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应用iTRAQ多重化学标记串联质谱技术筛选晚期卵巢浆液性腺癌组织中卡铂耐药相关蛋白[J]. 肿瘤防治研究, 2016, 43(1): 58-62. DOI: 10.3971/j.issn.1000-8578.2016.01.013
引用本文: 应用iTRAQ多重化学标记串联质谱技术筛选晚期卵巢浆液性腺癌组织中卡铂耐药相关蛋白[J]. 肿瘤防治研究, 2016, 43(1): 58-62. DOI: 10.3971/j.issn.1000-8578.2016.01.013
Screening of Potential Proteins Associated with Carboplatin-resistance in Advanced Ovarian Serous Adenocarcinoma Tissues Using iTRAQ Technology[J]. Cancer Research on Prevention and Treatment, 2016, 43(1): 58-62. DOI: 10.3971/j.issn.1000-8578.2016.01.013
Citation: Screening of Potential Proteins Associated with Carboplatin-resistance in Advanced Ovarian Serous Adenocarcinoma Tissues Using iTRAQ Technology[J]. Cancer Research on Prevention and Treatment, 2016, 43(1): 58-62. DOI: 10.3971/j.issn.1000-8578.2016.01.013

应用iTRAQ多重化学标记串联质谱技术筛选晚期卵巢浆液性腺癌组织中卡铂耐药相关蛋白

Screening of Potential Proteins Associated with Carboplatin-resistance in Advanced Ovarian Serous Adenocarcinoma Tissues Using iTRAQ Technology

  • 摘要: 目的 利用同位素标记相对和绝对定量(iTRAQ) 技术筛选晚期卵巢癌组织中卡铂耐药相关差异表达蛋白,为临床个体化治疗奠定实验基础。方法 收集Ⅲ期低分化卵巢浆液性腺癌标本并通过ATP-TCA药敏试验检测卡铂敏感度。取卡铂敏感和耐药标本各15例, iTRAQ试剂标记,被标记的肽段进行高效液相色谱(HPLC)分离及质谱检测(MS)。结果 共鉴定出iTRAQ标记定量信息有755个显著差异表达的蛋白。卡铂耐药组与卡铂敏感组相比,上调1.2倍以上的蛋白429个;下调0.83倍以下的蛋白326个。上调蛋白中有osteopontin(OPN)、clusterin(CLU)、5'-nucleotidase(CD73)和tissue inhititor of metalloproteinases 1(TIMP1)等18个蛋白与肿瘤恶性行为及化疗耐药相关。结论 应用iTRAQ技术能筛选出多种与晚期卵巢癌卡铂耐药相关的差异表达蛋白,该技术对于肿瘤组织蛋白质组学的研究有很好的应用前景。

     

    Abstract: Objective To screen the proteins associated with carboplatin-resistance in advanced ovarian serous adenocarcinoma tissues using proteomics approaches of isobaric tags for relative and absolute quantification(iTRAQ) to provide experimental basis for clinically individual treatment. Methods The specimens of primary phase Ⅲ ovarian serous adenocarcinoma tissues were collected and tested for the sensitivity to carboplatin using adenosine triphosphate tumor chemosensitivity assay (ATP-TCA). Then the samples from 15 carboplatin-resistant specimens and 15 carboplatin-sensitive specimens were mixed with equal weight. Protein samples subjected to enzymatic digestion of trypsin were labeled with identical iTRAQ tags respectively, and then the labeled peptides were separated and analyzed via high performance liquid chromatography (HPLC) followed by mass spectrometry (MS). Results A total of 755 differentially expressed proteins were identified. Compared with carboplatin-sensitive group, 429 proteins were significantly up-regulated (>1.2-fold) and 326 proteins were significantly down-regulated (<0.83-fold) in carboplatinresistant ovarian cancer tissues. Among these up-regulated proteins, 18 proteins were associated with tumor malignant behavior and chemoresistance, including Osteopontin (OPN), Clusterin (CLU), 5'-nucleotidase (CD73) and tissue inhibitor of metalloproteinases 1(TIMP1). Conclusion The differentially expressed proteins in carboplatin-resistant ovarian cancer tissues are identified by proteomic analysis using iTRAQ technology. The iTRAQ technology provides a good platform for the study of tumor proteomics.

     

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