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一种用于定量3种生存素转录本的实时定量聚合酶链反应方法的验证及在乳腺癌组织中的评估

Validation of a real-time quantitative polymerase chain reaction method for the quantification of 3 survivin transcripts and evaluation in breast cancer tissues.

作者信息

Pavlidou Anastasia, Kroupis Christos, Goutas Nikolaos, Dalamaga Maria, Dimas Kleanthi

机构信息

Department of Clinical Biochemistry, University of Athens Medical School, Attikon University Hospital, Haidari, Greece.

Department of Clinical Biochemistry, University of Athens Medical School, Attikon University Hospital, Haidari, Greece.

出版信息

Clin Breast Cancer. 2014 Apr;14(2):122-31. doi: 10.1016/j.clbc.2013.10.012. Epub 2013 Oct 27.

Abstract

BACKGROUND

Survivin is a novel antiapoptotic gene, which is a member of the inhibitor of apoptosis protein (IAP) family. Recently, 3 splice variants of this gene were cloned and characterized. This study aimed to validate a sensitive and specific method for the detection of survivin variants in breast cancer.

METHODS

Real-time quantitative polymerase chain reaction (qPCR) was performed on the cDNA with a reverse primer specific for each splice variant and a pair of common hybridization probes.

RESULTS

The expression of wild-type survivin was significantly correlated with survivin-2b, survivin-ΔEx3, and the ratio of survivin-ΔEx3 to wild-type survivin (P < .001). The ratio of survivin-2b to wild-type survivin was strongly associated with the ratio of survivin-ΔEx3 to wild-type survivin (P < .001). There was a strong positive association between the grade of the tumor and survivin-2b mRNA, survivin-ΔEx3 mRNA, and the ratio of survivin-ΔEx3 to wild-type survivin mRNA (P < .05). The ratio of survivin-2b to wild-type survivin was significantly associated with the presence of estrogen receptors (P = .05).

CONCLUSION

Our validated data suggest that survivin isoforms may be related to clinicopathological features and could be used as molecular prognostic tools or as new therapy targets.

摘要

背景

生存素是一种新型抗凋亡基因,属于凋亡抑制蛋白(IAP)家族成员。最近,该基因的3种剪接变体被克隆并鉴定。本研究旨在验证一种检测乳腺癌中生存素变体的灵敏且特异的方法。

方法

使用针对每种剪接变体的反向引物和一对通用杂交探针,对cDNA进行实时定量聚合酶链反应(qPCR)。

结果

野生型生存素的表达与生存素-2b、生存素-ΔEx3以及生存素-ΔEx3与野生型生存素的比值显著相关(P <.001)。生存素-2b与野生型生存素的比值与生存素-ΔEx3与野生型生存素的比值密切相关(P <.001)。肿瘤分级与生存素-2b mRNA、生存素-ΔEx3 mRNA以及生存素-ΔEx3与野生型生存素mRNA的比值之间存在强正相关(P <.05)。生存素-2b与野生型生存素的比值与雌激素受体的存在显著相关(P =.05)。

结论

我们验证的数据表明,生存素异构体可能与临床病理特征相关,可作为分子预后工具或新的治疗靶点。

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