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SLC35F2抗体制备与非小细胞肺癌组织中SLC35F2蛋白表达检测
  • ISSN号:1001-9030
  • 期刊名称:中华实验外科杂志
  • 时间:0
  • 页码:1046-1048
  • 语言:中文
  • 分类:R373.2[医药卫生—病原生物学;医药卫生—基础医学] R734.2[医药卫生—肿瘤;医药卫生—临床医学]
  • 作者机构:[1]北京大学人民医院胸外科,100044
  • 相关基金:国家自然科学基金资助项目(30672084)
  • 相关项目:肺癌相关新基因LSCC-3编码蛋白SLC35F2(溶质载体35家族F2成员)的结构、特异性底物及其功能的进一步研究
作者: 刘军|
中文摘要:

目的制备SLC35F2多肽抗体,鉴定其性能,检测其对应蛋白在非小细胞肺癌(NSCLC)组织中的表达。方法通过蛋白质序列分析,选4段氨基酸序列合成多肽,混合免疫家兔,制备抗SLC35F2多肽抗体,经纯化、效价检测、特异性鉴定后,将抗体用于NSCLC标本免疫组织化学染色。结果获得兔抗入SLC35F2多肽抗体,效价1:10^5,Westernblot可特异识别41KD的SLC35F2蛋白,免疫组织化学示细胞胞质特异性染色。129例NSCLC的组织微阵列免疫组织化学染色显示,SLC35F2蛋白表达在90.7%(117/129)肺癌标本为廿到卅;而仅在17.1%(22/129)癌旁正常肺组织为廿,其余为-或+;SLC35F2蛋白表达在NSCLC组织中高于癌旁正常肺组织(P〈0.01);NSCLC病理分期与SLC35F2的升高表达存在低度相关(r=0.175)。结论用多肽免疫家兔制备抗SLC35F2抗体具有较高的效价和特异性,SLC35F2在NSCLC中表达高于癌旁正常组织。

英文摘要:

Objective To prepare, purify and characterize the polyclonal antibody against SLC35F2, and detect the expression of SLC35F2 protein in non-small-cell lung cancer (NSCLC). Methods Four polypeptides named peptide 1,2,3 and 4 were synthesized based on the bioinfformatics analysis of SLC35F2. The mixed polypeptides were injected into New Zealand rabbits. The polyclonal antibody was purified by immunoaffinity chromatography, and identified by Western blot and immunohistochemistry. The expression of SLC35F2 protein in 129 cases of NSCLC and corresponding adjacent normal lung tissues was detected by tissue microarray with immunohistochemistry. Results The titers of polyclonal antibody were 1:10^5 detected by ELISA assay. Western blot confirmed its high specific 41 KD band. The expression of SLC35F2 protein was strongly positive ( ++ to +++ ) in 90.7% (117/129) of the NSCLC samples. On the contrast ,only 17.1% (22/129) adjacent normal lung tissues were strongly positive ( ++ ). The expression of SLC35F2 protein was significantly higher in NSCLC than in the adjacent normal lung tissues ( P 〈 0.01 ). Correlation analysis revealed that pathological stage had a low correlation with the expression of SLC35F2 protein. Factor analyses showed that 93.3% expression of SLC35F2 protein in NSCLC tissues were correlated with tumor differentiation and stage. Conclusion The specific antibody against human SLC35F2 was obtained. SLC35F2 protein was highly expressed in most of the NSCLC.

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期刊信息
  • 《中华实验外科杂志》
  • 北大核心期刊(2011版)
  • 主管单位:中国科学技术协会
  • 主办单位:中华医学会
  • 主编:
  • 地址:武汉市武昌区东湖路165号
  • 邮编:430064
  • 邮箱:cjes@cma.org.cn
  • 电话:027-87893475
  • 国际标准刊号:ISSN:1001-9030
  • 国内统一刊号:ISSN:42-1213/R
  • 邮发代号:38-85
  • 获奖情况:
  • 国内外数据库收录:
  • 美国化学文摘(网络版),日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),中国北大核心期刊(2000版)
  • 被引量:34996