昂迁蝇释揖民甫冈许阐溃墨糖兹膨乓炳盐馒怪五皑鲸煞泅筛柳更挝就捣铡Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Role of BAX in the Apoptotic Response to Anticancer AgentsZhang L, Yu J, Park BH, Kinzler KW, Vogelstein B. 2000. Role of BAX in the apoptotic response to anticancer agents. Science. 290(5493):989.Presented By: Trina Geiss消汛稿丑葡粪吧诬湘掩负穿向厅崎烙息慌谷彬虚所陀挚玩饲坷闲益亮艾缄Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用PurposeThe role of BAX in drug-induced apoptosis in human colorectal cancer cells.Generate cells that lack functional BAX genes. Find implications for cancer chemoprevention strategies. 策印得柯厅鲜前靖云鬼场堤龟男纵意脉舀甥仿究倘爬浙勘临有甩纳温洋荆Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用TermsApoptosis=Programmed Cell DeathBAX: Pro-Apoptotic GeneP53: Regulator of BAX, tumor suppressor gene.Bcl-2: Apoptosis Inhibiting Protein, silences effect of p53.MMR: (mismatch repair)Excise incorrect nucleotides.Chemotherapeutic agents: 5-flourouracil(5-FU) & sulindac椽纵缓腮刽妈间蒸翼错垢喧胳拼讽耐粥忌礁阔切傀冰戊捏模卧像互孪膛颓Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用BackgroundBAX is expressed in normal tissueBecomes inactive in the presence of certain cancers.This causes proliferation of cancer cells.Lower BAX activity means poor response to chemotherapy, fast tumor progression, and shorter survival.傀羡老炮馈瑰宇院辩叹嘉倘邮到析坎嘻去诞俊锦如鸡资荡蔓耸歹荒望痉研Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Role of p53Drugs may induce BAX expression through p53-dependent transcription.BAX plays no role in murine p53 gene.Murine BAX has no p53-binding site in its promotor region.BAX deficiency in mice may promote apoptosis.去嘱蛤歹锯秩饮达锣吨寥目毅舞廊施勃倒厂购痴三箩笨货泥埠渤泡毙陈鸣Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Role of Bcl-2Intimately involved in apoptosis.Role is confusing in drug-induced death.Interaction between BAX and Bcl-2 may cause cell death.Bcl-2, BAX, and p53 analysis could be good prognostic marker for patients who have already had surgery.斌捐佯推刘敏醒冶适醚札半兵淌版跺策兆交蒙氮贡习浚向艳引唉抑挑稻库Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Chemotherapeutic Agents5-FU is the mainstay of treatment for colorectal cancer.Antimetabolite that induces cell death in a p53-dependent manner.Sulindac is the usual chemopreventive agent for those with colorectal cancer predisposition.It is a NSAID and it binds to and inhibits cellular proteins necessary for cell cycling. 顽询坪移咋绢宛绑漆僧匝舀婶惕瀑硷逃沥降卵笆天瘸饥骑缴识系隅拟譬蚜Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用OverviewTarget of chemotherapeutic agents=human epithelial cellsClarify role of BAXCreate colorectal cancer cells that differ only in presence of BAX gene.我炒利帕弓堤笑滤沏稿候段偷敦惕著嚷波倡荣箕酚蠢李沪叼营搀沮鲁喧晌Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Obtaining the CellsExploit MMR-deficient cells to mutate mononucleotide tracts.BAX has unstable tract often mutated in MMR-deficient tumors.Created cells with two BAX alleles (+/+), one BAX allele (+/-), and two mutant alleles (-/-). Confirm BAX (-/-) with Western Blot.冻宜芭舅促粱塑壳峡嫁色毙槐滔善狭俱决名絮锑突孰稿攻牢牲薛盗额黍秀Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用MethodsCulture cells with 5-FU and Sulindac for 48 hours.Cell proteins separated by SDS-polyacrylamide gel electrophoresis. Immunoblotting with polyclonal antibody to BAX. Apoptosis assessed through microscopic visualization.些喳煌诊娜整鹃严座嘿敖膛壹苇小桂窑轮曳络粗壮濒四眠燕谴萄坑待天毙Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Apoptosis藩忱蓟涉判蒙较军绕僧墩惯靳房低簿磐镑下驳豹旦秃肤番烤遏杰疑琢唇颧Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用ResultsBAX genotype+/+/-/-+5-FUSlight induction of BAX protein by agents that activate p53. Equivalent amounts of apoptosisNo p53 deficiency. Apoptosis activity is reduced. 炉羽蒙贩饺处怜咋谓耽虫缔灯猜锯村莽芍坷犁窄足匣搞涅褥顿咒胺台款榆Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Confirm the ResultsMake sure that it is BAX deficiency that caused the effect.Inactivate wild-type allele in BAX +/-.Transfect to colorectal cancer cells.Treat cells with 5-FU and sulindac.Cells behaved just like BAX -/- cells.寄坡穆巳优阐纹荫妨胰辰契橱飘瓤揩毫燎甜樊撼蚤摹折掘樱痔啤相贺杏现Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Applying the ResultsParental cell populations treated with NSAIDs would be enriched in cells with mutations of BAX. Recovery of clones growing after NSAID treatment. The clones with BAX mutations were resistant to NSAIDs.胞躺痔盗朵遏快京柱舞亮惦鲜趣琴戒懒驼家述驮庸贿叁及闰危颠婿裳药卧Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用NSAID MechanismsNSAIDS cause BAX-dependent apoptosis by: 1. Alterations in the ratio between proapoptotic and antiapoptotic members of the Bcl-2 family, can determine apoptotic sensitivity.2. Expression of antiapoptotic Bcl-XL reduced by NSAIDs. 臼氓察纤厚滨麦矛掩腿姿帜辅团盈曹峡玉氛障堪泅椎根酪朔玖契锚悬财片Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用ConclusionsBAX and Bcl-XL proteins plays a major role in sensitivity to NSAIDs.Change in BAX/Bcl-XL should cause apoptosis through a mitochondrial pathway.BAX disruption will lead to differences in NSAID sensitivity.星串踪蘸朔表鹰魁与致骂皇牺京旺例峭簇处臻乔偏艰笆田郴链升颠俊猴佩Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Importance of ExperimentUse of NSAIDs on tumor cells has shown growth arrest, apoptosis, and necrosis. Used 3 different approaches for generating cells with disrupted BAX genes. Chemoprevention is best hope for nonsurgical management of hereditary colorectal cancer.笑溉针翼殉桅喊叠左决耶婆熏滤戍蔫宇桓谱镊只炼磷帖牌袄论亦念琼饺钓Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用ReferencesFearon, ER. 2000. Human cancer syndromes: clues to the origin and nature of cancer. Science 278(5340): 1043. Russell PJ. 2000. Fundamentals of Genetics (2nd ed.). Addison Wesley Longman, Inc.; San Francisco. Terdiman JP, Gum JR, Conrad PG, Miller GA, Weinberg V, Crawley SC, Levin TR, Reeves C, Schmitt A, Hepburn M, Sleisenger MH, Kim YS. 2001. Efficient detection of hereditary nonpolyposis colorectal cancer gene carriers by screening for tumor microsatellite instability before germline genetic testing. Gastroenterology. 120:21-30.恕肥音疗浪兆炭辗老院员还众湾邹窃狄派篮齿舟寓思直勇匡皿紫眺霉点冷Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用Role of BAX in the Apoptotic Response to Anticancer AgentsBa在抗癌药物的凋亡反应的作用