实时PCR检测卵巢癌石蜡块组织中Her-2/neu的新方法

Real-time PCR—an alternative method for Her-2/neu analysis in paraffin-embedded ovarian carcinoma

目的探索检测卵巢癌石蜡块组织中Her-2/neu状态的新方法。方法选取69例卵巢癌组织蜡块,用实时PCR检验卵巢癌组织中的Her-2/neu基因的扩增情况,用免疫组化检测其蛋白表达情况,将两者结果进行比较,并用FISH试验来验证其结果。结果肿瘤组织中Her-2/neu的扩增如果以相对基因拷贝数(肿瘤细胞内基因拷贝数与正常二倍体卵巢组织细胞基因拷贝数的比率)≥1.5或≥2为截取值,本组卵巢癌组织中Her-2/neu的扩增率分别为26.1%(18/69)、15.9%(11/69);本组卵巢癌组织中Her-2/neu免疫组化为++^+++者为28.99%(20/69);两种检测方法的结果具有很高的一致性。结论实时PCR简便、客观、高效,可望成为卵巢癌石蜡组织中除免疫组化外检测Her-2/neu变化的一种备选方法。

Objective To explore an alternative method for Her-2/neu analysis in paraffin-embedded ovarian carcinoma. Methods The amplification and expression of Her-2/neu in 69 specimens of ovarian carcinoma was investigated by real-time PCR （RT-PCR） and immunohistochemistry （IHC）, respectively. The results were compared and FISH assay was performed to ensure the reliability of the results. Results Based on arbitrary cut-offs of more than 1.5 or 2 （ratio of gene copy numbers in tumor cells to gene copy numbers in euploid cells）, the amplification rates for the Her-2/neu were 26.1% （18/69） and 15.9% （11/69） by RT- PCR. Twenty out of 69 specimens （28.99%） of Her-2/neu overexpression （ ＋＋ to ＋＋＋ ） were found by IHC. Her-2/neu status assessed by RT-PCR was closely in accordance with the result by IHC. Conclusion RT- PCR is reliable, easy and rapid to perform and represents an alternative technique for establishing Her-2/neu status in paraffin-embedded ovarian carcinoma