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华西医学  2011 

用噬菌体展示技术筛选喉癌细胞靶向肽的研究

, PP. 1844-1847

Keywords: 噬菌体展示,喉癌细胞,靶向性

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Abstract:

【】 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.

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