%0 Journal Article
%T 用噬菌体展示技术筛选喉癌细胞靶向肽的研究
%A 冯俊
%A 李丽
%A 杨洪斌
%A 刘世喜
%J 华西医学
%P 1844-1847
%D 2011
%X 【】　目的　筛选人源喉癌Hep-2细胞株特异结合的短肽，作为喉癌靶向治疗的载体。　方法　体外培养Hep-2细胞株作为靶细胞，人正常喉黏膜上皮细胞为吸附细胞；用噬菌体展示十二肽库进行3轮差减筛选，随机挑取10个噬菌体克隆进行测序；采用酶联免疫吸附（enzymelinkedimmunosorbentassay，ELISA）法鉴定噬菌体与Hep-2细胞的结合活性；通过免疫荧光鉴定喉癌细胞特异性结合肽（F2）噬菌体阳性克隆与喉癌细胞结合的特异性。　结果　经过3轮筛选后，噬菌体在靶细胞Hep-2上出现明显富集；ELISA分析鉴定显示5个阳性克隆能与Hep-2细胞特异结合，其中F2噬菌体克隆对喉癌细胞的结合靶向性明显高于对照细胞（P<0.05）；免疫荧光显色显示，F2能特异性地与喉癌细胞结合。　结论　利用噬菌体展示肽库技术，可以成功筛选到F2，其可能成为喉癌靶向治疗的载体。【Abstract】　Objective　Toobtainthepolypeptidesspecificallyboundtolaryngealsquamouscellcarcinomaline(Hep-2)anduseitasapotentialtherapeuticvectortargetinglaryngealsquamouscellcarcinomapatients.　Methods　WiththeHep-2cellsasthetargetcellsandhumannormallaryngealsquamousepithelialcells(HNLEcells)astheabsorbercells,3roundsofpanningfromaPh.D.-12TMphage-displaypeptidelibrarywerecarriedout.Tenrandomlyselectedphageclonesweresentforsequencedetection.Theaffinityofphagecloneswasdetectedbyenzyme-linkedimmunosorbentassay(ELISA).Thepositivephageclones(F2)specificallyboundtoHep-2wereidentifiedbyimmunofluorescencedetection.　Results　After3roundsofscreening,5positivephageclonesshowedspecificbindingtoHep-2cellsandtheaffinityofpositivephageclones(F2)wassignificantlyhigherthanthatofthecontrolgroups(P<0.05).TheresultsofimmunofluorescencedetectionindicatedthatF2couldbespecificallyboundtoHep-2.　Conclusions　PhagedisplaypeptidelibrariestechniquecansuccessfullyscreenthepeptidespecificallyboundtoHep-2cellline.Thus,itprovidesapotentialvectorfortargetingtherapyoflaryngealsquamouscellcarcinomapatients.
%K 噬菌体展示
%K 喉癌细胞
%K 靶向性
%U http://www.hxyxqk.com.cn/oa/DArticle.aspx?type=view&id=201112023