应用蛋白质组学技术筛选宫颈癌临床分期相关蛋白

目的：通过比较早期宫颈癌和中一晚期宫颈癌的差异表达蛋白，以发现与宫颈癌临床分期相关蛋白，为临床预后和复发的预测提供新指标。方法：收集早期宫颈癌和中一晚期宫颈癌组织标本，提取组织总蛋白进行二维凝胶电泳，选择部分差异表达蛋白进行MALDI—TOF质谱分析和生物信息学分析进行蛋白质鉴定，进一步应用WestemBlot和免疫组织化学技术检测部分差异表达蛋白的表达情况。结果：建立了早期宫颈癌组和中一晚期宫颈癌组的二维凝胶电泳图谱，其中两组的平均蛋白质点数分别为1098±23、1142±21，通过进行质谱分析和生物信息学查询，与早期宫颈癌组比较，鉴定了在中一晚期组下调的蛋白4个。包括HemoglobinsubunitbetaHB、caspase-14、galectin-7、CK19；中-晚期组上调的蛋白8个，包括NMP238、HSP70、Calmodulin-like5、S100A9。WesternBlot和免疫组化检测结果均显示S100A9在中-晚期组中的表达高于早期组，CK19在早期组中的表达高于中-晚期组。结论：早期宫颈癌和中-晚期宫颈癌中存在着差异表达蛋白，这些蛋白可能成为宫颈癌预后和复发预测的生物标志物。

Screening of Clinical Stage associated proteins in cervical carcinoma by Proteomic technique

Objective： Cervical carcinoma clinical stage associated proteins would be found by comparing differential expressed proteins between early stage cervical carcinoma and medium-advanced stage cervical carcinoma, in order to find some new indexes for predicting the prognosis and recurrence. Methods： Cervical carcinoma tissues with different clinical stages were collected respectively. Total protein from cervical carcinoma tissues were extracted; differential proteome profiles were established and analyzed by means of two-dimensional polyacrylamide gel electrophoresis （2D-PAGE）, matrix-assisted laser desorption/ionization time of flight mass speetrometry （MALDI-TOF-MS） and bioinformatics analysis. Part of proteins expression were assayed by Western Blot and immunohistochemical method. Results： Well-resolved, reproducible 2-DE profiles of human cervical carcinoma tissues were obtained. Average protein dots were 1098± 23, 1142± 21 in early-stage and medium-advanced stage respectively; By analyzing and by mass spectrometry and bioinformatics, 8 of them were well characterized. Compared to early stage group, 4 proteins were low-expressed in early-stage group, includirig Hemoglobin subunit beta HB, caspase-14,galectin-7,CK19; 4 protein were highly expressed in medium-advanced stage, including NMP238, HSP70, Calmodulin-like 5,S100A9. S100A9 expressed higher in medium-advanced stage group than in early-stage group and CK expressed higher in early-stage stage group than in medium-advanced group. Conclusions： Differential expressed proteins exist between early-stage cervical carcinoma and medium-advanced cervical carcinoma, which would be biomarkers for prediction of prognosis and recurrence.