抗RecQ解旋酶单克隆抗体6H5在几种肿瘤细胞中的初步应用  

Preliminary Application of Monoclonal Antibody 6H5 Anti-RecQ Helicase in Several Tumor Cells

作　　者：张荣红[1,2] 廉芳 周孟[3] 贾铁文 何志旭[1] 刘杰麟[1,2] ZHANG Ronghong;LIAN Fang;ZHOU Meng;JIA Tiewen;HE Zhixu;LIU Jielin(Tissue Engineering and Stem Cell Researeh Center,Guizhou Medical University,Guiyang 550004,Guizhou,China;Department of Immunology,Guizhou Medical University,Guiyang 550004,Guizhou,China;Engineering Researeh Center for the Development and Application of Ethnic Medicine and TCM,Ministry of Education ＆ State Key Laboratory of Functions and Applications of Medicinal Plants,Guizhou Medical University,Guiyang 550004,Guizhou,China)

机构地区：[1]贵州医科大学组织工程与干细胞实验中心,贵州贵阳550004 [2]贵州医科大学免疫学教研室,贵州贵阳550004 [3]贵州医科大学民族药与中药开发应用教育部工程研究中心国家苗药工程技术研究中心,贵州贵阳550004

出　　处：《贵州医科大学学报》2018年第10期1209-1216,共8页Journal of Guizhou Medical University

基　　金：国家自然科学基金资助项目(81360349);贵州省优秀科技教育人才省长专项资金资助项目[黔省专合字(2006)57];贵州省国际科技合作计划基金资助项目[黔科合外G字(2011)7012];贵州省应用基础研究计划重大专项子课题资助项目[黔科合J重大字(2015)2003]

摘　　要：目的:制备抗RecQ解旋酶单克隆抗体(6H5抗体),鉴定其生物学特性并开展其对肿瘤细胞及肿瘤干细胞中RecQ解旋酶的检测。方法:在Balb/c小鼠腹腔接种杂交瘤细胞6H5后收集腹水,ELISA法鉴定腹水中6H5抗体,间接免疫荧光技术检测6H5抗体与乳腺癌细胞MDA-MB-435、人肾癌细胞GRC-1、人舌癌细胞Tca8113中BLM、RecQ4和RecQ5的结合;以6H5抗体作为一抗,免疫组织化学染色法分析Jurkat、MDA-MB-435肿瘤细胞及肿瘤干细胞中RecQ解旋酶的表达水平。结果:经鉴定,小鼠腹水中6H5抗体阳性,该抗体能识别不同肿瘤细胞株中的BLM、RecQ4和RecQ5解旋酶,还可敏锐检测Jurkat、MDA-MB-435肿瘤细胞与肿瘤干细胞中RecQ解旋酶的表达; RecQ解旋酶在Jurkat肿瘤干细胞中表达水平明显高于Jurkat肿瘤细胞,而在MDA-MB-435肿瘤干细胞表达水平则低于MDA-MB-435肿瘤细胞,差异有统计学意义(P <0. 05)。结论:RecQ家族解旋酶可以作为多种肿瘤细胞及肿瘤干细胞表面标志物的备选指标。Objective： To prepare monoclonal antibodies （6H5） against RecQ helicase, to identify its biological characteristics and carry out detection of RecQ helicase in tumor stem ceils. Methods： Inoculating hybridoma cell 6H5 in abdomen of Balb/c rats and then collecting ascitic fluid; adopting ELISA assay to identify 6H5 antibody in the fluid, and indirect immunofluorescence to detect the effects of 6H5 antibody and breast cancer cell MDA-MB-435, GRC-1 of human renal cancer cells, BLM, RecQ4 and RecQ5 in Tca8113 of human tongue cancer cells. Immunohistochemical method was used to analyze RecQ helicase expression level in Jurkat, breast cancer cell MDA-MB-435 and its stem cells. Results： 6H5 antibody in ascitic fluid of rats abdomen was positive, and such antibody could identify- BLM, RecQ4 and RecQ5 helicase in different tumor cells, and can also detect the expression of RecQ helicase expression in Jurkat, MDA-MB-435 tumor cells and tumor stem cells. RecQ helicase expression level in Jurkat tumor stem cell was obviously higher than Jurkat stem cell, while its expression in MDA-MB-435 tumor cells was lower than MDA-MB-435 tumor stem cell, difference was statistically significant （P 〈 0.05 ）. Conclusion ： RecQ family helicase can be used as an ahernative marker for tumor cells and tumor stem cell surface markers.

关 键 词：RecQ解旋酶 单克隆抗体 免疫荧光 免疫组织化学染色 肿瘤 

分 类 号：R730[医药卫生—肿瘤]