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小鼠肝癌细胞H22膜蛋白组的双向电泳
  • ISSN号:1007-3418
  • 期刊名称:《中华肝脏病杂志》
  • 时间:0
  • 分类:R73-35[医药卫生—肿瘤;医药卫生—临床医学] R734.2[医药卫生—肿瘤;医药卫生—临床医学]
  • 作者机构:[1]泸州医学院医学分子生物学实验室,646000
  • 相关基金:国家自然科学基金(70071040);泸州医学院科研基金(200601)
中文摘要:

目的运用双向电泳及生物信息学技术,对小鼠肝癌细胞H22与正常肝细胞的膜垡白进行比较和鉴定,以期为探索膜蛋白在肝癌发生、发展及侵袭转移过程中的作用提供实验依据。方法运用顺序抽提法结合与双向电泳兼容的膜蛋白抽提试剂盒提取小鼠肝癌细胞H22和小常小鼠肝细胞膜蛋白,用考马斯亮蓝法定量,经舣向凝胶电泳分离、银染,Image Master 2D Platinum软件分析,切取差异蛋白点,采用蔡质辅助激光解吸附飞行时间质谱技术检测,Aldente软件检索Swiss-Prot数据库进行匹配鉴定。结果与正常小鼠肝纠胞膜蛋白双向电泳图谱对比分析鉴定了8个在小鼠肝癌细胞H22中表达明晁增强的膜赁白,分别为硫酸酯酶修饰因子2,蛋白激酶C和肌酸激酶Ⅱ底物蛋白3,序列装配结构组件50同系物,巨噬细胞清道人受体Ⅰ/Ⅱ,非特异生白C9或f135同系物,紧密结合蛋白ZO-2,3羟基3甲基戊二酰辅酶A还原酶,空泡蛋由序列相关蛋白52同系物。结论已鉴定的8个膜蛋白的生理功能涉及细胞代谢、细胞增殖、细胞信号转导、细胞骨架等,且在肝癌细胞H22中表达明显增强,提示小鼠肝癌细胞H22发牛发聪及侵袭转移可能涉及相关膜蛋白生理功能的改变。

英文摘要:

Objective To compare the membrane protein profile of mouse hepatocarcinoma cell H22 with that of normal liver cell. Methods The membrane proteins in mouse hepatocarcinoma cell H22 and normal liver cell were extracted and their concentrations were determined by Bradford method. The proteins were separated by two-dimensional electrophoresis, and then stained with silver. The 2-DE maps were scanned and analyzed by Image Master 2D Platinum software. The differential expression protein spots were cut out from the gels, and the peptide fingerprinting was determined by MALDI-TOF-MS (Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry), followed by matching to Swiss-Prot protein database by Aldente software with experimental pI and MW data. Results Compared to normal liver cells, 8 membrane proteins, including sulfatase-modifying factor 2, protein kinase C and casein kinase II substrate protein 3, sorting and assembly machinery component 50 homolog, macrophage scavenger receptor types Ⅰ/Ⅱ, uncharacterized protein C9 or f135 homolog, tight junction protein ZO-2, 3-hydroxy-3- methylglutarylcoenzyme A reductase, and vacuolar protein sorting-associated protein 52 homolog were upregulated in H22 cells. Conclusion The membrane proteins involved in cell metabolism, proliferation, signal transduction, and skeleton, which are highly expressed in mouse hepatocarcinoma H22 cells, are probably related to the proliferation, invasion and migration of this tumor cell line.

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期刊信息
  • 《中华肝脏病杂志》
  • 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:重庆市渝中区临江路74号
  • 邮编:400010
  • 邮箱:chnhepa@online.cq.cn
  • 电话:023-63706512
  • 国际标准刊号:ISSN:1007-3418
  • 国内统一刊号:ISSN:50-1113/R
  • 邮发代号:78-56
  • 获奖情况:
  • 中国期刊方阵“双效”期刊
  • 国内外数据库收录:
  • 美国化学文摘(网络版),荷兰文摘与引文数据库,美国生物医学检索系统,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)
  • 被引量:47128