Yoshinami Tetsuhiro, Kagara Naofumi, Motooka Daisuke, Nakamura Shota, Miyake Tomohiro, Tanei Tomonori, Naoi Yasuto, Shimoda Masafumi, Shimazu Kenzo, Kim Seung Jin, Noguchi Shinzaburo
Department of Breast and Endocrine Surgery, Osaka University Graduate School of Medicine, 2-2-E10 Yamadaoka, Suita, Osaka 565-0871, Japan.
Department of Breast and Endocrine Surgery, Osaka University Graduate School of Medicine, 2-2-E10 Yamadaoka, Suita, Osaka 565-0871, Japan.
Transl Oncol. 2020 Aug;13(8):100787. doi: 10.1016/j.tranon.2020.100787. Epub 2020 May 27.
We attempted to detect circulating tumor DNA (ctDNA), taking advantage of molecular barcode next-generation sequencing (MB-NGS), which can be more easily customized to detect a variety of mutations with a high sensitivity than PCR-based methods. Sequencing with a gene panel consisting of the 13 most frequently mutated genes in breast tumors from stage I or II patients revealed 95 somatic mutations in the 12 genes in 62% (62/100) of tumors. Then, plasma DNA from each patient (n = 62) before surgery was analyzed via MB-NGS customized to each somatic mutation, resulting in the detection of ctDNA in 16.1% (10/62) of patients. ctDNA was significantly associated with biologically aggressive phenotypes, including large tumor size (P = .004), positive lymph node (P = .009), high histological grade (P < .001), negative ER (P = .018), negative PR (P = .017), and positive HER2 (P = .046). Furthermore, distant disease-free survival was significantly worse in patients with ctDNA (n = 10) than those without ctDNA (n = 52) (P < .001). Our results demonstrate that MB-NGS personalized to each mutation can detect ctDNA with a high sensitivity in early breast cancer patients at diagnosis, and it seems to have a potential to serve as a clinically useful tumor marker for predicting their prognosis.
我们尝试利用分子条形码下一代测序技术（MB-NGS）检测循环肿瘤DNA（ctDNA），相较于基于聚合酶链反应（PCR）的方法，MB-NGS能够更轻松地进行定制，以高灵敏度检测多种突变。对I期或II期乳腺癌患者中13个最常发生突变的基因组成的基因panel进行测序，结果显示，在62%（62/100）的肿瘤中，12个基因存在95个体细胞突变。然后，通过针对每个体细胞突变定制的MB-NGS对每位患者（n = 62）术前的血浆DNA进行分析，结果在16.1%（10/62）的患者中检测到了ctDNA。ctDNA与生物学侵袭性表型显著相关，包括肿瘤体积大（P = .004）、淋巴结阳性（P = .009）、组织学分级高（P < .001）、雌激素受体阴性（P = .018）、孕激素受体阴性（P = .017）以及人表皮生长因子受体2阳性（P = .046）。此外，ctDNA阳性患者（n = 10）的远处无病生存期显著短于ctDNA阴性患者（n = 52）（P < .001）。我们的结果表明，针对每个突变进行个性化定制的MB-NGS能够在早期乳腺癌患者诊断时高灵敏度地检测ctDNA，并且似乎有潜力作为预测其预后的临床有用肿瘤标志物。
