Role of stromal collagen in immunomodulation and prognosis of advanced gastric carcinoma

Although several hypotheses have been proposed explaining the mechanisms of the immune-privileged status of malignant tumors, the exact pathway is yet to be explored. Tumor stroma plays a vital role in the prognosis of cancer patients; however, the immunomodulatory impact of gastric cancer stroma has not been reported. We have evaluated the amount of stromal collagen and its impact on the infiltration of immune-competent cells into the tumor cell nest in gastric carcinoma. Tissue specimens from 84 advanced gastric carcinoma patients who had undergone a curative resection were evaluated for host immune status (CD8+ T cells), tumor stromal reaction (AZAN staining), tumor Fas ligand expression and incidence of tumor cell apoptosis (by TUNEL). The number of apoptotic tumor cells (apoptotic index [AI]) increased proportionally with an increase in the number of CD8+ T cells within the cancer cell nest (nest CD8) (p = 0.0001). Nest CD8 was inversely correlated with the amount of stromal collagen (p < 0.0001). Nest CD8 and AI became independent predictors of patient survival (p = 0.0023 and p = 0.044, respectively) in Cox's multivariate analysis. The amount of stromal collagen was found to be a significant predictor of disease relapse in univariate analysis (p = 0.0010) but not in multivariate analysis (p = 0.4729). In conclusion, increased nest CD8 produced a survival advantage by inducing tumor cell apoptosis in gastric carcinoma patients. Increased tumor stromal collagen worked as a barrier for CD8+ T-cell infiltration and might be one of the mechanisms of tumor escape from the host immune attack.     

Prognostic value of intratumoral CD8+ T lymphocyte in extrahepatic bile duct carcinoma as essential immune response

BACKGROUND AND OBJECTIVES: Tumor-infiltrating lymphocytes form an important aspect of the host defense against an expanding neoplasm. CD8+ T cells have been identified as a prognostic factor in several cancers. Here, we investigate that the influence of CD8+ T cells on extrahepatic bile duct carcinoma (EBDC) patient survival. METHODS: CD8+ T cell immunoreactivity in 58 surgically resected EBDC specimens was investigated. The relationship between CD8+ T cell immunoreactivity and clinical and histopathologic features was analyzed. RESULTS: Thirty-two tumors (55%) possessed intratumoral CD8+ T cells. The degree of intratumoral CD8+ T cell immunoreactivity demonstrated a significant relationship to lower numbers of lymph node metastasis, reduced venous invasion, decreased perineural invasion, and better pTNM staging. Intratumoral CD8- T cells were also associated with increased patient survival. Multivariate analysis indicated that the presence of intratumoral CD8+ T cells was an independent prognostic factors. CONCLUSIONS: The infiltration of a cancer cell nest by CD8+ T cells is a reliable marker predicting increased survival of patients with EBDC.     

The immunological impact of preoperative chemoradiotherapy on the tumor microenvironment of pancreatic cancer

Several therapeutic regimens, including neoadjuvant chemoradiation therapy (NACRT), have been reported to serve as anticancer immune effectors. However, there remain insufficient data regarding the immune response after NACRT in pancreatic ductal adenocarcinoma (PDAC) patients. Data from 40 PDAC patients that underwent surgical resection after NACRT (NACRT group) and 30 PDAC patients that underwent upfront surgery (US group) were analyzed to examine alterations in immune cell counts/distribution using a multiplexed fluorescent immunohistochemistry system. All immune cells were more abundant in the cancer stroma than in the cancer cell nest regardless of preoperative therapy. Although the stromal counts of CD4+ T cells, CD20+ B cells, and Foxp3+ T cells in the NACRT group were drastically decreased in comparison with those of the US group, counts of these cell types in the cancer cell nest were not significantly different between the two groups. In contrast, CD204+ macrophage counts in the cancer stroma were similar between the NACRT and US groups, while those in the cancer cell nests were significantly reduced in the NACRT group. Following multivariate analysis, only a high CD204+ macrophage count in the cancer cell nest remained an independent predictor of shorter relapse-free survival (odds ratio = 2.37; P = .033). NACRT for PDAC decreased overall immune cell counts, but these changes were heterogeneous within the cancer cell nests and cancer stroma. The CD204+ macrophage count in the cancer cell nest is an independent predictor of early disease recurrence in PDAC patients after NACRT.     

Expression of coinhibitory receptors on T cells in the microenvironment of usual vulvar intraepithelial neoplasia is related to proinflammatory effector T cells and an increased recurrence‐free survival

Human papillomavirus‐induced usual‐type vulvar intraepithelial neoplasia (uVIN) are infiltrated by immune cells but apparently not cleared. A potential explanation for this is an impaired T cell effector function by an immunesuppressive milieu, coinfiltrating regulatory T cells or the expression of coinhibitory molecules. Here, the role of these potential inhibitory mechanisms was evaluated by a detailed immunohistochemical analysis of T cell infiltration in the context of FoxP3, Tbet, indoleamine 2,3‐dioxygenase, programmed cell death 1, T cell immunoglobulin mucin 3 (TIM3), natural killer cell lectin‐like receptor A (NKG2A) and galectins‐1, ?3 and ?9. Paraffin‐embedded tissues of primary uVIN lesions (n = 43), recurrent uVIN lesions (n = 20), vulvar carcinoma (n = 21) and healthy vulvar tissue (n = 26) were studied. We show that the vulva constitutes an area intensely surveyed by CD8+, CD4+, Tbet+ and regulatory T cell populations, parts of which express the examined coinhibitory molecules. In uVIN especially, the number of regulatory T cells and TIM3+ T cells increased. The expression of the coinhibitory markers TIM3 and NKG2A probably reflected a higher degree of T cell activation as a dense infiltration with stromal CD8+TIM3+ T cells and CD3+NKG2A+ T cells was related to the absence of recurrences and/or a prolonged recurrence‐free survival. A dense coinfiltrate with regulatory T cells was negatively associated with the time to recurrence, most dominantly when the stromal CD8+TIM3+ infiltration was limited. This notion was sustained in vulvar carcinoma's where the numbers of regulatory T cells progressively increased to outnumber coinfiltrating CD8+TIM3+ T cells and CD3+NKG2A+ T cells.     

Prognostic significance of CD8+ T cell and macrophage peritumoral infiltration in colorectal cancer

The purpose of this study was to evaluate the prognostic significance of CD8+ T cell and macrophage peritumoral infiltration in patients with colorectal cancer. A total of 97 adenocarcinomas of the colon and rectum were examined. Immunohistochemical staining was performed by the standard avidin-biotin-peroxidase complex method using antibodies to CD8 and CD68. Peritumoral infiltration by CD8+ T cells or macrophages was evaluated along the invasive margin of the cancer in each specimen. The area with the most abundant infiltration was selected, and the number of immunoreactive positive cells counted at x400 magnification. Patients were divided into two groups based on the degree of infiltration by each cell type: namely those with a high level of infiltration (more than the mean number of positive cells) and those with a low level of infiltration (less than the mean number of positive cells). Patients with a low level of macrophage infiltration had a significantly deeper depth of invasion than patients with a high level of macrophage infiltration (P=0.027). The percentage of patients with a high level of macrophage infiltration was significantly higher in vascular invasion-negative cases (46.7%) than in vascular invasion-positive cases (22.7%; P=0.045), and in lymph node metastasis-negative cases (52.9%) than in lymph node metastasis-positive cases (28.3%; P=0.014). Overall survival was significantly shorter for patients with a low level of CD8+ T cell infiltration than those with a high level of CD8+ T cell infiltration (P=0.01). The survival rate for patients with a high level of both CD8+ T cell and macrophage infiltration was 100%. In conclusion, both CD8+ T cell and macrophage peritumoral infiltration indicates anti-tumoral action in patients with colorectal cancer.     

Immunohistochemical study of leukocyte infiltration and expression of hsp70 in esophageal squamous cell carcinoma

It is reported that macrophages and CD4+ or CD8+ cytotoxic T cells have an important role in the suppression of cancer progression. The aim of this study was to clarify these immune responses in patients with esophageal cancer. We enrolled 28 patients with pT2 esophageal cancer that had been resected without preoperative adjuvant therapy. The correlations between the numbers of infiltrating CD4+, CD8+ and CD68+ cells, the expression of heat shock protein 70 (hsp70) and a variety of clinicopathologic factors were analyzed. The numbers of CD8+ T cells and CD68+ macrophages showed a significant positive correlation with tumor diameter (p = 0.01, p = 0.037) and the expression of hsp70 (p = 0.01, p = 0.02) and a negative correlation with lymph node metastasis (p = 0.0079, p < 0.0001). The expression of hsp70 exhibited a negative correlation with lymph node metastasis (p = 0.023). CD8+ T cells and CD68+ macrophages might have a suppressive function against esophageal cancer progression. Our results suggested that hsp70 might play an important role in the presentation of tumor specific antigens.     

Epstein-Barr virus LMP1 status in relation to apoptosis, p53 expression and leucocyte infiltration in nasopharyngeal carcinoma

BACKGROUND: Nasopharyngeal carcinoma (NPC) is consistently associated with Epstein-Barr virus (EBV) infection. EBV-encoded LMP1, expressed in most of NPC, has been suggested to have an important role in the pathogenesis and development of NPC and its expression correlates with poor prognosis. MATERIALS AND METHODS: Eighty-seven NPC biopsies were analyzed by immunohistochemistry for expression of markers of cell proliferation, apoptosis, infiltrating T lymphocytes and macrophages in relation to the LMP1 status. RESULTS: Our findings indicate that the p53 accumulation in NPC was significantly correlated to LMP1 and MMP9 overexpression in NPC cells. The frequency of apoptotic cells in NPC, as analyzed by TUNEL labeling, correlated to Fas-L and caspase-3 expression, and inversely to LMP1, p53 and MMP 9 expression. CD8+ T cell infiltration was predominately seen in nests of cancer cells with a high level of EBV-LMP1 expression, but these CD8+ T cells showed low expression of CD25 and TIA-1, indicating that they were not activated. CONCLUSION: Our observation suggests that the heavy infiltration by lymphocytes in LMP1-positive NPC tumors does not appear to counteract tumor growth by cytoxicity as indicated by the low apoptotic index. Thus, LMP1 seems to enhance survival- and proliferation-related signals in NPC. In analogy with other tumors, both the infiltrating T cells and the accumulated p53 may be inactive.     

Expression of cyclooxygenase-2 and inducible nitric oxide synthase correlates with tumor angiogenesis in endometrial carcinoma

The present study evaluated the significance of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) in 30 patients with endometrial carcinoma and the relationship of those molecular markers to tumor characteristics and microvessel density (MVD). Immunohistochemical expression of COX-2, iNOS, and CD34 was analyzed on paraffin-embedded tissue sections. The COX-2 and iNOS positive rates were 66.7% and 73.3%, respectively. The level of COX-2 expression was higher in grade II tumors than in grade III tumors (p < 0.05). The percentage of iNOS positivity was higher in patients with deep myometrial invasion than in patients without or less than 50% myometrial invasion (p < 0.05). There was significant correlation between positive COX-2 and positive iNOS expression (r = 0.601, p < 0.001). Both COX-2 and iNOS were significantly correlated with MVD (r = 0.02 p < 0.05; r = 0.599 p < 0.0001, respectively). The present findings suggest that combined expression of COX-2 and iNOS may play an important role in development and invasion of endometrial cancer and that this could be partially attributable to modulation of angiogenesis by COX-2 and iNOS.     

Coexpression of cyclooxygenase-2 and thymidine phosphorylase as a prognostic indicator in patients with FIGO stage IIB squamous cell carcinoma of uterine cervix treated with radiotherapy and concurrent chemotherapy

PURPOSE: To evaluate the prognostic significance of thymidine phosphorylase (TP) and coexpression of cyclooxygenase-2 (COX-2)/TP, and to investigate the relationship between COX-2 and TP expression in squamous cell carcinoma of the uterine cervix. METHODS AND MATERIALS: Cancer specimens from 75 patients with International Federation of Gynecology and Obstetrics Stage IIB squamous cell carcinoma of the uterine cervix who had undergone radiotherapy and concurrent chemotherapy were immunohistochemically stained with COX-2 and TP antibodies and scored. The prognostic significance of their expression status, and the relationship between COX-2 and TP was investigated. RESULTS: TP predominantly stained cytoplasm and the cell membrane of the tumor cells mainly in a diffuse and intense manner. TP was negative (<10% distribution) in 17%, 1+ (10-50%) in 25%, and 2+ (>50%) in 57% of patients. TP overexpression was related to a marginal prognostic significance of a poor 5-year overall survival (p = 0.082, log-rank test) and a high locoregional recurrence rate (p < 0.1, chi-square test). COX-2 and TP coexpression was observed in 24% of patients and was significantly related to poor 5-year disease-free and overall survival rates (p = 0.0083 and p = 0.025, respectively), a high pelvic lymph node involvement rate, a poor response to treatment, and a greater incidence of locoregional recurrence (p < 0.05). By multivariate analyses, only COX-2, TP, and coexpression of COX-2/TP were significant independent prognostic indicators of patient survival. All tumors showed 1+ or 2+ TP expression when COX-2 was positive, and no tumor expressed COX-2 when TP was negative (p = 0.03). In contrast, 77% of tumors expressed 1+ or 2+ TP without the synchronous expression of COX-2. CONCLUSIONS: Thymidine phosphorylase expression or COX-2/TP coexpression may be used as a molecular prognostic marker for squamous cell carcinoma of the uterine cervix. TP appears to be an important downstream molecule of COX-2 during angiogenesis and may be a new target for the treatment of uterine cervical cancer.     

基于生物信息学方法探讨Her-2基因表达对胃癌免疫细胞浸润的影响

目的 探讨人类表皮生长因子受体2（human epidermal growth factor receptor-2,Her-2）对胃癌组织中免疫细胞浸润的影响及可能的机制。方法 从癌症基因组图谱（the Cancer Genome Atlas,TCGA）数据库下载胃癌转录测序数据及临床资料。采用CIBERSORT反卷积算法获得胃癌微环境中浸润的免疫细胞占比,分析其与Her-2表达的关系,筛选Her-2基因潜在的调控浸润免疫细胞的类型。采用基因集变异分析（gene set variation analysis,GSVA）评估与胃癌组织Her-2表达相关的信号通路集合,利用GEO（gene expression omnibus）收录的胃癌转录组数据验证Her-2基因表达与胃癌免疫细胞浸润程度的关系。结果 CIBERSORT分析TCGA数据库胃癌样本发现CD8⁺T细胞、静息状态记忆性CD4⁺T细胞、M0型巨噬细胞及肥大细胞在Her-2高表达及低表达组中的浸润程度差异有统计学意义（P=0.029,0.049,0.027,0.008）。GEO数据库与TCGA数据库分析结果均显示,Her-2基因高表达与CD8⁺ T细胞、静息状态记忆性CD4⁺T细胞、M0型巨噬细胞及肥大细胞的发育、诱导分化、迁移调控有关（P<0.05）,并影响胃癌组织中部分免疫细胞的浸润能力和程度。结论 在胃癌中Her-2基因高表达可能对部分免疫细胞形成和功能发挥具有抑制作用。     

Cycloxygenase-2 inhibition augments the efficacy of a cancer vaccine.

Tumor-derived cyclooxygenase-2 (COX-2) and its product, prostaglandin E2, exert strong immunoinhibitory effects that block dendritic cell function and CD4+ and CD8+ T-cell proliferation and function. We have shown previously that the addition of an oral COX-2 inhibitor to immunogene therapy using IFN-beta markedly augmented therapeutic efficacy in murine tumor models. In this study, we hypothesized that COX-2 inhibition might also augment an antitumor vaccination strategy. Mice bearing tumors derived from TC1 cells, a tumor line that expresses the human papillomavirus (HPV) E7 protein, were thus vaccinated with an adenoviral vector expressing HPV E7 protein (Ad.E7). This vaccine approach effectively generated E7-specific CD8+ cells and slowed the growth of small tumors but had little effect on large tumors. However, feeding mice with the COX-2 inhibitor, rofecoxib, restored the effectiveness of the vaccine against large tumors and prolonged survival. This effect was accompanied by a larger percentage of E7-specific CD8+ cells in the regional draining lymph nodes and a markedly increased number of tumor-infiltrating E7-specific CD8+ cells (as determined by flow cytometry) and total CD8+ T cells (as determined by immunohistochemical staining). Increased immunocyte trafficking was likely mediated by the generation of a Th1-type tumor microenvironment because COX-2 inhibition increased expression levels of mRNA for IFN-gamma, interleukin-12, IP-10, and MIG while lowering the expression of vascular endothelial growth factor within tumors. This study shows that the effectiveness of a cancer vaccine can be significantly improved by adding COX-2 inhibition.     

Inverse correlation between tumoral indoleamine 2,3-dioxygenase expression and tumor-infiltrating lymphocytes in endometrial cancer: its association with disease progression and survival.

PURPOSE: Tumor escape from host immune systems is a crucial mechanism for disease progression. We recently showed that the immunosuppressive enzyme indoleamine 2,3-dioxygenase (IDO) is a prognostic indicator for endometrial cancer. The purpose of the present study was to investigate the relationship between IDO expression and tumor-infiltrating lymphocytes (TIL) or natural killer (NK) cells and to clarify their prognostic effect in endometrial cancer. EXPERIMENTAL DESIGN: Immunohistochemical staining for IDO expression in endometrial cancer tissues (n = 65) was done. Tumor-infiltrating CD3+ and CD8+ lymphocytes, as well as CD57+ NK cells, were counted in serial tissue sections. RESULTS: High IDO expression in tumor cells was found in 32 of 65 cases and was positively correlated with myometrial invasion, nodal metastasis, and lymph-vascular space involvement. We also found a significant correlation between high IDO expression and reduced numbers of CD3+, CD8+, and CD57+ cells infiltrating into both the tumor epithelium and stroma. Patients with high IDO expression, a low number of stromal CD3 (<60), low intraepithelial CD8 (<25), or low stromal CD8 (<40) had significantly impaired progression-free survival. On multivariate analysis, IDO expression and the number of stromal CD3+ TILs were independent prognostic factors for impaired progression-free survival. CONCLUSIONS: Tumoral IDO expression correlated with a reduced number of TILs and NK cells in endometrial cancer, possibly contributing to disease progression and impaired clinical outcome. These findings suggest that targeting IDO to restore host antitumor immunity may be a therapeutic strategy for endometrial cancer.     

碳离子(^(12)C^(6+))抑制JAK2/STAT3通路促进肺癌中CD8^(+)T细胞浸润的研究北大核心CSCD

目的探索碳离子(^(12)C^(6+))照射后JAK2/STAT3通路的改变及下游蛋白FOXP3调控的肺癌中CD8+T细胞的浸润差异。方法基于C57BL/6小鼠Lewis荷瘤模型的RNA测序分析,筛选出碳离子照射后肺癌中显著改变的JAK2/STAT3通路及相关的差异表达基因及蛋白如FOXP3。利用R软件“GSVA”中ssGSEA免疫浸润算法,探索FOXP3与肺癌免疫微环境中主要免疫细胞浸润的相关性并基于碳离子联合STAT3抑制途径(氯硝柳胺)对肺癌中CD8+T细胞浸润进行分析。结果碳离子照射后,肺癌中JAK2/STAT3通路被抑制,相关基因和蛋白表达下调。基于ssGSEA算法的免疫评分显示,FOXP3表达与肺癌免疫微环境中CD8+T细胞浸润呈显著负相关。通过碳离子照射联合STAT3抑制剂氯硝柳胺,进一步明确了靶向JAK2/STAT3通路对于增加肺癌中CD8^(+)T细胞浸润的协同作用。结论碳离子(^(12)C^(6+))可以通过靶向JAK2/STAT3通路与免疫治疗发挥协同增效的作用。     

碳离子（12C6+）抑制JAK2/STAT3通路促进肺癌中CD8+ T细胞浸润的研究

目的 探索碳离子（¹²C⁶⁺）照射后JAK2/STAT3通路的改变及下游蛋白FOXP3调控的肺癌中CD8⁺T细胞的浸润差异。方法 基于C57BL/6小鼠Lewis荷瘤模型的RNA测序分析,筛选出碳离子照射后肺癌中显著改变的JAK2/STAT3通路及相关的差异表达基因及蛋白如FOXP3。利用R软件“GSVA”中ssGSEA免疫浸润算法,探索FOXP3与肺癌免疫微环境中主要免疫细胞浸润的相关性并基于碳离子联合STAT3抑制途径（氯硝柳胺）对肺癌中CD8⁺T细胞浸润进行分析。结果 碳离子照射后,肺癌中JAK2/STAT3通路被抑制,相关基因和蛋白表达下调。基于ssGSEA算法的免疫评分显示,FOXP3表达与肺癌免疫微环境中CD8⁺T细胞浸润呈显著负相关。通过碳离子照射联合STAT3抑制剂氯硝柳胺,进一步明确了靶向JAK2/STAT3通路对于增加肺癌中CD8⁺T细胞浸润的协同作用。结论 碳离子（¹²C⁶⁺）可以通过靶向JAK2/STAT3通路与免疫治疗发挥协同增效的作用。     

Identification of HLA-A3-restricted CD8+ T cell epitopes derived from mammaglobin-A,a tumor-associated antigen of human breast cancer

Mammaglobin-A is highly overexpressed in breast cancer cell lines and primary breast tumors. This pattern of expression is restricted to mammary epithelium and metastatic breast tumors. Thus, mammaglobin-A-specific T cell immune responses may provide an important approach for the design of breast cancer-specific immunotherapy. The purpose of our study was to define the T cell-mediated immune response to mammaglobin-A. We determined that the frequency of mammaglobin-A-reactive CD8+ and CD4+ T cells in breast cancer patients is significantly higher than that observed in healthy female controls using limiting dilution analyses (p = 0.026 and p = 0.02, respectively). We identified 8 mammaglobin-A-derived 9-mer peptides with the highest binding affinity for the HLA-A3 molecule (Mam-A3.1-8) using a computer-assisted analysis of the mammaglobin-A protein sequence. Subsequently, we determined that CD8+ T cells from breast cancer patients reacted to peptides Mam-A3.1 (23-31, PLLENVISK), Mam-A3.3 (2-10, KLLMVLMLA), Mam-A3.4 (55-63, TTNAIDELK) and Mam-A3.8 (58-66, AIDELKECF) using an IFN-gamma enzyme-linked immunospot assay. A CD8+ T cell line generated in vitro against HLA-A*0301-transfected TAP-deficient T2 cells loaded with these peptides showed significant cytotoxic activity against the Mam-A3.1 peptide. This CD8+ T cell line showed a significant HLA-A3-restricted cytotoxic activity against mammaglobin-A-positive but not mammaglobin-A-negative breast cancer cells. In summary, our study identified four HLA-A3-restricted mammaglobin-A-derived epitopes naturally expressed by breast cancer cells, indicating the immunotherapeutic potential of this novel antigen for the treatment and prevention of breast cancer.     

肝细胞性肝癌组织浸润淋巴细胞表型与分布研究

目的:肝细胞性肝癌组织浸润淋巴细胞与外周血T 细胞表型可能与肿瘤进展及预后相关,本研究检测肝癌患者组织及外周血T 细胞表型与分布,分析淋巴细胞表型变化与预后的关系。方法:分析2007年10月至12月中山医院147 例肝癌及癌旁组织浸润淋巴细胞表型（T 细胞或B 细胞表面标志物:CD3、CD8、CD4、CD20、CD19、Foxp 3）,表型与临床病理特征及预后的关系;检测26例肝癌外周血CD3、CD8、CD4 +T细胞数量并其比例变化。结果:癌巢内肿瘤浸润细胞明显少于癌周组织（P < 0.01）,癌周淋巴细胞主要分布于癌旁正常肝组织、汇管区,其与患者肝炎病史及肝硬化相关,表型以CD3 +T细胞为主,其中又以CD8 + 细胞毒性T 细胞为主;CD4 染色在多数病例为阴性,Foxp 3 仅在个别病例（15/ 109）呈阳性。肿瘤浸润淋巴细胞B 细胞标志CD20、CD19均为阴性。肿瘤组织内CD8 +T细胞浸润数量与预后正相关,而癌周浸润淋巴细胞数目与患者转移及复发无显著关系。结论:肝癌肿瘤浸润细胞在癌巢内明显少于癌周组织,肿瘤及癌周浸润细胞以CD8 + 细胞毒性T 细胞为主。肿瘤组织内CD8 +T细胞浸润数量与预后相关,而癌周浸润淋巴细胞数量与患者转移及复发无显著关系。      

Expression of cyclooxygenase-2 in uterine endometrial cancer and anti-tumor effects of a selective COX-2 inhibitor

A role for cyclooxygenase-2 (COX-2) in the development and progression of various tumors has been identified. Selective COX-2 inhibitors produce anti-proliferative effects in various cancer cell lines that express COX-2. However, the mechanisms underlying anti-tumor effects are unclear. Furthermore, few studies have studied COX-2 expression in gynecological cancers, especially endometrial cancer. The current study had two goals. We investigated the correlation between COX-2 expression and clinicopathological factors of uterine endometrial cancer. We also investigated effects of treatment with etodolac, a selective COX-2 inhibitor, on the uterine endometrial cancer cell line TMG-L, which expresses COX-2. We conclusively confirmed expression of COX-2 mRNA and protein in endometrial cancer that exceeded levels of COX-2 seen in normal endometrium. However, no significant correlations were observed between COX-2 expression in endometrial cancer tumor samples and clinicopathological factors or disease-free survival rate of patients with endometrial cancer. Study of COX-2 inhibition of TMG-L cells showed that etodolac produced dose-dependent inhibition of cell proliferation through G1 phase cell-cycle arrest. Etodolac-induced cell-cycle arrest might be caused by increases in p53 and P21WAF1 protein expression. Production of basic-fibroblast growth factor (bFGF, a pro-angiogenesis factor) was inhibited by etodolac in a dose-dependent manner. Furthermore, telomerase activity was inhibited and expression of hTERT mRNA was significantly inhibited with etodolac, leading to the conclusion that anti-tumor effects of etodolac on TMG-L cells are due to inhibition of both angiogenesis and telomerase activity. These results strongly suggest that COX-2 inhibitors have potential as therapeutic (and possibly, chemopreventive) agents for endometrial cancers that overexpress COX-2.     

PD-L1与PD-1在宫颈鳞癌与宫颈良性病变组织及外周血中的表达及其预后价值

目的:通过分别对宫颈鳞癌、宫颈良性疾病组织和配对血液标本检测,了解外周血程序性死亡受体1(PD-1)与肿瘤组织程序性死亡受体-配体1(PD-L1)表达的一致性,初步评价其与宫颈鳞癌预后的关系。方法:收集宫颈鳞癌及宫颈良性病变组织标本各70例,并收集配对的外周血标本鳞癌24例,良性病变30例。免疫组化法检测组织PD-L1,流式细胞术检测外周血T细胞表面PD-1+亚群的比例,并结合生存资料做统计分析。结果:宫颈鳞癌患者外周血PD-1与组织PD-L1表达呈正相关（r2=0.734,P=0.000;r2=0.66,P=0.000）,宫颈良性病变者外周与组织的表达不相关（r2=0.138,P=0.043;r2=0.174,P=0.022）。宫颈癌患者血CD8+PD-1+细胞百分比、组织PD-L1+的细胞数大于宫颈良性病变组（P=0.000,P=0.002）。宫颈癌患者随肿瘤直径增大、分期升高,肿瘤组织PD-L1表达量也相应升高,外周T细胞表面PD-1表达也有类似趋势。但是外周T细胞表面PD-1表达程度与肿瘤大小、分化程度、深肌层侵犯均无关。单因素分析提示外周CD4+T细胞低表达PD-1的、组织低表达PD-L1的患者生存期较长。多因素分析结果显示,PD-L1表达对患者生存有影响（B=1.844,P=0.019）。结论:宫颈鳞癌患者外周血PD-1、组织PD-L1表达与肿瘤的分级、预后有关,组织PD-L1表达是患者的独立预后因素。