Zhu Chengjun, Xu Jing, Sun Jinyu, Cui Shiyun, Sun Yue, Yu Tao, Wang Cenzhu, Wang Tianyao, Wu Yufeng, Ju Feng, Yao Jiafeng, Liu Kai, Zhang Wenwen, Guan Xiaoxiang
Department of Oncology, The First Affiliated Hospital of Nanjing Medical University, No. 300 Guangzhou Road, Nanjing 210029, China.
Stomatological College, Nanjing Medical University, Nanjing 210029, China.
J Clin Med. 2022 Oct 14;11(20):6067. doi: 10.3390/jcm11206067.
Aims: This study investigates the association between circulating tumor cells (CTCs) and breast cancer metastasis. Methods: A retrospective study was conducted using patients with histologically confirmed breast cancer recruited from the First Affiliated Hospital of Nanjing Medical University during the period of August 2017−October 2020. We used adjusted logistic regression, the random forest algorithm, and sensitivity analysis to study the association between CTC enumeration and tumor metastasis. Further, we performed next-generation sequencing (NGS) on the CTCs obtained from two patients with breast cancer brain metastasis. Results: A total of 41 out of 116 enrolled patients were identified with tumor metastasis. CTC enumeration was significantly higher in patients with liver metastasis than in those without liver metastasis. Patients with CTCs ≥ 5 exhibited a higher risk of tumor metastasis than those with CTCs < 5 in the adjusted model (odds ratios (OR) = 6.25, 95% confidence interval (CI) = 2.63−15.58). The random forest model identified CTC enumeration as a significant metastasis-related variable with the highest mean decrease accuracy and mean decrease Gini score. No significant association was found between CTCs and visceral metastasis with an OR of 1.29 (95% CI = 0.98−2.05, p = 0.232). Upon further investigating organ-specific metastasis, we found that patients with high CTC levels were more likely to develop liver metastasis (OR = 4.87, 95% CI = 1.34−20.17, p = 0.021). The NGS study of CTCs identified a total of 120 indel mutations (e.g., CNGB1, NTSR1, ZG16). The enriched biological processes were mechanoreceptor differentiation and macrophage activation involved in the immune response. The enriched KEGG pathways included focal adhesion, the PI3K-Akt signaling pathway, and microRNAs involved in cancer. Conclusions: Our study revealed that CTCs ≥ 5 are a risk factor for tumor metastasis in breast cancer patients. In addition, we reported that CTCs ≥ 5 might be associated with a higher risk of liver metastasis in patients with metastatic breast cancer. We have provided the mutational profiles of CTCs based on next-generation sequencing.
本研究调查循环肿瘤细胞（CTC）与乳腺癌转移之间的关联。方法：进行一项回顾性研究，使用2017年8月至2020年10月期间从南京医科大学第一附属医院招募的组织学确诊乳腺癌患者。我们使用调整后的逻辑回归、随机森林算法和敏感性分析来研究CTC计数与肿瘤转移之间的关联。此外，我们对两名乳腺癌脑转移患者的CTC进行了二代测序（NGS）。结果：116名入组患者中共有41名被确定有肿瘤转移。肝转移患者的CTC计数显著高于无肝转移患者。在调整模型中，CTC≥5的患者比CTC<5的患者发生肿瘤转移的风险更高（优势比（OR）=6.25，95%置信区间（CI）=2.63 - 15.58）。随机森林模型将CTC计数确定为一个与转移相关的重要变量，其平均降低准确率和平均降低基尼系数最高。未发现CTC与内脏转移之间存在显著关联，OR为1.29（95%CI = 0.98 - 2.05，p = 0.232）。在进一步研究器官特异性转移时，我们发现CTC水平高的患者更有可能发生肝转移（OR = 4.87，95%CI = 1.34 - 20.17，p = 0.021）。对CTC的NGS研究共鉴定出120个插入缺失突变（如CNGB1、NTSR1、ZG16）。富集的生物学过程是参与免疫反应的机械感受器分化和巨噬细胞活化。富集的KEGG通路包括粘着斑、PI3K - Akt信号通路以及参与癌症的微小RNA。结论：我们的研究表明，CTC≥5是乳腺癌患者肿瘤转移的一个危险因素。此外，我们报告称，CTC≥5可能与转移性乳腺癌患者发生肝转移的较高风险相关。我们基于二代测序提供了CTC的突变谱。
