·498"华西口腔医学杂志第32卷第5期2014年10月WestChinaJournalofStomatologyVo1.32No.5Oct.2014http://www.hxkqyxzz.net【文章编号J1000—1182(2014)05-0498—06近红外量子点连接的精氨酸.甘氨酸.天冬氨酸肽段荧光探针对口腔鳞状细胞癌的活体可视化成像白云龙黄昊杨凯唐洪重庆医科大学附属第一医院口腔颌面外科,重庆400016【摘要l目的探索用精氨酸.甘氨酸.天冬氨酸(RGD)肽段连接的近红外量子点荧光探针对口腔鳞状细胞癌(OSCC)的原位可视化成像情况。方法将含有RGD序列的肽段与发射波长为800nm的近红外量子点(QD800)偶联,制备QD800.RGD荧光探针。将人颊鳞状细胞癌BcaCD885细胞植入裸鼠颊部皮下建立OSCC模型。用QD800-RGD探针和CD105单克隆抗体对OSCC冰冻切片行直接免疫荧光双重染色,用激光扫描共聚焦显微镜观察QD800.RGD探针与肿瘤新生血管内皮细胞表达的整合素0【,,p的结合情况。将QD800.RGD探针通过尾静脉注射JLOSCC模型动物,在不同时间点通过活体成像观察QD800.RGD对OSCC活体原位可视化动态成像情况。在12h后处死荷瘤鼠取出肿瘤,检~JQD800.RGD在体内与肿瘤新生血管内皮细胞表达的整合素p的结合情况。结果QD800一RGD探针在体内和体外均能与oscc]~瘤新生血管内皮细胞表达的整合素,p,特异性靶向结合。静脉注射QD800.RGD探针后能对体内OSCC进行清楚地可视化成像,在注射QD800.RGD后0.5~6h内肿瘤成像最完整,信噪比最高,9h时肿瘤荧光强度显著减低,但在12h时仍能看到明显的肿瘤成像。结论以肿瘤新生血管内皮细胞表达的整合素%13为靶点,利用QD800.RGD探针经静脉注射后能对OSCC进行清晰地可视化成像,在OSCC的诊断和个体化治疗等方面有巨大的发展前景。I关键词】口腔鳞状细胞癌;量子点;整合素0【p;精氨酸.甘氨酸.天冬氨酸肽段;成像I中图分类号】R739.8【文献标志码lA【doil10.7518/hxkq.2014.05.017Insituvisualimagingoforalsquamouscellcarcinomainmicebyusingnear-infraredquantumdotsconjugatedwitharginine-glycine—asparticacidpepfidefluorescentprobesBaiYunlong,HuangHao,,辔Kai,TangHong.(Dept.ofOralandMaxillofacialSurgery,ThefAffiliatedHospitalofChongqingMedicalUniversity,Chongqing400016,China)【Abstract】ObjectiveToinvestigateinsituvisualizationusingnear-infraredquantumdots(QDs)conjugatedwitharginine—glycine-asparticacid(RGD)peptidefluorescentprobesinoralsquamouscellcarcinoma(oscc).MethodsQDswithemis—sionwavelengthof800nln(QDS00)wereconjugatedwithRGDpeptidestoproduceQD800-RGDfluorescentprobes.HumanOSCCcell1ineBcaCD885wasinoculatedinnudemicecheekstoestablishOSCCmousemodels.FrozenBcaCD885tumorsliceswereimmunofluorescencedoublestainedbyusingQD800·RGDandCD105monoclonalantibodyandwereobservedusingalaserscanningconfocalmicroscope.QD800-RGDwasi~ectedintotheOSCCmodelsthroughthetailveins,andtheinsituvisualizationwasanalyzedatdifferenttimepoints.Themiceweresacrificed12hafterinjectiontoisolatetumorsfortheexvivoanalysisofprobelocalizationinthetumors.ResultsQD800-RGDspecificallytargetedtheintegrinav133expressedintheendothelialcellsoftumorangiogenicvesselsinvitroandinvivo,producingcleartumorfluorescenceimagesafterintravenousinjection.Themostcompletetumorimageswithmaximalsignal-to—noiseratioswereobserved0.5hto6hafterinjectionoftheprobeandsignificantlyreduced9haftertheinjection.However,thetumorimagewasstillclearlyvisibleat12h.ConclusionUsingintravenouslyinjectedQD800-RGDgenerateshighqualityOSCCimageswhenintegrinav133,whichisexpressedintheendothelialcellsoftumorangiogenicvessels,isusedasthetarget.Thetechniqueoffersgreatpotentialin【收稿日期【基金项目【作者简介【通讯作者2013.12-25;l修回日期】2014-04—21国家自然科学基金资助项目(81172205)白云龙,硕士,E.mail...