一种优化改良的胆汁蛋白质组学研究方法
| 作者: |
1柏强善,
1阴继凯,
1袁利娟,
1王成果,
1杨媛,
1臧莉,
1孙艺波,
2李泽,
2乔健,
1王青,
1鲁建国
1 第四军医大学唐都医院 普通外科，陕西 西安 710038 2 第四军医大学 学员旅，陕西 西安 710032 |
| 通讯: |
鲁建国
Email: lujguo@hotmail.com |
| DOI: | 10.3978/.2018.02.009 |
| 基金: | 陕西省科技统筹创新工程计划基金资助项目（2015KTCL03-05）。 |
摘要
目的：探索一种可靠、方便、高通量的胆汁蛋白质组学研究的方法。
方法：抽取3例胆囊结石和3例胆囊癌患者的胆囊胆汁进行蛋白质提取纯化和定量，然后利用高通量的蛋白质组学研究方法同位素标记相对和绝对定量（iTRAQ）进行蛋白质的鉴定以及定量，并进行生物信息学分析。
结果：经浓度和完整性检测，提取的胆汁蛋白质样品的浓度和完整性均达到了iTRAQ实验的要求。经过鉴定，共检测出1 323种蛋白质，比传统方法检测出的蛋白质数量显著提高。与结石患者胆汁比较，肿瘤患者胆汁有173种蛋白质明显上调，有345种蛋白质明显下调（变化倍数>1.5，P<0.05）。
结论：建立了一种可靠的胆汁蛋白质分离、纯化、鉴定、分析的方法，鉴定出的蛋白质数量比传统方法明显增多，更有潜力发现与疾病相关的蛋白质分子，为日后胆汁以及其他体液的蛋白质组学研究奠定了基础。
关键词：
胆囊肿瘤；胆汁；蛋白质组学；计算生物学
方法：抽取3例胆囊结石和3例胆囊癌患者的胆囊胆汁进行蛋白质提取纯化和定量，然后利用高通量的蛋白质组学研究方法同位素标记相对和绝对定量（iTRAQ）进行蛋白质的鉴定以及定量，并进行生物信息学分析。
结果：经浓度和完整性检测，提取的胆汁蛋白质样品的浓度和完整性均达到了iTRAQ实验的要求。经过鉴定，共检测出1 323种蛋白质，比传统方法检测出的蛋白质数量显著提高。与结石患者胆汁比较，肿瘤患者胆汁有173种蛋白质明显上调，有345种蛋白质明显下调（变化倍数>1.5，P<0.05）。
结论：建立了一种可靠的胆汁蛋白质分离、纯化、鉴定、分析的方法，鉴定出的蛋白质数量比传统方法明显增多，更有潜力发现与疾病相关的蛋白质分子，为日后胆汁以及其他体液的蛋白质组学研究奠定了基础。
An optimized and improved method for bile proteomic analysis
CorrespondingAuthor:LU Jianguo Email: lujguo@hotmail.com
Abstract
Objective: To establish a reliable, convenient and high resolution method for bile proteomic analysis.
Methods: Gallbladder bile samples were drawn from 3 gallbladder cancer patients and 3 gallstone patients for protein extraction and purification, and then the protein samples were identified and quantified using a high-resolution proteomic method named isobaric tags for relative and absolute quantification (iTRAQ), and then were analyzed by bioinformatics methods.
Results: After concentration and integrity test, all of the extracted protein samples met the concentration and integrity requirements of the iTRAQ experiment. Following identification, a total of 1 323 proteins were detected, which was remarkably higher than that by traditional methods. In bile from gallbladder cancer patients compared with that from gallstone patients, 173 proteins were significantly up-regulated and 345 proteins were significantly down-regulated (fold change>1.5, P<0.05).
Conclusion: A reliable method for bile protein isolation, purification, identification and analysis is established, which has the ability to detect many more proteins and greater potential to find key proteins associated with specific diseases compared with traditional methods, and may lay a foundation for proteomic analysis of bile and other body fluids in the future.
Keywords：
Gallbladder Neoplasms; Bile; Proteomics; Computational Biology
Methods: Gallbladder bile samples were drawn from 3 gallbladder cancer patients and 3 gallstone patients for protein extraction and purification, and then the protein samples were identified and quantified using a high-resolution proteomic method named isobaric tags for relative and absolute quantification (iTRAQ), and then were analyzed by bioinformatics methods.
Results: After concentration and integrity test, all of the extracted protein samples met the concentration and integrity requirements of the iTRAQ experiment. Following identification, a total of 1 323 proteins were detected, which was remarkably higher than that by traditional methods. In bile from gallbladder cancer patients compared with that from gallstone patients, 173 proteins were significantly up-regulated and 345 proteins were significantly down-regulated (fold change>1.5, P<0.05).
Conclusion: A reliable method for bile protein isolation, purification, identification and analysis is established, which has the ability to detect many more proteins and greater potential to find key proteins associated with specific diseases compared with traditional methods, and may lay a foundation for proteomic analysis of bile and other body fluids in the future.