The synthetic peptide P111-136 derived from the C-terminal domain of heparin affin regulatory peptide inhibits tumour growth of prostate cancer PC-3 cells
The synthetic peptide P111-136 derived from the C-terminal domain of heparin affin regulatory peptide inhibits tumour growth of prostate cancer PC-3 cells
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DOI：
10.1186/1471-2407-11-212
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发表时间：
2011-05-30
期刊：
影响因子：
3.8
通讯作者：
Delbe, Jean
中科院分区：
文献类型：
--
作者：
Hamma-Kourbali, Yamina;Bermek, Oya;Delbe, Jean
背景资料：肝素亲和调节肽（HARP），也称为多效生长因子，是一种肝素结合的分泌因子，其在几种肿瘤中过表达并与肿瘤生长、血管生成和转移相关。由氨基酸111 - 136组成的HARP的C-末端部分特别参与其生物活性，并且我们先前确定由相同氨基酸（P111-136）组成的合成肽能够抑制HARP的生物活性。在这里，我们评估P111-136的能力，以抑制在体外和体内的人肿瘤细胞系PC-3的生长，具有HARP自分泌loop.Methods：PC-3细胞的总裂解物与生物素化的P111-136孵育，并拉下来的HARP受体在Western印迹的存在。在体外，P111-136对PC-3细胞中HARP自分泌环的影响通过在软琼脂中的集落形成来确定。在体内，将PC-3细胞接种于无胸腺裸鼠的侧腹。动物接受P111-136（5 mg/kg/天）给药25天。在治疗期间评估肿瘤体积。动物处死后，通过免疫组织化学评价肿瘤细胞凋亡和相关的血管生成。结果：通过pull down实验，我们鉴定了HARP受体RPTPb/zeta、ALK和核仁素为P111-136结合蛋白。在体外，P111-136剂量依赖性地抑制PC-3细胞集落形成。用P111-136治疗显著抑制异种移植模型中的PC-3肿瘤生长以及肿瘤血管生成。结论：P111 - 136能明显抑制HARP对PC-3细胞生长的促有丝分裂作用。这种抑制可能与该肽与HARP受体（ALK、RPTPb/zeta、核仁素）的直接或间接结合有关。在体内，P111-136治疗显著抑制PC-3肿瘤生长和相关的血管生成。因此，P111-136可能被认为是一种有趣的药理学工具，可以干扰肿瘤生长，目前正在其他癌症类型中进行评估。
Background: Heparin affin regulatory peptide (HARP), also called pleiotrophin, is a heparin-binding, secreted factor that is overexpressed in several tumours and associated to tumour growth, angiogenesis and metastasis. The C-terminus part of HARP composed of amino acids 111 to 136 is particularly involved in its biological activities and we previously established that a synthetic peptide composed of the same amino acids (P111-136) was capable of inhibiting the biological activities of HARP. Here we evaluate the ability of P111-136 to inhibit in vitro and in vivo the growth of a human tumour cell line PC-3 which possess an HARP autocrine loop.Methods: A total lysate of PC-3 cells was incubated with biotinylated P111-136 and pulled down for the presence of the HARP receptors in Western blot. In vitro, the P111-136 effect on HARP autocrine loop in PC-3 cells was determined by colony formation in soft agar. In vivo, PC-3 cells were inoculated in the flank of athymic nude mice. Animals were treated with P111-136 (5 mg/kg/day) for 25 days. Tumour volume was evaluated during the treatment. After the animal sacrifice, the tumour apoptosis and associated angiogenesis were evaluated by immunohistochemistry. In vivo anti-angiogenic effect was confirmed using a mouse Matrigel T plug assay.Results: Using pull down experiments, we identified the HARP receptors RPTPb/zeta, ALK and nucleolin as P111-136 binding proteins. In vitro, P111-136 inhibits dose-dependently PC-3 cell colony formation. Treatment with P111-136 inhibits significantly the PC-3 tumour growth in the xenograft model as well as tumour angiogenesis. The angiostatic effect of P111-136 on HARP was also confirmed using an in vivo Matrigel T plug assay in miceConclusions: Our results demonstrate that P111-136 strongly inhibits the mitogenic effect of HARP on in vitro and in vivo growth of PC-3 cells. This inhibition could be linked to a direct or indirect binding of this peptide to the HARP receptors (ALK, RPTPb/zeta, nucleolin). In vivo, the P111-136 treatment significantly inhibits both the PC-3 tumour growth and the associated angiogenesis. Thus, P111-136 may be considered as an interesting pharmacological tool to interfere with tumour growth that has now to be evaluated in other cancer types.