人原发性肝癌相关抗原的蛋白质组学初步研究

发布时间：2018-02-02 06:52

本文关键词： 肝细胞癌蛋白组学相关抗原 Dna J同源B亚家族成员蛋白酶体亚基α　出处：《第三军医大学学报》2017年01期 　论文类型：期刊论文

【摘要】：目的检测肝细胞癌(hepatocellular carcinoma,HCC)的相关抗原,并予以验证。方法分别提取15例HCC组织及其癌旁组织总蛋白。Western blot分别检测各总蛋白与其HCC患者自体血清的杂交情况。双向电泳(two-dimensional electrophoresis,2-DE)技术将15例混合总蛋白中的各蛋白进行分离与染色。图像分析法比对染色胶上HCC组织及其癌旁组织蛋白的表达差异,基质辅助激光解吸/电离飞行时间二级质谱(matrix-assisted laser desorption/ionization-time-of-flight with two mass analyzers for tandem mass spectrometry,MALDI-TOF MS/MS)技术及生物信息学网络数据库分别对表达差异蛋白予以分析鉴定与检索匹配,免疫组织化学与Western blot分别检测该差异蛋白在人HCC组织及其癌旁组织中的变化情况。结果 2-DE染色胶图像分析比对结果显示,HCC及其癌旁组织蛋白表达差异斑点共90个,MALDI-TOF MS/MS与数据库对其中19个蛋白斑点分析鉴定与检索匹配发现15个蛋白表达上调,4个蛋白表达下调。免疫组织化学与Western blot检测结果显示,与对照比较,上调蛋白中的Dna J同源B亚家族成员11(dna J homolog subfamily B member 11,DNAJB11)在HCC组织中的表达明显增强,下调蛋白中的蛋白酶体亚基α3(proteasome subunit alpha type 3,PSMA3)在HCC组织中的表达则相反。结论人原发性肝癌相关抗原DNAJB11在HCC组织中的表达上调,PSMA3在HCC组织中的表达下调。
[Abstract]:Objective to detect the related antigens of hepatocellular carcinoma (HCC). Methods Total protein of 15 HCC tissues and their adjacent tissues were extracted. Blot was used to detect the hybridization of total proteins with autologous serum of patients with HCC. Two-dimensional electrophoresis. The proteins in 15 cases of mixed total proteins were separated and stained by 2-DE.The expression of proteins in HCC and its adjacent tissues were compared by image analysis. Matrix assisted laser desorption / ionization time of flight mass spectrometry. Matrix-assisted laser desorption/ionization-time-of-flight with two. Mass analyzers for tandem mass spectrometry. MALDI-TOF / MS / MS technique and bioinformatics network database were used to identify and search the differentially expressed proteins. Immunohistochemistry and Western blot were used to detect the changes of the differential protein in human HCC and its adjacent tissues. There were 90 differentially expressed spots in HCC and its adjacent tissues. The expression of 15 proteins was up-regulated by MALDI-TOF MS/MS and database. The expression of four proteins was down-regulated. The results of immunohistochemistry and Western blot showed that the protein expression was lower than that of control. Dna J homologous B subfamily member 11 DNA J homolog subfamily B member 11 was upregulated. The expression of DNA JB11 in HCC was significantly increased, and the proteasome subunit alpha type 3 was down-regulated. Conclusion the expression of human hepatocellular carcinoma associated antigen DNAJB11 in HCC is up-regulated and PSMA3 is down-regulated in HCC.
【作者单位】：重庆市肿瘤研究所血液肿瘤消化呼吸内科;重庆市肿瘤研究所血液肿瘤生物治疗科;
【基金】：重庆市应用开发计划项目(CSTC2014yykfA110031) 中国医师协会研究项目(ZGYSXH20120701)~~
【分类号】：R735.7
【正文快照】： identified by MALDI-TOF MS/MS,15 proteins were up-regulated,and 4 down-regulated.Immunohistochemicalassay and Western blotting confirmed that compared with the proteins from the adjacent tissues,Dna J homologsubfamily B member 11(DNAJB11)was significantl

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