ACTA LASER BIOLOGY SINICA ,P53 P53. Studying the Application of P53 Gene in Tumor Therapy [1 ] 2 P53

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1 ACTA LASER BIOLOGY SINICA Vol. 10 No. 1 Mar P53, ( ) :,P53,P53,, P53, P53 :P53 ; ; ; :R :A : (2001) Studying the Application of P53 Gene in Tumor Therapy XU Chao - yang, TAN Shi - ci, XING Da (Laser life science institute South china normal university Guangzhou Guangdong,China) Abstract :More and more research indicate that P53 is a sequence2specific transcription factor,whose transcrip2 tonal targets induce growth arrest and apoptosis. So most scholars agree that P53 is a tumor suppressor gene. This paper mainly introduce the structure,function and action mechamism of the P53 gene. Key words :P53 ;tumor therapy ; GFP ;LSCM P53,,P53 : P53 N (1,, 43 ),, P53 ( mdm2, TAF40, TAF60) P53, P53, ;P53 DNA, DNA ; 1 P53 P53 17 (17P13), 2 P53 2 P53 53KD 393 [2 ( ] P53 P53) P53 11, 5 P53, P , 8 1, P53 2 DNA, DNA 2 SV40 large tumor antigen P53 P53 (nuclear localization signal) P21wafiΠcip l G 1, (oligomerization domain) P53 (phosphorylation site) P53 C P53 DNA, DNA DNA [1 ],P53 :

2 Fig. 1 : Structural features of the P53 tumor suppressor gene. The transcriptionactivation site ( TAS),heat shock protein binding site ( HSP),SV40 large T2antigen binding sites(sv40), adenovirus E1b and papillomavirus E6 binding sites,cellular Mdm2 binding site,nuclear localization signal(nls),oligmerization domain (OLIGO) and phospho2 rylation sites(cdc2 and CDK). The five evolutionarily conserved domains are labeled HCD I2V and the hot spot regions are HSR A2D. 1 : P53 (TAS) (HSP) SV40 T2 (SV40) E1b E6 Mdm2 (NLS) (OLIGO) 5 HCD I V, HSR A D [3 ], 200,, 50 % P53, P53 [4,13 ],P53 P53, 4 ( 1,A D), P53 DNA, Gadd45 P53 19KD [9 ],P53 Gadd45 [5 ], P53 P21 Cip 1, P53,, P53, P21 [6 ], (Makri D. P53 ) P53 bcl22 P53, Π :, Bcl22, [10 ] Bax, [10 ], P53 Martin Bennett, P53 P53 P53 P53 P53 DNA,,, (death receptor),p53,p53 P53, P53 P53 FasL, [7 ], P53,, P53 P53 :P53,, P53

3 1 :P53 73 CD95, P53, P53 CD95 FADD, CD95, P53 (cytoplas2, P53 CD95 mic sequestration) P53 [14 ],P53 G 1, 37 % P53 (, G 2 ΠM P53 ), P53 P53 G 2 ; P53,, P53 DNA, (neuroblastoma) P53 G 2 ΠM, 3 GFP, GFP MCF27 RKO LSCM P53 Knippschild U [11 ] LSCM, P53 ) : GFP2CKI delta GFP2CKI epsilon ( delta and epsilon isoforms of casein kinase I CKI2delta CKI2epsi2 lon,cki delta CKI epsilon GFP GFP2,Lys2305 Ala P53 CKI delta GFP2CKI epsilon) ( cytoplasmic sequestration), Lys2305 P53 N CKI delta CKI epsi2 lon P53NK ( P532N2terminus2tergeted protein kinase) (Fig2B. e) Lys2305 P53 P53 P53 P53 CKI delta CKI epsilon Shun2Hsin Liang [1 ] He P,Tang Z [12 ] PEGFP2N1 PEGFP2P53 MHCC97 ( ) LSCM, PEGFP2 P53K305N ( C P53 Lys2305 P53 ), N1 MHCC97,, PEGFP2P53 MHCC97, P53K305N P53 ( Fig2B. h) P P53, P21ΠCIP1ΠWAF1 mdm2 GAAD45 cyc2 lin Bax IGF2BP3, P53 Shun2Hsin Liang [1 ] P53 P532 ( P53,P53 ( Fig2B. a,b) ; P53 Ala Asn Glu Thr, P53 P53NK NLS Lys2305 P53 P53 (P53K305N) C ( Fig2A) P53K305N ( Fig2B. f g), Lys2305 P53 (cyto2 plasmic sequestration) Fig. 2 A., The C2terminal amino acid sequences of the human P53 protein. The nuclear localization signals(nls) are underlined. 2A, P53 C2 (NLS) Lys2305 P53, CSD (cytoplasmic sequestration domain, P53C 326

4 signals. P53 C ) NLSI, P53 (importin) P53 NLSI,NLSI importin Lys2305 importin NLSI,Lys2305 P53,Lys2305 CSD, Lys2 305,CSD NLSI 4 P53,P53 ( Fig. 2B. Alternations in the subcellular localization, ) of mutated P53 proteins. Fluorescent pictures showing, P53 different subcellular localization between wild2type and mutated P53ΠGFP fusion proteins using LSCM. Distribution of wild2type P53ΠGFP in MCF27 and RKO cells is,, P53 P53,P53,P53 shown in ( a) and ( b), respectively. MCF27 cells were transfected with the fusions including P (c), P (d), P53K305N References ( e ), P53K305N ( f ), P53K305N ( g ) and [1 ] Shun - Hsin Liang,David Hong,and Michael F,et al. Coopera2 P53K305N (h). The fluorescence was examined at 48 h after tion of a Single Lysine Mutation and a C2treminal Domain in the transfection Cytoplasmic Sequestration of the P53 Protein[J ]. J Biol Chem, 2 B. P , 273(31) : LSCM, P532 [2 ] Kastan M. B,Onyekwere O, Sidransky D et al. Participation of GFP a b P53 protein in the cellular response to DNA damage[j ]. Cancer P53ΠGFP MCF27 RKO P53 Res,1991,(51) : (P ( c), P ( d), P53K305N ( e), P53K305N [3 ]Oren M. P53,the ultimate tumor suppresser gene [J ]. FASEB J. (f),p53k305n (g) P53K305N (h) GFP 1992 (6) MCF27 [4 ]Lane D P. P53,guardian of the genome[j ]. Nature,1992 ;358 : ) Lys2305, [5 ]Liu T J, E1 Nagger A K,J,et al. Apoptosis induction mediated CSD Lys2305 P53 5 by wild type P53 abenoviral gene transfer in squamous cell car2, ( cinoma of the head and neck [J ]. Cancer Res,1995,55 :3117. Xenopus Lys2305 ), P53 [6 ]Li G,Manning A M,Hanlon KA, et al. Sodium butyrrte induces NLS ( nuclear localization apoptosis in human colonic tumor cell lines in a P53 indepen2 P53, P53 P53 P53 [1 ], P53,,,,, P53 P53 [5 ] P53 P53, P53 [1 ],P53

5 1 :P53 75 dent pathway implications for the possile role of dietaty fibre in the prevention of large bowel cancer [J ]. Int. J. Cancer,1993, 55 :498. [7 ] Midgley C A, Owens B, et al. Coupling between gamma irrdiation, P53 induction and the apoptotic response depends upon cell type in vivo[j ].J. of Cell Science,1995,108 :1843. [8 ]Makri D,Schuly W A,et al. WAF1Πp21 regulates proliferation, but does not mediate p532dependent apoptosis in urothelial car2 cinoma cell lines. [J ]. Int.J. Oncol. 1998,12(3) :621. [9 ]Smith M L,Chen I T,Zhan Q M,et al. Interaction of the p53 up2 regulate protein Gadd45 with proliferating cell nuclear antigen [J ]. Science,1994,266 :1376. [ 10 ]Martin Bennett, Kirsty Macdonald,Shiu - Wan Chan,et al. Cell surface trafficking of Fas : a rapid mechanism of p532mediated apoptosis Science,1998,282(5387) :290. [11 ] Knippschild U,Milne DM,Campbell LE,et al. P53 is phospho2 rylated in vitro and in vivo by the delta and epsilon isoforms of casein kinase 1 and enhances the level of casein kinase 1 delta in response to topoisomerase2directed drugs[j ]. Oncogene 1997 Oct 2 ;15(14) : [12 ]He P,Tang Z,Ye S,Liu B. The expression and localization of wild2type p532gfp fused gene on human high2metastasis hepa2 tocellular carcinoma cell line [J ]. Chung Hua Kan Tsang Ping Tsa Chih 2000 Apr 20 ;8(2) : [13 ]Paxel G. Komarov,Elena A. Komarova,et al. A chemical inhib2 itor of p53 that protects mice from the side effects of cancer therapy[j ]. Science, ;285(5434) : [14 ]F. Bunz,A. Dutriaux,C. Lengauer,et al. Requirement for p53 and p21 to Sustain G 2 arrest after DNA damage [J ]. Science. Volume 282,pp :, ; , Biography XU Chao - yang :male,born in 1976,got the Bachelor degree from South China Normal University in Now he is studying for Master degree in institute of laser life science of the same university. Tutor is professor TAN Shi - ci and professor XING Da. E2mail :XUCY@ scnu. edu. cn