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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 investigate the localization of PDT-PARP (anti-RasPDT fusion polypeptide) based on the proline-rich nuclear receptor coactivator protein (PNRC) in MCF-7 breast cancer cells. On nuclear receptor signal transduction pathway and its anti-tumor cell proliferative activity. METHODS: PNRC anti-RasPDT fusion polypeptide PDT-PARP, PDT-free PARP, PDT polypeptide and FITC-labeled PDT-PARP were synthesized by peptide synthesizer. PARP was analyzed by HPLC and purified by fluorescence microscope. PARP distribution in MCF-7 cells, quantitative analysis by flow cytometry. Effect of luciferase reporter gene on the transactivation of wild-type PNRC co-activated ER by luciferase reporter assay. MTS detection of PDT-PARP on MCF-7 cells antiproliferative activity. Results PDT-PARP could enter into MCF-7 cells. This peptide could inhibit the activation of ER transactivation by wild-type PNRC and inhibit the proliferation of MCF-7 cells. Conclusion PNRC anti-RasPDT fusion polypeptide can inhibit the proliferation of breast cancer cells by affecting the nuclear receptor pathway.