基于PNRC的抗Ras PDT融合多肽通过影响核受体途径抑制MCF-7细胞的增殖  

Anti-proliferative Effect of PDT-PARP anti-Ras peptidimer on MCF-7 Cells Via Interfering Nuclear Receptor Signaling

作　　者：王越[1] 陈彬[1] 李渝萍[1] 陈健[1] 娄桂予[1] 张艳[1] 何凤田[1] 周度金[1] 

机构地区：[1]第三军医大学生物化学与分子生物学教研室,重庆市400038

出　　处：《医学分子生物学杂志》2008年第6期471-475,共5页Journal of Medical Molecular Biology

基　　金：国家自然科学基金(No.30672055);重庆市自然科学基金(No.CSTC.2006BB5352)~~

摘　　要：目的研究设计、合成的基于富含脯氨酸核受体辅活化子(proline-rich nuclear receptor coactivator protein,PNRC)的抗RasPDT融合多肽(PDT-PARP)在MCF-7乳腺癌细胞中的定位、对核受体信号传导途径的影响及其抗肿瘤细胞增殖活性。方法用多肽合成仪合成PNRC的抗RasPDT融合多肽PDT-PARP、不含PDT的PARP、单独的PDT多肽以及FITC标记的PDT-PARP、PARP,HPLC分析和纯化后,荧光显微镜下观察PDT-PARP、PARP在MCF-7细胞中的分布情况,流式细胞仪做定量分析。虫荧光素酶报告基因检测PDT-PARP对野生型PNRC辅活化ER反式激活功能的影响。MTS检测PDT-PARP对MCF-7细胞的抗增殖活性。结果PDT-PARP能进入MCF-7细胞;该多肽能抑制野生型PNRC对ER反式激活功能的辅活化作用,并对MCF-7细胞的增殖具有一定的抑制作用。结论PNRC抗RasPDT融合多肽可通过影响核受体途径抑制乳腺癌细胞的增殖。Objective To study the localization, the effect on nuclear receptor signaling and the anti-proliferation activity of the PNRC based and PDT fused anti-Ras peptidimer, PDT-PARP, in breast cancer cells MCF-7. Methods Five polypeptides were synthesized b v peptide synthesizer, including PDT-PARP, PARP alone, PDT alone and fluorescence labeled FITC-PDT-PARP and FITC-PARP. The polypeptides were analysized and purified by high performance liquid chromatography （HPLC） . The localization of FITC-PDT-PARP and FITC-PARP in MCF-7 was observed by fluorescence microscopy and FACS. The effect of PDT-PARP on wild type PNRC to co-activate the trans-activity of the nuclear estrogen receptor ER was evaluated by luciferase reporter assay. The anti-proliferation activity of PDT-PNRC on MCF-7 cells was detected by MTS. Results PDT-PARP, but not PARP alone, entered into MCF-7 cells and inhibited coactivition of wild type PNRC on the trans-activity of ER. PDT-PARP inhibited the proliferation of MCF-7 in a dose-dependent man- ner. Conclusion The PNRC based and PDT fused anti-ras peptidimer PDT-PARP inhibits the proliferation of MCF-7 cells by interfering nuclear receptor ER signaling.

关 键 词：PNRC 融合多肽 核受体 信号传导 

分 类 号：R737.31[医药卫生—肿瘤]