| 摘要: |
| 目的：应用显色原位杂交技术(chromogenic in situ hybridization,CISH)检测乳腺癌组织人表皮生长因子受体2(human epidermal growth factor receptor 2, HER2) 基因扩增情况，并对操作方法进行优化。方法：选择免疫组织化学（IHC）EnVision法检测结果2+及以上的60例乳腺癌组织作为研究对象，应用CISH技术检测其HER2基因扩增情况，分析二者结果的相关性；总结CISH操作经验，优化操作方法。结果：44例标本IHC检测HER2蛋白表达为3+，其中40例CISH检测呈现HER2基因高倍扩增，4例无扩增；基因扩增与蛋白表达的符合率为91％。16例IHC检测HER2表达为2+标本基因扩增与蛋白表达的符合率为50％。IHC与CISH符合率为80%(48/60)，二者明显相关(P＜0.01)。操作过程中应严格控制切片厚度，一般要求4～5 μm；杂交变性一定要彻底，杂交后的洗涤温度和时间也很重要，最好控制在70~75℃；要严格控制苏木精衬染时间；每次实验最好有阳性对照片，可以有效进行质量控制。结论：CISH检测乳腺癌HER2基因扩增的结果与IHC检测的蛋白表达结果符合率较高，可作为乳腺癌HER2基因检测的一项新技术；但在具体操作时，应注意严格控制切片厚度、消化及杂交后洗涤温度与时间以及苏木精衬染时间等因素。 |
| 关键词: 显色原位杂交技术 乳腺肿瘤 人表皮生长因子受体2 c-erbB-2基因 |
| DOI：10.3724/SP.J.1008.2008.00099 |
| 投稿时间：2007-03-20修订日期：2008-01-07 |
| 基金项目: |
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| A method for detection of HER2 gene amplification in breast cancer tissues: chromogenic in situ hybridization method and its modification |
| NI Can-rong*, BAI Chen-guang |
| (Department of Pathology, Changhai Hospital, Second Military Medical University, Shanghai 200433, China) |
| Abstract: |
| Objective：To apply chromogenic in situ hybridization (CISH) for detection of HER2 gene amplification in breast cancer tissues and to discuss some modifications of the CISH method. Methods: HER2 gene amplification was detected by CISH in 60 breast cancer specimens with an immunohistochemical score over 2+. The correlation between the results of IHC and CISH was analyzed.Our experience in CISH manipulation was summarized and optimization to CISH was discussed. Results: CISH identified gene amplification in 91％(40/44) specimens with an IHC score of 3+ and in 50％(8/16) specimens with an IHC score of 2+．The total concordance rate between IHC and CISH was 80％(48/60, P<0.01)．The thickness of sections should be controlled within 4-5 μm; the denaturation should be complete; and the post-hybridization washing temperature and time were also very important and the temperature should be controlled at 70-75℃. The dyeing time of hematoxylin should also be restrictedly controlled. Positive control should be set up in the experiment for high quality of the experiment. Conclusion: CISH has high concordance rate with IHC in examining HER2 amplification and it may be a new method for detection of HER2 gene. The thickness of the sections, the post-hybridization washing temperature and time, and the time of hematoxylin dyeing should be strictly controlled. |
| Key words: chromogenic in situ hybridization breast neoplasms human epidermal growth factor receptor 2 c-erbB-2 gene |
