宫颈上皮内瘤变与宫颈鳞癌组织的差异蛋白质组学研究

Differential Proteomics of Cervical Intraepithelial Neoplasia and Squamous Cervical Carcinoma Tissue

[目的]比较宫颈上皮内瘤变(CIN)与宫颈鳞癌(SCC)组织之间蛋白质组的差异,寻找与宫颈癌相关的特异性蛋白质,为研究CIN发展为宫颈癌的分子机制及临床诊治工作提供新的线索。[方法]收集正常宫颈组织9例、CIN组织23例(其中CINⅠ7例、CINⅡ8例、CINⅢ8例)及SCC组织7例。应用二维荧光差异凝胶电泳(2-DDIGE)寻找差异表达蛋白质点,基质辅助激光解析飞行时间串联质谱(MALDI-TOF/TOFMS)分析差异蛋白质点。进一步对3个重要差异蛋白S100A9、eEF1A1及PKM2应用免疫组织化学定性及WesternBlot定量检测其在宫颈组织中的表达。[结果]建立了分辨率高、重复性好的CIN与SCC的双向凝胶电泳图谱。两组差异蛋白质点共860个,标记蛋白质点46个(27个上调,19个下调),成功鉴定25个蛋白质。免疫组化及免疫印迹结果显示S100A9在SCC中表达水平高于CIN,而PKM2、eEF1A1在SCC中表达水平显著低于CIN。[结论]CIN与SCC间存在蛋白质表达的差异,这些差异表达的蛋白质可能成为宫颈癌早期诊断的标志物及治疗的新靶点。

[Purpose] To identify candidate biomarkers for cervical cancer and reveal related molecular mechanism for clinical diagnosis and treatment. [Methods] Tissue samples from pa- tients with squamous cervical carcinoma （SCC） （7 cases） , cervical intraepithelial neoplasia （CIN） （23 cases） and dlsease-free controls （9 cases） were collected. 2-D DIGE was applied to separate proteins and the images of the gels were analyzed. Then MALDI-TOF/TOF MS was used to identi- fy the differentially expressed proteins and further to validate 3 strikingly differential proteins SIOOA9,eEFIAI and PKM2 with immunohistochemistry and western blot. [Results] A total of 46 differential proteins-spots （19 up-regulated and 27 down-regulated） were marked between CIN and SCC, of which 25 proteins were identified. Immunohistochemistry and Western Blot confirmed that the expression level of S100A9 was higher in SCC than that in CIN,while PKM2 and eEF1AI significantly down-regulated in SCC compared with those in CIN. [Conclusion] The differentially expressed proteins between CIN and SCC are potentially candidate markers for early diagnosis of cervical cancer .