突变体p53能够通过内源性的PDGFRb促进胰腺癌的转移PPT.ppt

Journal: Cell Reporter: XuWang IF: 33.116 Abstract Mutant p53 metastasis antiproliferative prometastatic phenotype PDGFRb p73/NF-Y metastasis invasion vitro vivo × prognostic marker Background Background 75% p53 mutant highly metastatic poor prognosis drug resistance Background KPC cell :stablely express small haipin RNA,lost remaning p53 wide-type alle KPflC cell:a p53 null cell line KPC mice: develop highly metastatic pancreatic cancer that faithfully mimics the human disease Methods 1.Wound Healing and Invasion Assays 2.RNA Sequencing and Data Analysis 3.Immunostaining and Microscopy 4.qRT-PCR 5.PDGFRb Luciferase Reporter Assay 6.Coimmunoprecipitation and Chromatin Immunoprecipitation 7.Immunohistochemistry and Immunofluorescence 8.Mouse Studies Question whether mutant p53 is needed to sustain the metastatic phenotype and how it is regulated？ RESULTS KPC+sh.Ctrl cell express mutant p53 划痕愈合率 侵袭细胞数 The invasiveness depend on mutant p53. Result 1：Sustained Expression of Mutant p53 Is Required for the Invasive Phenotype of Pancreatic Cancer Cells 转移瘤数量 whether mutant p53 expression was required to sustain the metastatic potential of KPC cells? 转移瘤荧光体视成像 Summary 1 these results demonstrate that mutant p53 can contribute to PDAC invasion and metastasis and that inhibiting its activity can have an antimetastatic effect Question how mutant p53 mediates the invasive phenotype of PDAC? mutant p53 can affect the invasive phenotype of pancreatic cancer cells RNA 测序，基因变化 IPA分析 Result 2：Transcriptional Profiling and Functional Screening Identify PDGFRb as a Downstream Mediator of Mutant p53 in Murine Pancreatic Cancer 1：SLC40A1 2：SNED1 3：PDGFRb hypothesis：PDGFRb could affect cell invasion PDGFRb转录水平 蛋白表达 mutant p53敲除 细胞增殖 不同亚型对侵袭的影响 不同亚型蛋白表达情况 红色荧光蛋白 绿色荧光蛋白 两种细胞比值 increased PDGFRb did not confer a selective advantage to tumor cell proliferation Summary 2 PDGFRb is not required for the proliferation and tumorigenic potential of p53 mutant murine cancer cells bu