Bakaeean Behnaz, Gholamin Mehran, Tabatabaee Yazdi Seyed Abbas, Forghani Mohammad Naser
Department of Biology, Marvdasht Branch, Islamic Azad University, Marvdasht, Iran.
Department of Biology, Science and Research Branch, Islamic Azad University, Marvdasht, Iran.
Iran Biomed J. 2020 May;24(3):183-91. doi: 10.29252/ibj.24.3.183. Epub 2020 Jan 26.
Intra-operative molecular diagnostic assays are currently used for the detection of lymph node metastases. The objective of this study was to find new biomarkers to improve diagnostic accuracy in the detection of metastatic axillary lymph nodes in breast cancer patients.
We applied an absolute quantitative real-time reverse transcription-PCR to quantitate the expression of CK19, KLK11, and CLEC3A mRNAs in 79 FFPE sentinel lymph nodes (SLNs) from 35 breast cancer patients. The CK19 was confirmed as a standard biomarker, and the level of expression of selected new markers, KLK11 and CLEC3A, was evaluated in pathologically negative and positive SLNs by using absolute quantitative real-time PCR.
The overall concordance of the CK19 gene with pathological results was 92.4% (less than 250 copies) in negative SLNs and 85% in positive SLNs (more than 250 copies). The sensitivity and specificity of CK19, which were detected by real-time PCR, was 85% and 46%, respectively. Our results revealed that lower CLEC3A was associated with more lymph node involvement. We could set a cut-off point for CLEC3A with the sensitivity of 78% and specificity of 60%. Also, the mean KLK11 had a statistically significant reverse correlation with tumor grade (p = 0.017). Higher CK19 levels were related to more tumor invasion (p < 0.0001).
Regarding the findings, CLEC3A along with CK19 can be used as a promising marker with high sensitivity and specificity for the detection of metastatic SLN.
术中分子诊断检测目前用于检测淋巴结转移。本研究的目的是寻找新的生物标志物，以提高乳腺癌患者腋窝转移淋巴结检测的诊断准确性。
我们应用绝对定量实时逆转录聚合酶链反应（RT-PCR）来定量35例乳腺癌患者的79个福尔马林固定石蜡包埋（FFPE）前哨淋巴结（SLN）中CK19、KLK11和CLEC3A mRNA的表达。CK19被确认为标准生物标志物，并通过绝对定量实时PCR评估所选新标志物KLK11和CLEC3A在病理阴性和阳性SLN中的表达水平。
CK19基因与病理结果的总体一致性在阴性SLN中为92.4%（少于250拷贝），在阳性SLN中为85%（多于250拷贝）。实时PCR检测的CK19的敏感性和特异性分别为85%和46%。我们的结果显示，较低的CLEC3A与更多的淋巴结受累相关。我们可以为CLEC3A设定一个临界值，其敏感性为78%，特异性为60%。此外，KLK11的平均水平与肿瘤分级具有统计学显著的负相关（p = 0.017）。较高的CK19水平与更多的肿瘤侵袭相关（p < 0.0001）。
基于这些发现，CLEC3A与CK19一起可作为检测转移性SLN的具有高敏感性和特异性的有前景的标志物。
