Shen H, Sheng H, Lu J J, Feng C, Yao M, Pan H, Xu L S, Shen J F, Zheng Y, Zhou Y L
Central Laboratory, the First Hospital of Jiaxing, Jiaxing 314001, China.
Zhonghua Yi Xue Za Zhi. 2018 May 8;98(17):1352-1357. doi: 10.3760/cma.j.issn.0376-2491.2018.17.014.
To explore the expression and distribution of programmed death receptor 1 (PD-1) and T-cell immunoglobulin mucin 3 (TIM-3) in breast cancer microenvironment and analyze the their correlation with the clinicopathological features. The specimens of tumor tissue and adjacent tissues from 30 patients with infiltrative breast cancer who were diagnosed as breast cancer from June 2016 to May 2017 in The First Hospital of Jiaxing were collected, and the specimen were divided into two parts along the center. After embedding and cryosectioning, the expression and distribution of PD-1 and TIM-3 protein in tumor tissues were observed by immunofluorescence staining. Another part of the specimen was cut and digested, and non-continuous density gradient centrifugation was used to extract tumor-infiltrating lymphocytes (TILs), real-time quantitative PCR (qRT-PCR) was used to detect the mRNA expression of PD-1 and TIM-3 in TILs. Meanwhile, the protein expression was determined by Western blotting. The relationship between the expression of PD-1 and TIM-3 and pathological parameters of breast cancer was analyzed with correlation analysis. Immunofluorescence results showed that more PD-1 and TIM-3 positive cells were observed in the tumor tissues compared with the tumor-adjacent tissues. The qRT-PCR showed that the expression of PD-1 and TIM-3 mRNA in TILs were both significantly higher than those in paracancerous tissues (3.09±0.38 vs 1.26±0.23, 3.42±0.31 vs 1.57±0.29, =4.16, 4.37, both <0.05). At the protein level, the expression of PD-1 and TIM-3 in tumor tissue lymphocytes(0.66±0.08, 0.80±0.11) was significantly higher than those in cancerous tissues(0.10±0.01, 0.26±0.02) (=6.79, 4.57, both <0.05). There were significant differences in the expression of PD-1, TIM-3 mRNA in the TILs between the different tumor histological grades, tumor sizes, lymph node metastasis (=2.22-2.99, all <0.05). Correlation analysis showed that there was a significant positive correlation between the expression of PD-1 and TIM-3 in tumor tissues (=0.616, <0.01). In the breast cancer microenvironment, PD-1, TIM-3-mediated signaling pathway plays an important role in the occurrence and development of breast cancer, it provides a new basis for the combination therapy of breast cancer.
探讨程序性死亡受体1（PD-1）和T细胞免疫球蛋白黏蛋白3（TIM-3）在乳腺癌微环境中的表达及分布，并分析其与临床病理特征的相关性。收集2016年6月至2017年5月在嘉兴市第一医院确诊为浸润性乳腺癌的30例患者的肿瘤组织及癌旁组织标本，将标本沿中心分为两部分。经过包埋和冰冻切片后，采用免疫荧光染色观察肿瘤组织中PD-1和TIM-3蛋白的表达及分布。将另一部分标本进行切割消化，采用不连续密度梯度离心法提取肿瘤浸润淋巴细胞（TILs），采用实时定量PCR（qRT-PCR）检测TILs中PD-1和TIM-3的mRNA表达。同时，采用蛋白质印迹法测定蛋白表达。采用相关性分析分析PD-1和TIM-3的表达与乳腺癌病理参数之间的关系。免疫荧光结果显示，与癌旁组织相比，肿瘤组织中观察到更多的PD-1和TIM-3阳性细胞。qRT-PCR结果显示，TILs中PD-1和TIM-3 mRNA的表达均显著高于癌旁组织（3.09±0.38 vs 1.26±0.23，3.42±0.31 vs 1.57±0.29，P=4.16，4.37，均<0.05）。在蛋白水平，肿瘤组织淋巴细胞中PD-1和TIM-3的表达（0.66±0.08，0.80±0.11）显著高于癌旁组织（0.10±0.01，0.26±0.02）（P=6.79，4.57，均<0.05）。不同肿瘤组织学分级、肿瘤大小、淋巴结转移的TILs中PD-1、TIM-3 mRNA的表达存在显著差异（P=2.22~2.99，均<0.05）。相关性分析显示，肿瘤组织中PD-1和TIM-3的表达之间存在显著正相关（r=0.616，P<0.01）。在乳腺癌微环境中，PD-1、TIM-3介导的信号通路在乳腺癌的发生发展中起重要作用，为乳腺癌的联合治疗提供了新的依据。
