胆囊癌组织蛋白质组研究及膜联蛋白A3表达的临床病理意义

Proteomic study of gallbladder cancer, with special reference on the expression and significance of annexin A3

目的 通过分离并鉴定胆囊癌和胆囊良性组织的差异表达蛋白质,以发现可能用于早期诊断或治疗的胆囊癌肿瘤标志物.方法 提取人胆囊癌和胆囊良性组织（各6例）的总蛋白质,用双向电泳分离蛋白并进行比较.选择在胆囊癌组织中明显差异表达的蛋白点,行质谱分析.采用免疫组织化学EliVision法,对50例原发性胆囊癌38例慢性胆囊炎组织中膜联蛋白A3表达进行检测,分析胆囊癌中膜联蛋白A3的表达状况及其与胆囊癌临床病理指标和预后的关系.结果 获得了分辨率和重复性均很好的凝胶蛋白图谱.对筛选出的在胆囊癌组织中明显差异表达的46个蛋白点,共有17种蛋白质被成功鉴定,其中在胆囊癌组织中高表达9个,低表达8个.膜联蛋白A3在胆囊癌组织中表达的阳性率明显高于慢性胆囊炎组织,差异有统计学意义（74.0%：21.1%,P〈0.01）,其表达水平与胆囊癌患者年龄、性别及组织学类型等因素均无明显关系（P〉0.05）,但高表达与胆囊癌的低分级（40.0%：82.5%,P〈0.05）、淋巴结或远处转移（40.9%：100.0%,P〈0.05）以及术后生存时间减少（50.0%：93.8%,P〈0.05）有明显关系.结论 胆囊癌组织蛋白与胆囊良性组织存在明显的差异.膜联蛋白A3可能对胆囊癌的发生、发展有作用.

Objective To explore the potential molecular targets for diagnosis and treatment of gallbladder cancer by analyzing and comparing the proteomes expressed in human gallbladder cancer and benign gallbladder tissues. Methods The proteins expressed were analyzed using two-dimensional gel electrophoresis. The differentially expressed proteins in tumors were also analyzed by mass spectrometry （MS）. AnnexinA3 expression was examined by streptavidin peroxidase immunohistochemical technique on paraffin-embedded tissue sections from SO patients of gallbladder cancer and 38 cases of chronic eholecystitis. Results Protein extracts of individual sample in each type of tissues were separated on two-dimensional gels. There were forty six differentially expressed proteins in the tissue samples of gallbladder cancer. Seventeen proteins were successfully identified by MS, in which nine proteins were overexpressed in tumors and the other eight proteins were underexpressed. The positive expression rates of annexinA3 in gallbladder cancer was significantly higher than that in chronic cholecystitis, and the difference was statistically significant （74. 0% vs 21. 1% , P 〈 0. 01）. In the gallbladder cancer, no correlation was obtained between annexinA3 and age, gender or histologicl type （P 〉 0.05）, but overexpression of annexinA3 correlated significantly with those cases with a lower histological grading （40. 0% vs 82. 5% ,P 〈 0. 05）;lymph node or distant metastasis （40. 9% vs 100% , P 〈 0. 05） ;or a shorter survival time after operation （50. 0% vs 93. 8% , P 〈 0. 05）. Conclusions Significant discrepancies in protein expression exist among gallbladder cancer and benign gallbladder tissues. AnnexinA3 plays an important role in the initiation and progression of human gallbladder cancer.