用原位杂交及免疫组化方法检测p53基因及ki-67蛋白在胆管癌中的表达

Result of p53,ki-67 Protein Expression in Cholangiocarcinoma with in situ Hybridization and Immunohistochemistry Methods

目的 研究胆管癌组织中 p5 3基因表达、ki- 6 7增殖指数高低及它们之间的关系 ,以深入探讨胆管癌发生、发展的机制。方法 采用原位杂交及免疫组化方法对 30例人胆管癌组织 p5 3基因、P5 3蛋白表达及 ki- 6 7蛋白增殖指数进行检测 ,探讨它们之间的关系 ,并进而分析它们与临床病理资料之间的相关性。结果 原位杂交检测30例胆管癌组织中 14例 (46 .7% ) p5 3基因表达阳性 ,p5 3原位杂交结果与胆管癌的临床病理资料无显著相关性。免疫组化检测 30例胆管癌组织中 16例 (5 3.3% )有 P5 3蛋白表达 ,有局部淋巴结转移组 P5 3蛋白表达阳性率明显高于无局部淋巴结转移组 ;临床 期患者 P5 3蛋白表达阳性率明显高于临床 、 期患者。有 P5 3蛋白表达的患者生存期明显短于无 P5 3蛋白表达的患者。 2 9例 (96 .7% )胆管癌患者的肿瘤细胞中有 ki- 6 7蛋白表达。在中、低分化、有局部淋巴结转移、临床 期肿瘤组织中 ,ki- 6 7增殖指数显著高于高分化、无局部淋巴结转移及临床 、 期患者。生存期长的肿瘤患者 ,ki- 6 7增殖指数低于生存期短的患者。结论 ki- 6 7增殖指数反映了胆管癌细胞的增殖活性 ,分化程度低的肿瘤细胞增殖活跃 ,局部淋巴结转移的危险性较大 ,临床预后较差 ,提示 ki- 6

Objective To clarify the clinical significance of P53,ki 67,and the relationship between p53 gene expression and ki 67 labeling index. Methods In this study,30 cases of human cholangiocarcinoma,paired with normal bile duct tissue adjacent to the tumor,were included. In situ hybridization (ISH) was used to detect the p53 gene expression.Immunohistochemistry (IHC) was applied to analyze p53 gene mutation and ki 67 labelling index. Results ki 67 and P53 positivity were detected in 96 7% (29/30) and 53 3% (16/30) of carcinoma specimens respectively by IHC,neither in peripheral normal tissue.p53 positivity was related to local lymph node metastasis.p53 positivity of cholangiocarcinoma in clinical stage Ⅲ was significantly higher than that in clinical stage Ⅰ,Ⅱ.The expression of ki 67 protein was correlated with local lymph node metastasis and clinical stage.In lymph node metastasis group and clinical stage Ⅲ group,ki 67 labeling index was significantly higher than that in non lymph node metastasis group as well as in early stage group.We also found ki 67 labeling index of moderately or poorly differentiated cholangiocarcinoma was significantly higher than that of well differentiated carcinoma.Comparing survival time,the median survival time of patients whose bile duct tumors expressed no P53 protein or with lower ki 67 index was longer than that of patients whose tumors expressed P53 protein, or with higher ki 67 labeling index. Conclusions The results indicate that p53 positivity may be correlated with tumor development,growing activity.They also indicate that ki 67 labeling index reflects the degree of tumor cell proliferation.This observation suggests that ki 67 labeling index may be a useful predictor of tumor prognosis.