文章摘要

利用蛋白质组学筛选肾透明细胞癌肿瘤标志物

作者: 1,2郭丹, 3王安琪, 3赵晓晓, 4刘辉, 1,5孙健
1 中国医学科学院北京协和医院临床生物标本中心，北京 100730
2 中国医学科学院北京协和医院中心实验室，北京 100730
3 中国医学科学院北京协和医院临床生物标本中心，北京 100730
4 中国医学科学院基础医学研究所，北京 100730
5 中国医学科学院北京协和医院病理科，北京 100730
通讯: 孙健 Email: sunjian0720@126.com
DOI: 10.3978/j.issn.2095-6959.2017.10.001

摘要

目的：通过蛋白质组学筛选和验证肾透明细胞癌中潜在肿瘤标志物。方法：采用比较蛋白质组学方法筛选正常肾组织和肾细胞癌(renal cell carcinoma，RCC)组织中的差异表达蛋白，通过二维差异凝胶电泳(two-dimensional difference gel electrophoresis，2D-DIGE)和傅里叶变换离子回旋共振质谱(Q-Fourier transform ion cyclotron resonance mass spectrometry，FT-ICR MS)分离并鉴定16例RCC组织及其相应的正常组织中差异显著的蛋白，并进一步通过实时荧光定量PCR(real-time quantitative PCR，qRT-PCR)、Western印迹和免疫组织化学在RNA和蛋白水平进行验证。结果：通过2D-DIGE共筛选出1 873个蛋白质点，选取68个表达差异大于1.5倍的蛋白质点进行鉴定，成功鉴定出51个蛋白质(P<0.05)，其中16个蛋白表达上调，35个表达下调。采用qRT-PCR和Western印迹验证16例肿瘤和对照组织，结果表明在肿瘤组织中NNMT和Annexin IV表达显著升高，而ACY1和Calbindin显著下调(P<0.01)。同时针对ACY1和Annexin IV蛋白分别在53例和47例样本中进行免疫组织化学，结果显示相较于对照组织，ACY1在肾透明细胞癌中表达显著降低(P<0.001)，而Annexin IV显著升高(P<0.001)。结论：ACY1和Annexin IV可作为肾透明细胞癌的肿瘤标志物。
关键词： 肾透明细胞癌 肿瘤标志物 蛋白质组学 二维差异凝胶电泳

Potential tumor markers of renal clear cell carcinoma with proteomic

Authors: 1,2GUO Dan, 3WANG Anqi, 3ZHAO Xiaoxiao, 4LIU Hui, 1,5SUN Jian
1 Clinical Biobank, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Beijing 100730
2 Corelabs, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Beijing 100730
3 Clinical Biobank, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Beijing 100730
4 Corelabs, Chinese Academy of Medical Sciences, Beijing 100730
5 Department of Pathology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Beijing 100730, China

CorrespondingAuthor: SUN Jian Email: sunjian0720@126.com

DOI: 10.3978/j.issn.2095-6959.2017.10.001

Abstract

Objective: To identify and validate the potential tumor markers of renal clear cell carcinoma for progress assessment and clinical treatment with proteomic. Methods: We analyzed 16 cases of tumor and pericarcinomatous tissue with two-dimensional difference gel electrophoresis (2D-DIGE) and Q-Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS), and then identified the abnormal expression protein with Western blot, Real-time quantitative PCR and IHC. Results: A total of 1 873 proteins were quantified with proteomic in 16 cases of renal clear cell carcinoma, and 51 proteins of 68 sports that more than 1.5-fold differential expressing were identified, including 16 proteins upregulated and 35 proteins downregulated in renal cell carcinoma (RCC) (P<0.05). Among them, the up-expressed NNMT, Annexin IV and down-expressed ACY1, calbindin in DIGE were confirmed with 16 cases of tumor and pericarcinomatous tissues with qRT-PCR and Western blot (P<0.01). Subsequently, SPSS analysis of IHC validated ACY1 decreased in 53 cases of clear cell renal cell carcinoma (P<0.001), and Annexin IV increased in 47 cases (P<0.001). Conclusion: ACY1 and Annexin IV serve as a putative tumor suppressor and a potential treatment marker for clear cell RCC.
Keywords： clear cell renal cell carcinoma tumor marker proteomics two-dimensional difference gel electrophoresis

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