乳腺癌ER功能状态及腋淋巴结转移的检测

Assay of functional ER and breast cancer micrometastasis in axillary lymph nodes

目的 探讨ER功能检测及RT PCR检测乳癌淋巴结转移的临床应用。方法 应用凝胶迁移阻滞法及配体亲和组织化学法检测乳癌患者ER的功能状态。以患者腋窝淋巴结的RNA作模板 ,应用特异性引物扩增相应的Keratin19片段 ,据此判断有无腋窝淋巴结转移。结果 在 5 5例ER阳性的标本中 ,ERE结合阳性者达 71%。在HE染色淋巴结阴性的患者中 ,有 12 .8%经RT PCR检测阳性。结论 ERE结合功能有助于全面反映患者ER功能状态。RT PCR检测淋巴结Keratin19表达可作为乳腺癌转移的标志。

Objective This study is to evaluate the assay of functional ER by DNA binding analysis and clinical application of RT-PCR for detection of breast cancer micrometastasis to axillary lymph nodes.Methods Protein and total RNA extracted from primary breast cancer and axillary lymph nodes was subjected to perform the gel shift binding analysis and RT-PCT amplifying the Keratin19 fragments respectively. The functional condition of ER was estimated by gel shift binding analysis results combined with hormone binding histochemical assay. The lymph nodes metastasis was assessed referring to the results of both RT-PCR and hematoxylin-eosin (HE) staining. All patients were monitored for tumor relapse during the 3-year follow-up period.Results Among 55 ER positive breast cancer patients. 71% was ERE binding analysis positive. Keratin19 were detected in all 86 breast cancer specimens but not in lymph nodes from patients with benign diseases. Of patients with HE negative staining of lymph nodes, 12.8% (11/86) had significant Keratin19 expression.Conclusion The DNA binding analysis for ER by gel shift can be used as functional ER assay combined with hormone binding histochemical analysis. The expression of Keratin19 in lymph nodes detected by RT-PCR can be regarded as a sign of breast cancer micrometastasis, which conduced to determine the postoperative treatment and predict the prognosis for patients.