突变体p53能够通过内源性的PDGFRb促进胰腺癌的转移教程文件.ppt

肺转移 肺组织病理切片 Result 5： Modulation of PDGFRb Expression Levels Mediates the Phenotypic Effects of Mutant p53 Depletion In Vivo 转移灶大小 免疫组化 whether pharmacologic inhibition of the PDGFRb pathway recapitulates the effects of PDGFRb or mutant p53 depletion? 时效关系实验 量效关系实验 Crenolanib两种细胞IC50 whether crenolanib can suppress metastasis? Crenolanib处理与DMSO处理 荧光和免疫组化 细胞凋亡 KPflC 细胞 pancreatic cancer cells can provide a source of PDGF ligand that could trigger autocrine activation of PDGFRb 条件培养 Summary 5 abrogation of autocrine activation signaling of PDGFRb leads to a significant reduction of invasion and metastasis driven by mutant p53 . whether PDGFRb inhibition prevents the development of metastasis in KPC mice? Question * Journal: Cell Reporter: XuWang IF: 33.116 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 Ideas Mutant p53 maintain metastatic phenotype the relationship between Mutant p53 and PDGFRb on metastasis PDGFRb madiates metastasis when Mutant p53 was depleted imatinib can inhibit metastasis by targeting PDGFRb PDGFRb correlates with disease-free survival PDGFRb as a Downstream Mediator of Mutantp53 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 w