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运用实时荧光定量RT-PCR法检测乳腺癌组织中ERα、PR和Her2 mRNA表达
陆慧琦1,何金2,陈佳1,徐毅2,祝丽双1,韩焕兴1*
0
(1. 第二军医大学长征医院转化医学中心,上海 200003
2. 第二军医大学长征医院病理科,上海 200003
*通信作者)
摘要:
目的 评价实时荧光定量RT-PCR(qRT-PCR)在测定乳腺癌组织中雌激素受体α(ERα)、孕激素受体(PR)和人表皮生长因子受体2(Her2)基因表达的临床价值。方法 选择2010年3月至10月在我院接受手术治疗的乳腺癌患者48例和乳腺良性病变患者28例,用免疫组化染色(IHC)法检测乳腺癌组织中ERα、PR、Her2蛋白的表达,用实时qRT-PCR法检测乳腺癌组织及良性乳腺肿瘤组织中ERαPRHer2 mRNA的表达,并评价这两种检测方法在乳腺癌诊断中的价值。结果 用实时qRT-PCR 测得的ERα、Her2 mRNA在乳腺癌组织中的表达均高于对照组织(P<0.05,P<0.01),而PR mRNA的表达在乳腺癌组织与对照组织之间差异无统计学意义。乳腺癌组织中ERα、PR、Her2的蛋白表达与临床TNM分期有关,ERα和PR的阳性表达率随临床TNM分期的升高而下降(P<0.05),Her2的阳性表达率随临床TNM分期的升高而升高(P<0.05)。乳腺癌淋巴结转移组乳腺组织中ERαHer2 mRNA表达均高于无淋巴结转移组(P<0.05),而PR mRNA表达与无淋巴结转移组比较差异无统计学意义。实时qRT-PCR检测 ERαPRHer2 mRNA在评价乳腺癌对内分泌治疗的敏感性、特异性以及病理学诊断的符合率均与IHC相符。结论 ERα、PRHer2是预测乳腺癌内分泌治疗或靶向治疗的重要基因,本研究建立的实时qRT-PCR检测方法可用于ERα、PRHer2 mRNA表达的临床检测和研究。
关键词:  乳腺肿瘤  雌激素受体α  孕激素受体  人表皮生长因子受体2  实时荧光定量RT-PCR  预后
DOI:10.3724/SP.J.1008.2013.00453
投稿时间:2012-11-18修订日期:2013-01-11
基金项目:上海市科委基金(08411962100).
Real-time fluorescent quantitative reverse transcription polymerase chain reaction in detecting expression of ERα, PR and Her2 mRNA in breast cancer tissues
LU Hui-qi1,HE Jin2,CHEN Jia1,XU Yi2,ZHU Li-shuang1,HAN Huan-xing1*
(1. Center for Translational Medicine, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China
2. Department of Pathology, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China
*Corresponding author.)
Abstract:
Objective To evaluate the clinical value of real-time fluorescent quantitative reverse transcription polymerase chain reaction (qRT-PCR) in detecting expression of estrogen receptor alpha (ERα), progesterone receptor (PR) and human epidermal growth factor receptor 2 (Her2) genes in breast cancer tissues. Methods Totally 48 breast cancer tissues and 28 benign breast tumor tissues (control) were obtained from patients undergoing surgery in our hospital during Mar. 2010 and Oct. 2010. The expression of ERα, PR and Her2 protein was examined by immunohistochemistry (IHC) in breast cancer tissues and the expression levels of ERα, PR and Her2 mRNA were detected by real-time qRT-PCR in breast cancer tissues and benign breast tumor tissues. The values of these two methods in diagnosis of breast cancer were evaluated.Results The expressions of ERα and Her2 mRNA were significantly higher in the breast cancer tissues than in the controls (P<0.05, P<0.01), while the expression of PR mRNA had no significant difference between controls and breast cancer tissues. ERα, PR and Her2 protein expressions were associated with the TNM stage of breast cancer, with the former two being negatively associated with TNM stages (P<0.05) and Her2 being positively associated with TNM stages (P<0.05). The expressions of ERα and Her2 mRNA were significantly higher in breast cancer tissues of patients with lymph node metastasis than in those without lymph node metastasis (P<0.05). The expression of PR mRNA was not significantly different in breast cancer tissues between patients with and without lymph node metastasis (P>0.05). Real-time qRT-PCR in detecting ERα, PR and Her2 mRNA expression had similar capability with IHC method in evaluating the sensitivity, specificity of endocrine therapy; moreover, the two methods also had a consistent pathological diagnosis rates (P>0.05). Conclusion ERα, PR and Her2 genes are important predictive markers for endocrine therapy or targeted therapy of breast cancer. Real-time qRT-PCR method can be used for clinical detection and research of ERα, PR and Her2 mRNA.
Key words:  breast neoplasms  estrogen receptor alpha  progesterone receptor  human epidermal growth factor receptor 2  real-time fluorescent quantitative RT-PCR  prognosis