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1、肿瘤研究中常用的方法11分子生物学细胞生物学UL4组织病理学动物模型分子生物学RNARNA的提取RT-PCR/PCRReal-Time-PCRNorthern blotPROTEINWestern blotImmunology?细胞生物学一肿瘤细胞增殖HJ中瘤细胞凋亡-A细胞计数、MTT/结晶紫染色-流式细胞仪检测周期AnnexinV/PI 双染Trans we II肿瘤细胞迁移T小鼠肿瘤组织切片（石蜡切片、冰冻切片）人体肿瘤与癌旁组织切片H&E （苏木精伊红）染色免疫组织化学免疫荧光（Confocal）H&E （苏木精伊红）染色免疫组织化学免疫荧光（Confocal）e裸鼠成瘤实验m药物诱导
2、成瘤实验#THE JOURMAl Or SOLOGXZAL CMEV VCL 237 M2 .21, =2 r 7737-177-15. Vay U. 20122二yTxdTTir二工三彳二丁三工七一二了白31三二丁三eogy 二 pjo 厂曰d n rc J 5 AForkhead Transcription Factor FOXO3a Protein ActivatesNuclear Factor kB through B-cell lymphoma/leukemia 10 (BCL10) Protein and Promotes Tumor Cell Survival in SerumD
3、eprivation*Re*”配忸publit式匕也削如立旧,2311 in 值屈忆 fcim,. Warch 31,. 2012 Fiblisbed.JBC Papersir Press, April2.1C12. COI10.1074/bc,M 111.2917C8#Non-canonical stimuliCanonical stimuliTRAF3“4 ”s- TRAFsTRAFs& othersCanonical NF-kB Pathwaycanonical 同 hweiynoncanonicEil 口 sitlwsiyTNF-a, IH fLPSPCD4OLLT-OChPnLPS,
4、 CD40L.BAFRRAKT PATHWAY0coAKTPDK2PTEN :SurvivalTransformationRAC1/0DCGrowthTranslationApoptosisCytoskeletal rearrangementTransformation Ceil cycteCelbcycte airest Glucose metabolsm ApoptosisDNA repairNature Reviews | Cancer10FOKCgMV常 tin2g3T 工备 PQ3Relative Lugi activity (Luci/p-gal)DORjelative Luci
5、activity(LucS/p-gal)oRelative TN Fee mRNA LevelRelative Luci activity(Luci/p-igal) CQms 一 SIRNELIBSsNA2 (X15Q13 (X1328T HCT1 二 6 AggGon siRNA ”F0XO30 FQXQSaTmB-n Fo*03a B&S51 3x0331 0七G 二 nF-GURE 1 Fox。3a act-vaars NFKB293T 工CTH A549CCyto NuclsiF0X03a-1 _ + _ _ + _s i F 0 JK 03 a-j? , , + 十#DFOXO3aT
6、M - 十i& L 一!F0X03aTM 一| pactin 1TsiF0XO3a 一 十FCX03a |一 1lactin匚mpty ,吟纨M FOXOS0TMControl kBaSR(而 E 母nip2?B母vrrl 学等ffio:EIPJKKp InputConiroi F0x03aTM号尹wffiHControl IKKp-DNFIGURE 2. FOXO3a Induced NF-kB Activation through IKKxi时(吊旨n旦 目芝看用pn_JBa苗怔ai& d 2 0 者=2w一灯 HE Ils(而菖乡旦*BsaB 口n，vAnE a工*FOXOSaTMBCL
7、10GAPDHBCL10InsulinC 432CAGCCTGGGCGACAGAGTGAGATCCTGTCTCA-385TAAA TAAA TAAA 7AATC 昌 GTTG ARTGCATACTGTGTGCCCAACTACCTTTAAGAACTTTACATATGATCCCGG CAGCCCTAAAAGG TAAGC A TATCTATCTCCATTACACAGATTAACTAATGAGGAAAAGCTAT-255GTraTAATGAACAGTGACCAAAAAGTATGCAAACGTTAAATGAGGTGTTAAAGTGGTGGGATTGTAGGTGGECTTTTTTTCECAA/MWTrT
8、GTTTAGT -141ATTGTAGTTAGGCTGTTTGTGTGTATGCATGTGTATGTGTGTATGAGTGCGTTTGTGTGTGACCAAAACAGTAACTGGCAAACGTAAGAAGAGGAAATTGTAgggcagggtctgggaaaaggggaggaacccaa4dGGTACTGACAAGCCCAGACCGGACTCCCsiFOXO3a-1siFOXO3a-2BQL1029订FOXOBasiFOXQSad siFOXO3a-2BCL10F0X03a 卜3d的HCT116(3-actiniBCLIWacti n 1 0.46 0 35cmseJ) 占ts加 h2n1
9、v&:-EHBCLIO/actin 1 0_B7 0.35(pot立者p& 力NLLN也皂：IE币工FIGURE 3. FOXO3a activates NF-kB throughCLIORelative Luci activity A (LodJip-gal)一 432;1口 coni MyqFCJXOHalRelative Ludi ActivityCFirefly/Rinflte)BCL10 Prom2rER)苧 comd一 FOXO3ZTM+.JLII255J+-1Relative Lua admlyLuci/pal)5鹤匚手Nuc-mmFactWT prowMHaapabmsh三十
10、II +F-GURE 4, Fox。3a regu-ates expression of BCL10 -ranscriptiona=y , 14HCT116Oiti TNFtrp-FOXO3aFCXO3BWinCon IL-1|1293 TSErurripF0X03aFOXOSsp-acLin匕一居3C 33J-5考Ex运fc竺予一百 一：irq ?7ir5”urn (+)#293THCT116亘百祀不言=巴 2言一号？君之Conlrol sirFOXO3s前三正圆更fe营杳 nn-lluA一靠-luHFIGURE 5. FOXO3a activates NF-kB under serum-free conditions
