CN106093429A - 一种检测胃癌组织的试剂盒 - Google Patents

一种检测胃癌组织的试剂盒 Download PDF

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CN106093429A
CN106093429A CN201610390597.4A CN201610390597A CN106093429A CN 106093429 A CN106093429 A CN 106093429A CN 201610390597 A CN201610390597 A CN 201610390597A CN 106093429 A CN106093429 A CN 106093429A
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test kit
stomach organization
antibody
prdx5
atp5a1
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王滨
田梗
米佳
李旭日
姜文国
位晓丹
刘芳
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Binzhou Medical College
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    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
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    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
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    • G01N33/57446—Specifically defined cancers of stomach or intestine
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere

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Abstract

本发明涉及一种检测胃癌组织的试剂盒，其特征在于，包含抗ATP5A1抗体和抗PRDX5抗体，所述ATP5A1的蛋白质序列如SEQ ID NO:1所示，所述PRDX5的蛋白质序列如SEQ ID NO:2所示。使用本发明，可有效地甄别正常组织和胃癌组织，既可用于科研工作中，也有助于胃癌早期诊断。

Description

一种检测胃癌组织的试剂盒
技术领域
本发明涉及肿瘤检测领域，更特别地，涉及一种检测胃癌组织的试剂盒。
背景技术
我国是胃癌高发国家，其发病率和死亡率分别高居全部恶性肿瘤的第3位和第2位。由于胃癌起病隐匿，早期多无明显症状，不易觉察，使得胃癌的早期发现非常困难。特别是在我国，超过80％的胃癌在确诊时就已处于进展期，预后很差，严重威胁着人们的生命健康。
氧化应激是指环境或细胞线粒体功能障碍等因素导致体内活性氧簇(reactiveoxygen species,ROS)如超氧阴离子、羟自由基、过氧化氢产生过多，氧化系统和抗氧化系统失衡，从而导致组织损伤。体内存在抗氧化酶如超氧化物歧化酶(superoxidedismutase，SOD)、过氧化氢酶(catalase,CAT)、谷胱甘肽过氧化物酶(glutathioneperoxidase，GST-Px)、过氧化物酶类(peroxiredoxins,PRDXs)等调节氧化应激反应。低浓度ROS可促进细胞增殖和分化，高浓度ROS则诱导细胞凋亡。
氧化应激和线粒体功能障碍之间有着紧密的相关联系。。线粒体是体内细胞功能和代谢方面起重要作用的细胞器，它的主要作用是产生ATP。线粒体的ATP合酶包含五个不同的亚基(α、β、γ、δ、ε)。ATP合酶广泛分布于线粒体内膜，参与氧化磷酸化，在跨膜质子动力势的推动下合成ATP。其中，ATP5A1和ATP5B基因分别负责编码533个氨基酸的α和529个氨基酸的β亚基。ATP5A1表达的增加可以增强ATP合酶的活性，ATP合酶活性的增强能减少线粒体超氧阴粒子的产生和氧化损伤。
氧化应激加重线粒体功能障碍，而线粒体功能障碍又增加ROS的产生，形成增强氧化应激的恶性循环。通过肿瘤线粒体蛋白质组分析，发现很多抗氧化损伤相关的蛋白(包括CAT、PRDX3和PRDX5等)表达增高。PRDXs是一类能降低ROS产生的抗氧化蛋白，能够增加细胞在氧化应激压力下存活和增殖能力，其表达水平与一些肿瘤如甲状腺癌、乳腺癌和肺癌相关。而PRDX5在对抗ROS方面比其他PRDXs更为有效，它能保护线粒体和核内DNA的损伤，降低其表达水平使得细胞易于凋亡。现在关于PRDX5和胃癌的关系，研究还非常少。
发明内容
发明人在研究胃癌的过程中，发现胃癌患者的胃癌组织样本中XBP1的表达受到显著抑制。在该发现的基础上，本发明提供了一种检测胃癌组织的试剂盒，其特征在于，包含抗ATP5A1抗体和抗PRDX5抗体，所述ATP5A1的蛋白质序列如SEQ ID NO:1所示，所述PRDX5的蛋白质序列如SEQ ID NO:2所示。
进一步地，所述试剂盒还包含针对所述抗ATP5A1抗体的具有第一显示标志物的二抗、与所述第一显示标志物对应的显示系统，以及针对所述抗PRDX5抗体的具有第二显示标志物的二抗、与所述第二显示标志物对应的显示系统。
进一步地，所述第一显示标志物和所述第二显示标志物都为辣根过氧化物酶。
进一步地，所述第一显示系统和所述第二显示系统独立地选自DAB显示系统或ECL显示系统。
进一步地，所述试剂盒还包含上样量标度。
进一步地，所述上样量标度为抗β-actin抗体。
使用本发明，可有效地甄别正常组织和胃癌组织，既可用于科研工作中，也有助于胃癌早期诊断。
附图说明
图1为使用本发明的试剂盒检测正常组织和胃癌组织中的ATP5A1蛋白含量的Westren blot图；
图2为使用本发明的试剂盒检测正常组织和胃癌组织中的PRDX5蛋白含量的Westren blot图。
具体实施方式
以下结合实施例和附图对本发明的原理和特征进行描述，所举实例只用于解释本发明，并非用于限定本发明的范围。
发明人在研究胃癌的过程中发现，ATP5A1和PRDX5的蛋白质水平在胃癌组织中都显示出显著降低，这提示，这两种蛋白的表达水平可用于鉴别胃癌组织和正常胃部组织。
实施例1检测胃癌组织的试剂盒
本实施例的试剂盒包含以下试剂：
1)抗ATP5A1抗体(ab176569，ABCAM)，辣根过氧化物酶标记的羊抗兔二抗；
2)抗PRDX5抗体(AF5724，R&G)，辣根过氧化物酶标记的兔抗羊二抗；
3)抗β-actin抗体及其二抗
4)DAB显示系统。
实施例2Western blot测定ATP5A1的表达
从正常组织和胃癌组织中提取总蛋白，进行SDS-PAGE电泳。使用蛋白质检测试剂组，按照常规的Western blot法，对总蛋白中的ATP5A1进行染色，结果如图1所示，胃癌组织(T泳道)中ATP5A1蛋白的含量显著高于正常组织(N泳道)。
实施例3Western blot测定PRDX5的表达
从正常组织和胃癌组织中提取总蛋白，进行SDS-PAGE电泳。使用蛋白质检测试剂组，按照常规的Western blot法，对总蛋白中的PRDX5进行染色，结果如图2所示，胃癌组织(T泳道)中PRDX5蛋白的含量显著高于正常组织(N泳道)。
以上所述仅为本发明的较佳实施例，并不用以限制本发明，凡在本发明的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。

Claims (6)

1.一种检测胃癌组织的试剂盒，其特征在于，包含抗ATP5A1抗体和抗PRDX5抗体，所述ATP5A1的蛋白质序列如SEQ ID NO:1所示，所述PRDX5的蛋白质序列如SEQ ID NO:2所示。
2.根据权利要求1所述的检测胃癌组织的试剂盒，其特征在于，还包含针对所述抗ATP5A1抗体的具有第一显示标志物的二抗、与所述第一显示标志物对应的显示系统，以及针对所述抗PRDX5抗体的具有第二显示标志物的二抗、与所述第二显示标志物对应的显示系统。
3.根据权利要求2所述的检测胃癌组织的试剂盒，其特征在于，所述第一显示标志物和所述第二显示标志物都为辣根过氧化物酶。
4.根据权利要求2所述的检测胃癌组织的试剂盒，其特征在于，所述第一显示系统和所述第二显示系统独立地选自DAB显示系统或ECL显示系统。
5.根据权利要求1所述的检测胃癌组织的试剂盒，其特征在于，还包含上样量标度。
6.根据权利要求5所述的检测胃癌组织的试剂盒，其特征在于，所述上样量标度为抗β-actin抗体。
CN201610390597.4A 2016-06-02 2016-06-02 一种检测胃癌组织的试剂盒 Pending CN106093429A (zh)

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Cited By (3)

* Cited by examiner, † Cited by third party
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CN106932597A (zh) * 2017-03-29 2017-07-07 山东大学 Atp5a1蛋白531位发生质量偏移的赖氨酸在制备重度少弱精诊断试剂中的用途
CN106932597B (zh) * 2017-03-29 2018-07-20 山东大学 Atp5a1蛋白531位发生质量偏移的赖氨酸在制备重度少弱精诊断试剂中的用途
WO2022013718A1 (pt) * 2020-07-13 2022-01-20 Universidade Federal De Minas Gerais Processo para classificação de amostras biológicas quanto a infecção por agentes virais e usos

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