免疫组化与原位杂交双标记技术在鼻咽癌诊断中的应用

Application of Double Labeling of Immunohistochemistry Staining and in Situ Hybridization in Diagnosis of Nasopharyngeal Carcinoma

目的探讨如何应用免疫组化和原位杂交双重标记技术,对辅助鼻咽癌的组织学诊断最有帮助。方法采用免疫组化和原位杂交双重标记技术检测30例鼻咽未分化型非角化性癌组织,并在同一切片上检测细胞角蛋白(CK)和EB病毒(EBER),同时观察两种指标在鼻咽癌细胞中的表达情况。结果 CK定位于鼻咽癌细胞胞浆内,呈棕黄色;EBER阳性反应定位于鼻咽癌细胞的细胞核上,呈蓝黑色。30例鼻咽未分化型非角化性癌癌组织均呈CK和EBER阳性,可看到鼻咽癌细胞同时表达CK和EBER,CK的棕黄色和EBER的蓝黑色对比清晰容易观察。增生的鼻咽上皮细胞CK阳性,而EBER呈阴性。结论 CK免疫组化与EBER原位杂交双标记技术有助于鼻咽癌的组织学诊断,可常规作为鼻咽癌病理诊断常用的辅助技术。双标技术以先做EBER原位杂交,后做CK免疫组化,显色剂分别用BCIP/NBT和DAB为好。

Objective To investigate application of double labeling of im.munohistochemistry staining and in situ hybridization in improving histological diagnosis of nasopharyngeal carcinoma （NPC）. Methods Both cytokeratin （CK） and Epstein-Ban vires-encoded small RNAs （EBER） expression were detected on same NPC sections from 30 samples of undifferentiated type non-keratinizing NPC by double labeling ofimmunohistochemistry staining and in situ hybridization. Results CK positive signals, which present brown-yellow color, were located in cytoplasm of NPC cells; EBER positive signals, which showed blue-black, were located in nuclei of NPC cells. All 30 cases of NPC were positive for CK and EBER staining. Furthermore, both CK and EBER positive signals were found in same NPC cell. The two different positive signals showed remarkable different colors to identify easily. However, hyperplastic nasopharyngeal epithelium was positive for CK, negative for EBER. Conclusions Double labeling of immunohistochemistry staining and in situ hybridization is helpful for histological diagnosis of NPC and can be used in common. EBER expression is first detected by in situ hybridization, CK expression is then detected by immunohistochemistry staining in double labeling. BCIP/NBT and DAB is used as color developing agent, respectively.