Table1_MicroRNAs Regulating Tumor Immune Response in the Prediction of the Outcome in Patients With Breast Cancer.DOCX

MicroRNAs (miRNAs) are key regulators in immune surveillance and immune escape as well as modulators in the metastatic process of breast cancer cells. We evaluated the differential expression of plasma miR-10b, miR-19a, miR-20a, miR-126 and miR-155, which regulate immune response in breast cancer progression and we investigated their clinical relevance in the outcomes of breast cancer patients. Plasma samples were obtained from early (eBC; n = 140) and metastatic (mBC; n = 64) breast cancer patients before adjuvant or first-line chemotherapy, respectively. Plasma miRNA expression levels were assessed by qRT-PCR. We revealed a 4-miRNA panel consisted of miR-19a, miR-20a, miR-126, and miR-155 able to discriminate eBC from mBC patients with an AUC of 0.802 (p < 0.001). Survival analysis in eBC patients revealed that low miR-10b and miR-155 expression was associated with shorter disease free survival (disease free survival; p = 0.012 and p = 0.04, respectively) compared to high expression. Furthermore, miR-126 expression was associated with shorter overall survival (overall survival; p = 0.045). In multivariate analysis the number of infiltrated axillary lymph nodes and low miR-10b expression independently predicted for shorter DFS (HR: 2.538; p = 0.002 and HR: 1.943; p = 0.033, respectively) and axillary lymph nodes and low miR-126 for shorter OS (HR: 3.537; p = 0.001 and HR: 2.558; p = 0.018). In the subgroup of triple negative breast cancer (TNBC) patients, low miR-155 expression independently predicted for shorter DFS (HR: 5.056; p = 0.037). Accordingly in mBC, patients with low miR-10b expression had shorter progression free survival and OS compared to patients with high expression (p = 0.0017 and p = 0.042, respectively). In multivariate analysis, recurrent disease and low miR-10b expression independently predicted for shorter PFS (HR: 2.657; p = 0.001 and HR: 1.920; p = 0.017, respectively), whereas performance status two independently predicted for shorter OS (HR: 2.031; p = 0.03). In summary, deregulated expression of circulating miRNAs involved in tumor and immune cell interactions evaluated before adjuvant and 1st-line chemotherapy can distinguish disease status and emerge as independent predictors for outcomes of breast cancer patients.

微小RNA（microRNA，miRNA）是免疫监视与免疫逃逸的关键调控因子，同时也是乳腺癌细胞转移进程的调节因子。本研究评估了血浆miR-10b、miR-19a、miR-20a、miR-126及miR-155的差异表达水平——这些miRNA在乳腺癌进展中参与调控免疫应答，并探究了它们与乳腺癌患者临床结局的相关性。血浆样本分别取自辅助化疗前的早期乳腺癌（early breast cancer, eBC）患者（n=140）及一线化疗前的转移性乳腺癌（metastatic breast cancer, mBC）患者（n=64）。采用实时荧光定量逆转录聚合酶链反应（qRT-PCR）检测血浆miRNA表达水平。本研究构建了由miR-19a、miR-20a、miR-126及miR-155组成的四miRNA标志物组合，该组合可区分eBC与mBC患者，曲线下面积（AUC）为0.802（p<0.001）。对eBC患者的生存分析显示，与高表达组相比，miR-10b与miR-155低表达组的无病生存期（disease free survival, DFS）更短（分别为p=0.012与p=0.04）。此外，miR-126低表达与更短的总生存期（overall survival, OS）相关（p=0.045）。多因素分析显示，腋窝淋巴结浸润数目与miR-10b低表达可独立预测更短的DFS（风险比HR=2.538，p=0.002；HR=1.943，p=0.033）；而腋窝淋巴结浸润与miR-126低表达可独立预测更短的OS（HR=3.537，p=0.001；HR=2.558，p=0.018）。在三阴性乳腺癌（triple negative breast cancer, TNBC）患者亚组中，miR-155低表达可独立预测更短的DFS（HR=5.056，p=0.037）。同样，在mBC患者中，miR-10b低表达组的无进展生存期（progression free survival, PFS）与OS均短于高表达组（分别为p=0.0017与p=0.042）。在mBC的多因素分析中，复发疾病与miR-10b低表达可独立预测更短的PFS（HR=2.657，p=0.001；HR=1.920，p=0.017）；而体能状态评分为2分可独立预测更短的OS（HR=2.031，p=0.03）。综上，在辅助化疗与一线化疗前检测的、参与肿瘤与免疫细胞互作的循环miRNA表达失调，可区分疾病状态，并可作为乳腺癌患者临床结局的独立预测因子。