外源p53基因和抗血管生成小肽F_省略_乳腺癌移植瘤生长和转移的抑制作用_肖绍文

论著外源p53基因和抗血管生成小肽F56联合对小鼠乳腺癌移植瘤生长和转移的抑制作用肖绍文1,颜　博1,杨敬贤1,寿成超2,张珊文1△(　1.,2.,　100036)[　]目的:p53F56。方法:BICR-H1NOD/SCID。,Adp53+F56、Adp53、F56,NOD/SCIDAdp53+F56、Adp53、F56、Adlacz,p53、F56,,NOD/SCID(Adlacz)。、,p53、(vascularendothelialgrowthfactor,VEGF)(microrasculardensity,MVD);NOD/SCID。结果:p53、F56,,Adp53+F56、Adp53、F562.47、4.37、4.6912.49;,Adp53+F56Adp53p53,P539.4%、6.3%,,(P=0.693);Adp53+F56、Adp53、F56VEGF,VEGF21.9%、9.4%3.1%,,(P=0.284);Adp53+F56、Adp53、F56MVD14.50±2.54、16.28±3.4418.06±7.66,(24.93±6.53),(P=0.000),Adp53+F56、Adp53、F56,Adp53+F56。NOD/SCIDAdp53+F56、Adp53、F561.143±0.378,2.750±0.886,3.375±0.518,Adlacz(5.000±0.816)(5.670±0.817)(P=0.000),Adp53+F56。结论:p53F56。p53p53VEGF。[],p53;;;[]R737.9　　[]A　　[]1671-167X(2007)02-0171-06InhibitoryeffectofAdp53andF56ongrowthandmetastasisoftransplantationofthebreasttumorinmiceXIAOShao-wen1,YANBo1,YANGJing-xian1,SHOUCheng-chao2,ZHANGShan-wen1△(1.DepartmentofRadioationOncology,2.DepartmentofBiochemistry,PekingUniversitySchoolofOncology,Beijing100036,China)ABSTRACT　Objective:ToinvestigatetheeffectsofAdp53andF56onthegrowthandlungmetastasisofbreastcancer.Methods:TheBICR-H1cellswereinoculatedintothemammaryfattypadofBALB/CnudemiceandNOD/SCIDmicetoestablishbreastcancermodel.ThenthenudemicewithxenografttumorwererandomizedintogroupAdp53+F56,Adp53,F56andcontro.lTheNOD/SCIDmicewithxenografttumorwererandomizedintogroupAdp53+F56,Adp53,F56,Adlaczandcontro.lTheyweretheatedfor3weeksaccordingtotheplan,diversityofthevolumeandhistopathologyofxenografttumorofnudemicewasobservedandtheexpressionsofp53andVEGFgene,andmicrovesseldensity(MVD)weredetectedbyimmunohistochemistry.LungmetastasisofbreastcancerinNOD/SCIDmicewasob-served.Results:(1)IntratumoralinjectionsofAdp53,F56,andtheircombinationresultedinaninhi-bitiononthegrowthofxenografttumorofBICR-H1cells.TheultimaterelativegrowthvolumesofgroupsAdp53+F56,Adp53,F56andcontrolwere2.47,4.37,4.69and12.49respectively.(2)Aftertreat-ment,P53positiverateofgroupAdp53+F56,Adp53increased9.4%,6.3%thanbeforerespectively,:(2002AA216111)SupportedbyTheNationalHighTechnologyResearchandDevelopmentProgramofChina(2002AA216111)△Correspondingauthor’se-mail,[e-mail]171()JOURNALOFPEKINGUNIVERSITY(HEALTHSCIENCES)　Vo.l39　No.2　Apr.2007butcomparedwithcontrolgroup,thedifferenceisnotsignificant(P=0.693);VEGFproteinofgroupAdp53+F56,Adp53andF56decreased21.9%,9.4%and3.1%thanbeforerespectively,butcom-paredwithcontrolgroup,thedifferencewasnotsignificant(P=0.284).NecrosisanddecreaseofvesselinthetumorandmorphologicalchangeofendotheliumwereobservedunderlightmicroscopeinthegroupsAdp53+F56,Adp53andF56.MVDestimatedbyFⅧ-RAstainingofgroupAdp53+F56,Adp53andF56were14.50±2.54,16.28±3.44and18.06±7.66,comparedwithcontrolgroup(24.93±6.53),thedifferenceissignificant(P=0.000).(3)TheaveragenumberoflungmetastasisofNOD/SCIDmiceingroupAdp53+F56,Adp53andF56were1.143±0.378,2.750±0.886and3.375±0.518respectively,lowerthanAdlaczgroup(5.000±0.816)andcontrolgroup(5.670±0.817)obvi-ously(P=0.000).Conclusion:Adp53combinedwithF56cangreatlyinhibitgrowthandmatastasisofbreastcancerinvivo.Themechanismofanti-tumoreffectsofAdp53andF56mayberelatedtotheanti-angiogenesiseffectonmalignanttumorthroughinhibitingtheexpressionandactivityofVEGF.KEYWORDS　Genes,p53;Vascularendotheliagrowthfactors;Breastneoplasms;Adenoviridae　　、。,p53,[1]。,(vascularendotheliagrowthfactors,VEGF),,VEGF[2-4]。p53(Adp53)E15,p53(CMV)、p53(cDNA)SV40,(GFP)。Adp53VEGFF56,。1　1.1　BICR-H1。10%()RPMI1640。,P53。,p531048G-A。293,E1,,10%DMEM,。1.2　BALB/Cnu/nu,,4～5。NOD/SCID,,4～5。,。(specialpathogenfree,SPF)。1.3　、1.3.1　Adp53　。(Adlacz)。Adlaczp53,Adp53,SV40β-gal。1.3.2　、[5,6]　Ad-p533.0×1012pfu/mL,Adlacz1.0×1012pfu/mL。1.4　F56VEGFFlt1(VEGFR1),,90%。50μL(30μg)[7]。1.5　p53Do-1(SantaCruz),VEGF(SantaCruz)F-Ⅷ(Zymed),。1.6　Ad-p53F56BICR-H15×106(0.2mL)BALB/C,Adp53+F56、Adp53、F56,4。8～10mm,。,Adp5350μL(1×1010pfu);F5650μL(30μg),50μL。Adp5324hF56,36。,1,π/6[+)/2]3。,(,1),。2。1.7　P53、VEGF(mi-crovasculardensity,MVD)1.7.1　BICR-H1p53VEGF　BICR-H110%RPMI1640172()JOURNALOFPEKINGUNIVERSITY(HEALTHSCIENCES)　Vo.l39　No.2　Apr.20071∶1(),,,37℃5%()CO224h,。,SPp53VEGF。1.7.2　P53、VEGFMVD　48h,,,,P53;4%(),HE,(SP)VEGFMVD。。1.7.3　　P53。5400200。Fromowitz():0;1;2;3。:5%0;5%～25%1;26%～50%2;51%～75%3;75%4。:2(-);2～3(+);4～5(++);6～7(+++)。(++)(+++)。VEGFP53。Weidner[8]。40(“”),200,1。5,MVD。1.8　Adp53F56NOD/SCID4～5NOD/SCID40,BICR-H1,0.2mL(5×106)。2Adp53+F56、Adp53、F56、Adlacz。10,300mm3。Adp5350μL(1×1010pfu),F5650μL(30μg),Adlacz50μL(1×1010pfu)。Adp5324hF56,50μL。36。56d,。,4%,,4%,HE。1.9　SPSS11.5。MVD±(–x±s),;P53VEGF。2　2.1　Adp53+F56BICR-H12.1.1　BICR-H1　。10d,96.9%(31/32)。48h,(1)。,Adp53+F56、Adp53、F562.47,4.37,4.6912.49。2.1.2　Adp53+F56BICR-H1　,,Adp53+F56。Adp53+F56F56Adp53(2,3)。1　BICR-H1Figure1　RelativegrowthcurveofxenografttumorofnudemicewithBICR-H1cells2.2　P53、VEGFMVDBICR-H1P53,VEGF(4)。p53,,Adp53,P53(1,5),,。F56p53VEGF,。F56P53MVD,。Adp53+F56F56(6)。2.3　Adp53F56NOD/SCID,,4NOD/SCID(2Adp53+F56、Adlacz1)。Adp53+F56、Adp53、F56173,　p53F56,;Adlacz,,1。,,,。1,Adlacz1,。NOD/SCID2。A,control;B,Adp53;C,F56;D,Adp53+F56.2　Figure2　Dissectionpicturesofxenografttumorofnudemice3