Supplementary Material for: Increased Expression of Cyclooxygenase-2 and Increased Infiltration of Regulatory T Cells in Tumors of Patients with Hepatocellular Carcinoma

Background: Studies on Cox-2 and Foxp3+ regulatory T cells (Treg) in hepatocellular carcinoma (HCC) showed that Treg suppress local immune response in a Cox-2-dependent fashion. Aims: To investigate Cox-2 expression, Foxp3+ Treg infiltration and CD4+ T cell frequency in HCC tumors. Methods: Tumors and the corresponding nontumor hepatitis B virus-related liver tissues from 40 HCC patients with hepatitis B virus infection, plus 10 liver samples from patients with hemangioma as controls, were assessed for Cox-2 expression, Foxp3+ Treg and total CD4+ T cell numbers using immunohistochemistry. Serum TGF-β1 was assessed by ELISA. Results: Reduced Cox-2 expression, increased Treg and increased CD4+ T cells were shown in tumor as compared with nontumor tissues. Moreover, of 40 tumor tissues, 23 that expressed Cox-2 showed increased Foxp3+ Treg and reduced CD4+ T cells compared with the remaining 17 that did not express Cox-2. Correlation analyses showed that within tumors Treg infiltration correlated positively with Cox-2 expression, and that Treg infiltration or Cox-2 expression correlated negatively with CD4+ T cells. Additionally, serum TGF-β1 was higher in HCC patients than in controls. Conclusion: Within tumors, Cox-2 expression, Treg infiltration and CD4+ T cell frequency were increased, and the Cox-2 expression correlated positively with Treg infiltration and negatively with CD4+ T cell frequency.

背景：针对肝细胞癌（hepatocellular carcinoma, HCC）中环氧合酶-2（Cyclooxygenase-2, Cox-2）与Foxp3+调节性T细胞（Foxp3+ regulatory T cells, Treg）的相关研究表明，Treg可通过Cox-2依赖的途径抑制局部免疫应答。研究目的：探讨肝细胞癌肿瘤组织中的Cox-2表达水平、Foxp3+ Treg浸润情况及CD4+T细胞占比。方法：纳入40例合并乙型肝炎病毒感染的肝细胞癌患者的肿瘤组织及配对的非肿瘤性乙型肝炎病毒相关肝组织，另以10例肝血管瘤患者的肝组织作为对照。采用免疫组化（immunohistochemistry）法检测各组样本的Cox-2表达水平、Foxp3+ Treg及总CD4+T细胞数量；采用酶联免疫吸附试验（enzyme-linked immunosorbent assay, ELISA）检测血清转化生长因子-β1（TGF-β1）水平。结果：与非肿瘤组织相比，肝细胞癌肿瘤组织的Cox-2表达水平降低，而Foxp3+ Treg及CD4+T细胞数量均升高。在40例肿瘤组织中，23例Cox-2阳性表达的样本，其Foxp3+ Treg浸润程度高于其余17例Cox-2阴性表达的样本，而CD4+T细胞数量则更低。相关性分析结果显示，肿瘤组织内的Foxp3+ Treg浸润程度与Cox-2表达水平呈正相关，Foxp3+ Treg浸润程度或Cox-2表达水平均与CD4+T细胞数量呈负相关。此外，肝细胞癌患者的血清TGF-β1水平显著高于对照组。结论：在肝细胞癌肿瘤组织内，Cox-2表达、Foxp3+ Treg浸润程度及CD4+T细胞占比均存在异常变化，且Cox-2表达水平与Foxp3+ Treg浸润程度呈正相关，与CD4+T细胞占比呈负相关。