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时间:2020-05-07
《应用免疫激活磁珠分选技术CD45去除方法富集——免疫细胞化学联合苏木素-伊红染色检测肝癌患者循环肿瘤细胞-论文.pdf》由会员上传分享,免费在线阅读,更多相关内容在应用文档-天天文库。
1、中华实验外科杂志2014年9月第31卷第9期ChinJExpSurg,September2014,Vo1.3l,No.9·2019··临床研究·应用免疫激活磁珠分选技术CD45去除方法富集——免疫细胞化学联合苏木素一伊红染色检测肝癌患者循环肿瘤细胞郭立民鲁岩彭吉润蒋力【摘要】目的探讨免疫激活磁珠分选(MACS)CIM5去除方法富集后,以细胞角蛋白(CK)为标记联合苏木素一伊红(HE)染色检测肝癌患者外周血肿瘤细胞(CTC)的价值。方法应用逆转录一聚合酶链反应(RT.PCR)检测18种上皮肿瘤细胞株的CK(CK8、CK18、CK19)表达;采集健康志愿者、肝
2、癌患者外周血,以MACS技术CD45去除方法对外周血单个核细胞(PBMC)进行富集,以CK为标记,采用免疫组织化学染色联合HE染色检测肿瘤细胞。结果在18种上皮性肿瘤细胞株中CK8、CK18、CK19表达率分别为72.22%、83.33%和66.67%;9种肝癌细胞株中CK阳性表达占绝对多数;在MACS技术CD45去除富集肿瘤细胞基础上,应用免疫细胞化学和HE染色结果显示,肝癌患者外周血存在完整肿瘤细胞,检出率为63.15%(12/19),健康志愿者未检测到肿瘤细胞。结论CK可作为检测外周血肝癌细胞标志物;MACS技术CD45去除方法富集后,CK免疫细胞化
3、学和HE染色同步检测方法有助于检测肝癌患者外周血肿瘤细胞。【关键词】循环肿瘤细胞;细胞角蛋白;免疫激活磁珠分选;苏木素一伊红染色Combineduseofimmunomagneticactivatedcellsortingtechniqueenrichmentandimmunocyto-chemistrywithhematoxylinandeosinstainingforidentificationofcirculatingtumorcellsinperiph-eralbloodmononuclearcellsofhepatocellularcarcino
4、mapatientsGuoLimin}J1lYan.PmgJirua。JiangLi.DepartmentofSurgery,BeijingDitanHospital,CapitalMedicalUniversity,Beijing1oo0l5.ChinaCorrespondingauthor:JiangLi,Emaif:movdush@139.com【Abstract】ObjectiveToestimatetheappliedvalueofmagneticactivatedcellsorting(MACS)techniqueswithCD45deplet
5、ionandimmunocytochernistryincombinationwithhematoxylinandeosin(HE)staininginidentifyingcirculatingtumorcells(CTCs)inperipheralbloodmononuclearcells(PBMC)ofhep—atocellularcarcinoma(HCC)patients.MethodsTheexpressionofCK(CK8,CK18,andCK19)wasde—tectedin18epithelia.derivedtumorcellline
6、sincluding9humanhepatocellularcarcinomacelllines.Thepe—ripheralbloodofHCCpatientsandheMthyvolunteerswascollectedfordeterminationofCTCsinPBMCfromHCCpatientsusingMACStechniqueswithCD45depletionandimmunocytochemstryincombinationwithHEstaining.ResultsTheexpressionrateofCK8.CK18andCK19
7、intheselectedCTCswas72.22%.83.33%and66.67%respectively.CKsweredetectedinmostofthe9hepatocelluarcarcinomacelllines.WefoundintactCTCsinPBMCfromHCCpatientsusingHEstainingandimmunocytochemistryafterPBMCenrichmentbvMACStechniqueswithCD45depletion.Thesensitivityofthismethodwasupto63.15%
8、.andnoCTCsweredetectedinPBMCfrom2
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