基础医学与临床 ›› 2012, Vol. 32 ›› Issue (2): 191-194.

• 研究论文 • 上一篇    下一篇

宫颈癌血清相关蛋白的初步探讨及鉴定

杨佳欣1,郎景和1,沈铿2,曹冬焱1,荣春红3   

  1. 1. 北京协和医院妇产科
    2. 北京协和医院妇产科 北京
    3. 中日友好医院妇产科
  • 收稿日期:2009-12-02 修回日期:2011-04-07 出版日期:2012-02-05 发布日期:2012-01-12
  • 通讯作者: 杨佳欣 E-mail:yangjiaxin007@hotmail.com
  • 基金资助:
    院校级基金

Primary research and identification of associated proteins in serum of cervical cancer

  • Received:2009-12-02 Revised:2011-04-07 Online:2012-02-05 Published:2012-01-12

摘要: [摘要] 目的:寻找宫颈癌患者和对照组之间血清的蛋白差异,分离纯化差异蛋白,并以质谱技术进行鉴定。方法:SELDI-TOF-MS分析宫颈癌患者和对照组血清蛋白表达得到差异蛋白/肽,离子交换层析、SDS-PAGE进一步分离纯化差异蛋白,SELDI-TOF-MS在分离纯化过程中验证差异蛋白的存在,最后ESI-LC-MS/MS对差异蛋白进行质谱鉴定。结果:宫颈癌患者和对照组血清之间差异蛋白( P<0.05)12个,对其中M/Z 15960Da的差异蛋白进行分离纯化鉴定,比对数据库鉴定出8类蛋白,与15960Da分子量最接近的是甲状腺激素转运蛋白( TTR),有少数研究表明它的某些修饰形式与肿瘤有关。从蛋白的生物学行为上最有意义的是转录中介因子TIF1-β,也叫做KAP-1/KRAB/TRIM28,它的表达可能引起肿瘤相关基因的失活或活化,导致肿瘤发生发展。结论:我们在国内首次采用SELDI-TOF-MS寻找差异蛋白并辅助蛋白分离纯化,LC-ESI-MS/MS鉴定蛋白的技术路线。对M/Z15960Da的差异蛋白分离纯化鉴定,数据库匹配出8类可能的蛋白,TTR的分子量最接近,而TIF1-β生物学行为意义更重要。

关键词: 宫颈癌 SELDI-TOF-MS TIF1-β TTR

Abstract: Primary research and identification of associated proteins in serum of cervical cancer Rong Chun-hong, Shen Keng, Yang Jia-xin, .Department of Obstetrics and Gynecology Peking Union Medical College Hospital,Beijing,100730,China [Abstract] Objective: To discover different proteins between cervical cancer and control group , then we separate, purify and enrich the interest proteins and identify them by MS. Methods We use SELDI-TOF-MS to analyze the difference peptide profiling and proteins between serum of cervical cancer patients and the control group, interested proteins are subsequently separated and enriched by ion chromatographic fractionation, SDS-PAGE ,at last the proteins are identified by LC-MS/MS analysis, SELDI-TOF-MS is used to verify the existence of interest proteins. Results 12 different proteins are identified in serum between cervical cancer and the control group(P<0.05). We separate, purify and identify interest proteins whose M/Z is 15960Da and get 8 kinds of proteins according the database. The most adjacent to 15960Da molecular weight comes from transthyretin (TTR), which there are some subjects showed it’s correlation to tumor. The TIF1-β protein, namely KAP-1/KRAB/TRIM28, arose us by it’s biologic action, because it’s expression can inhibit or activate tumor related genes then promote the development of malignant tumor. Conclusions We are the first in China who applying the technology course that discover different proteins by SELDI-TOF-MS , purify and enrich interested proteins monitoring by SELDI-TOF-MS, identify it by LC-ESI-MS/MS. We purify and identify the protein whose M/Z is 15960Da, data of IPI gives us 8 kinds of possible proteins, in which the molecule weight of TTR is the most close, but TIF-1β has the most important biologic behavior . [Key words] cervical cancer SELDI-TOF-MS TIF1-β TTR

Key words: cervical cancer SELDI-TOF-MS TIF1-β TTR

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