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目的:通过比较早期宫颈癌和中-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-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Western Blot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-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-晚期组下调的蛋白4个,包括Hemoglobin subunit beta HB、caspase-14、galectin-7、CK19;中-晚期组上调的蛋白8个,包括NMP238、HSP70、Calmodulin-like5、S100A9。Western Blot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。
OBJECTIVE: To identify differentially expressed proteins in early stage cervical cancer and metaphase-advanced stage cervical cancer and to find out the proteins related to clinical staging of cervical cancer so as to provide new indexes for predicting clinical prognosis and recurrence. Methods: The specimens of early stage cervical cancer and middle-late stage cervical cancer were collected, and the total tissue proteins were extracted for two-dimensional gel electrophoresis. Some differentially expressed proteins were selected for MALDI-TOF mass spectrometry and bioinformatics analysis for protein identification. Western Blot And immunohistochemistry to detect the expression of some differentially expressed proteins. Results: The two-dimensional gel electrophoresis patterns of early stage cervical cancer group and middle-late stage cervical cancer group were established. The average protein spots of the two groups were 1098 ± 23,1142 ± 21, respectively. Through mass spectrometry and bioinformatics query, Compared with the early cervical cancer group, four proteins were identified that were down-regulated in the metaphase-late group, including Hemoglobin subunit beta HB, caspase-14, galectin-7 and CK19; , Calmodulin-like5, S100A9. The results of Western Blot and immunohistochemistry showed that the expression of S100A9 in the middle-late stage group was higher than that in the early stage group. The expression of CK19 in the early stage group was higher than that in the middle-late stage group. Conclusion: There are differentially expressed proteins in early cervical cancer and middle-late cervical cancer. These proteins may be biomarkers for prognosis and recurrence of cervical cancer.