应用荧光原位杂交法检测乳腺癌石腊样本中HER.docVIP

　　应用荧光原位杂交法检测乳腺癌石腊样本中HER 【摘要】 目的 探讨荧光原位杂交法（Fluorescence in situ hybridization, FISH）检测乳腺癌HER2基因扩增在临床病理诊断及分子靶向 治疗 中 应用 的可能性。 方法 用FISH技术和免疫组化（Immunohistochemistry，IHC）技术检测50例乳腺导管癌石蜡包埋标本并比较两种方法的结果以及与临床病理的关系。结果 16/50例HER2蛋白表达阳性，其中强阳性5例，中度阳性9例，弱阳性2例；11/50（22%）例乳腺癌标本FISH技术检测HER2基因扩增阳性，其中5/5为免疫组化HER2蛋白强阳性病例；6/9为中度阳性病例，其中1例为17号染色体多倍体与HER2基因扩增。HER2基因扩增与蛋白表达与乳腺癌转移有关（Plt;0.05）。结论 FISH技术可稳定地检测用IHC确定的HER2蛋白阳性乳腺癌中HER2基因的扩增状况，并用于临床赫赛汀分子靶向治疗病例的筛选。 【关键词】 HER2/癌基因；荧光原位杂交；乳腺癌 　　Application of Fluorescent in Situ Hybridization (FISH) to Detect the Amplification of HER2 Gene in Paraffin Sample of Breast Cancer 　　Abstract：Objective To explore the possibility of application ethod to detect amplification of HER2 gene by fluorescence in situ hybridization (FISH) in clinical pathology and molecular targeting therapy in breast cancer, and investigate the relationship betplification and clinicopathological data.Methods Detecting 50 cases of formalinfixed and paraffinembeded breast ductal carcinoma samples pared munohistochemistry and then analyzed a presented positive for HER2 protEin expression, including 5 strong, 9 moderate and 2 plification, in oderately positive expression for HER2 protEIn by immunohistochemistry. And the one plification of HER2 gene and polysomy of chromosome 17 oderately positive expression for the protein. Both gene amplification and protein expression of HER2 etastasis of lymph nodes (Plt;0.05).Conclusion FISH technique can stably detect the amplification of HER2 gene especially for the positive cases for HER2 protein and it is clinically useful to select the cases for molecular targeting therapy of Herceptin in breast cancer. 　　Key 厚切片,二甲苯脱蜡，100%乙醇脱二甲苯后空气干燥或45～50℃烤片2～5min。0.2N HCL 20min，经纯水充分洗涤后，2×SSC 缓冲液洗片3min；1M NaS（异硫酸氰钠） 80℃ 30min（温度要求准确）,纯水及缓冲液洗片后，将组织片放入蛋白酶溶液 37℃ 10min（250mg/500ml 0.01NHCL）。纯水洗涤后,切片入中性福尔马林10min。2×SSC缓冲液洗，45℃～50℃ 2～5min干燥切片；处理后的切片进入72℃变性液5min；经75%、85%、100%酒精各1min，空气中干片；滴加10μl探针混合液在组织片靶区域，上盖18×18mm盖玻片并用蜡胶封边。放入原位杂交仪中,37℃过夜（14～