder Cancer
Cell-penetrating D-Isomer Peptides of p53 C-terminus: Long-term Inhibitory Effect on the Growth of Bladder Cancer
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DOI：
10.1016/j.urology.2009.10.002
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发表时间：
2010-04-01
期刊：
影响因子：
2.1
通讯作者：
Tomizawa, Kazuhito
中科院分区：
文献类型：
--
作者：
Araki, Daiji;Takayama, Kentaro;Tomizawa, Kazuhito
研究p53 C-末端的膜可渗透D-异构体与流感病毒血凝素-2蛋白（riHA 2）的NH 2-末端20-氨基酸肽的反向-反转形式连接的单次应用是否抑制膀胱癌细胞的生长。使用聚精氨酸转导p53可用于靶向和抑制膀胱癌细胞的生长。然而，该蛋白的细胞内半衰期短，重复应用是必要的，以实现抗肿瘤effect.Methods的p53羧基端肽共价偶联细胞穿透肽与D-或L-氨基酸合成。此外，肽通过二硫桥与riHA 2连接。将人膀胱癌细胞系与每种肽一起孵育，并使用WST测定法评估细胞活力。通过Hoechst和活性半胱氨酸蛋白酶-3染色证实凋亡细胞。将p53肽注射到J82细胞移植的重症联合免疫缺陷病小鼠中，以研究其对膀胱肿瘤的抗肿瘤作用。结果p53 C端与riHA 2连接的穿膜肽（d11 R-p53 C '-riHA 2和dFHV-p53 C'-riHA 2）可抑制膀胱癌细胞的生长并诱导其凋亡。结论p53 C末端D-异构体肽与riHA 2联合应用，可能成为治疗膀胱癌的一种新方法。泌尿学75：813-819，2010年。(C)2010年爱思唯尔公司
OBJECTIVES 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 NH2-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.METHODS 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.RESULTS 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%.CONCLUSIONS Peptide transduction therapy using the D- isomer p53 C-terminal peptide with riHA2 may be an innovative method for the treatment of bladder cancer. UROLOGY 75: 813-819, 2010. (C) 2010 Elsevier Inc.