肿瘤凋亡 Apoptosis and cancer课件.ppt

I decided to look for biochemical alterations that would distinguish normal T cells and preapoptotic T lymphocytes. pH, cation concentrations, membrane permeability, lysosome change and so on. DiOC6, a dye that indirectly measures the mitochondrial transmembrane potential(JEM,1995) Loss of MTP marked a irreversible early step of the apoptotic process(JEM,1995) A cell free system, in which we confronted purified mitochondrial with purifed nuclei. Upon induction of permeability transbility transition, we found that mitochondria released an apoptosis inducing factor that cause chromatin condensation and fragmentation when added to purified nulclei.(JEM,1996) The mitochondrial release of these apoptogenic factors but not their action on the nucleus was found to be controlled by Bcl-2.(JEM,1996) Mitochondrial would decide the fate of cells and serves as a central integrator and coordinator of the apoptotic process.(FASEB J,1995) Mitochondrial are bull shit !? Without this help, we would have been forced to abandon our work Xiaodong Wang Douglas Green Cell free system of xiaodong wang Cell free system of kroemer To detect caspase activation as the final readout To look for a protein that would cause chromatin condensation, in the absence of cytosol, in a caspase-independent manner. AIF (Nature,1999) Artefact-inducing factor? It is probably wrong to concentrate on just one or few proteins (ANT, VDAC,p53) as being crucial regulators, we should, at least sometimes, concentrated on processes, rather than on individual molecules. Cyto C, How apoptosis is regulated Tsujimoto和Crose发现极大多数非何杰金氏滤泡状B细胞淋巴瘤(B-cell lymphomas)存在染色体易位t(14;18)。 caspases Apaf-1 ? 14号染色体的IgH基因—Bcl (B cell lymphoma) -2基因 基因转染实验显示Bcl-2高表达广泛抑制大多数刺激剂诱导的细胞凋亡。 Bcl2 含有1-4个Bcl-2同源结构域（BH1-4）。分两类：抗凋亡基因，如：Bcl-2；促凋亡基因，如：Bax。 促凋亡蛋白能引起线粒体外膜通透性改变，导致cyt c 释放，引起凋亡。 bcl-2相当于线虫的ced9。主要定位于线粒体膜、也存在于ER和外核膜，拮抗促凋亡蛋白的功能。 bcl-2基因家族 Bcl-2 gene family（Zimmermann 2001） Early work on the function of Bcl-2 David Vaux Full leng