Expression of Thymidine Phosphorylase in Human Gastric Carcinoma

The activity of thymidine phosphorylase (dThdPase) has heen reported to increase in several types of malignant tumors. Experimental evidence has shown that dThdPase is identical to platelet-derived endothelial cell growth factor, and that dThdPase has angiogenic activity. We examined the expression of dThdPase to investigate whether the expression of dThdPase correlates with angiogenesis, clinicopathologic features and the prognosis of patients with human gastric carcinomas. Microvessels were assessed by immnnostaining endothelial cells for factor VIII. We counted microvessels in the tumors of 158 patients whose tumors were completely removed surgically. Microvessels were counted in a × 400 field in the most active areas of neovascularization. We purified a monoclonal antibody (TMA-1) against dThdPase and studied the expression of dThdPase using TMA-1 in the same serial sections as those used for the detection of factor VIII. The correlation between angiogenesis and dThdPase, and the clinicopathological significance of dThdPase, in patients with gastric carcinoma were examined. The positive expression of dThdPase was more frequent (P<0.001) in gastric carcinomas (67/158, 43.4%) than that in normal tissues (12/158, 7.6%). The average microvessel count in dThdPase-positive gastric carcinomas was higher (P<0.001) than that in dThdPase-negative carcinomas. The percentage of gastric carcinoma cells expressing dThdPase was significantly correlated with the microvessel count (P<0.001). Further, the average size of dThdPase-positive carcinomas was significantly larger (P<0.001) than that of negative carcinomas and the mean microvessel count in dThdPase-positive gastric carcinomas was also significantly higher (P<0.001) than that in dThdPase-negative carcinomas. There was a significant correlation between the positive expression of dThdPase and microvessel count (P<0.001) or lymph node metastasis (P=0.013) by multivariate logistic analysis. Further, patients with dThdPase-positive carcinoma showed a significantly worse prognosis than those with dThdPase-negative carcinoma overall and in stage III. These findings indicate that the expression of dThdPase in gastric carcinomas is related to progression and metastasis, and this enzyme affects the prognosis of some patients with the disease.     

子宫内膜癌组织中胸苷磷酸化酶的表达及临床意义

目的：探讨子宫内膜癌病人胸苷磷酸化酶（TP／PD－ECGF）表达与内膜癌血管生成、临床病理特征及预后的关系。方法：随机选取子宫内膜腺癌石蜡标本48例，其中I期36例、Ⅱ期8例、Ⅲ期4例（FIGO分期），绝经者28例。不典型增生石蜡标本22例，正常内膜石蜡标本15例。用TP／PD－ECGF单抗免疫组化染色确定组织内的TP表达，用CD34单抗免疫组化染色确定组织内微血管，然后分析内膜癌TP表达与微血管计数（MVD）以及TP表达和MVD分别与临床病理特征（绝经状态、组织学分级、临床分期、肌层浸润、宫颈浸润、淋巴结转移）、与无病生存时间的关系。结果：正常内膜组80％病例TP在腺体细胞表达，而不典型增生组及内膜腺癌组TP以基质及基质和腺体同时表达为主（分别为91％、78．2％），正常组与不典型增生组及内膜腺癌组比较，差异有显著性（P＜0．005）。不典型增生组与内膜癌组比较，TP表达无显著差异（P＞0．1）。不典型增生组MVD（平均为32．7），较正常的内膜组MVD（平均为27．1）高（P＜0．01），但明显较内膜腺癌组MVD（平均为78．9）低（P＜0．001），子宫内膜癌TP基质＋癌染色组MVD（平均值79．1）低于基质染色组MVD（平均值86．2，P＜0．05），但明显高于癌细胞染色组MVD（平均值60．5P＝0．005）。子宫内膜腺癌TP表达及MVD均与临床病理特征无关，基质TP染色及高MVD的病人无病生存期短。结论：子宫内膜癌TP表达与临床病理特征无关，而与MVD及预后有关。TP在内膜癌血管生成中起重要作用，TP和MVD一样能预测内膜癌病人的预后。血管生成与子宫内膜癌发生、发展密切相关，TP可能与内膜癌的发生有密切的关系。     

Heme Oxygenase-1 Inhibits Tumor Metastasis Mediated by Notch1 Pathway in Murine Mammary Carcinoma

Heme oxygenase-1 (HO-1) plays an important role in the progression of several malignancies including breast cancer. However, its role in breast cancer metastasis is still ambiguous. In this study, we observed the effect of HO-1 on mouse mammary carcinoma metastasis using the in vivo tumor metastasis model. Our results revealed that overexpression of HO-1 strongly inhibits the lung metastasis of 4T1 cells. In in vitro analysis, associated indices for epithelial–mesenchymal transition (EMT), migration, and proliferation of 4T1 cells were evaluated. The results show that HO-1 inhibits EMT, migration, and proliferation of 4T1 cells. In addition, the Notch1/ Slug pathway is found to mediate an antimetastasis role of HO-1 in mouse mammary carcinoma. In conclusion, since HO-1/Notch1/Slug axis plays an important role in breast cancer metastasis, induction of HO-1 could be used as a potential therapeutic strategy for breast cancer treatment.     

Significance of Thymidine Phosphorylase/Platelet-derived Endothelial Cell Growth Factor in Carcinoma of the Papilla of Vater

The expression of thymidine phosphorylase (TP) in carcinoma of the papilla of Vater was studied to clarify its significance in tumor progression and in determining prognosis. Fifty-nine cases of surgically resected carcinoma of the papilla of Vater were studied. Immunohistochemical staining was performed to evaluate the expression of TP, microvessel count and p53 overexpression. TP expression was demonstrated in tumor cells in 62.7% (37/59) of the cases. A higher frequency of regional lymph node metastasis was found in TP-positive tumors than in TP-negative tumors ( P =0.006). TP-positive tumors were more advanced than TP-negative tumors with regard to clinical stage ( P =0.035). TP-positive tumors had significantly higher microvessel density (27.6±10.1) than TP-negative tumors (20.4±10.0, P =0.01). Moreover, TP expression was significantly correlated with a poor prognosis ( P =0.02). These suggest that in carcinoma of the papilla of Vater, TP production by tumor cells is correlated with tumor progression through its regulatory effect on neovascularization.     

Suppression of Tumor Growth and Downregulation of Platelet-derived Endothelial Cell Growth Factor/Thymidine Phosphorylase in Tumor Cells by Angiogenesis Inhibitor TNP-470

We investigated the effects of the angiogenesis inhibitor TNP-470 on human lung squamous cell carcinoma cell lines H226B and H226Br both in vivo and in vitro . H226B was established from human lung squamous cell carcinoma and H226Br was established from a brain metastatic lesion of H226B in nude mice. Nude mice inoculated with these cells were treated with 30 mg/kg of TNP-470 subcutaneously every other day. At this dose, TNP-470 only significantly suppressed the growth of H226Br tumor, but not H226B tumor. Attempts to use a high dose of TNP-470 (100 mg/kg) resulted in a severe loss of body weight. Immunohistochemical studies showed marked tumor vascularization in H226Br tumor, but the formation of new blood vessels was suppressed by 30 mg/kg of TNP-470. Investigation of the mechanism of anti-angiogenic effects of TNP-470 in vivo showed that the expression and the activity of platelet-derived endothelial cell growth factor/thymidine phosphorylase (PD-ECGF/dThdPase) in H226Br tumor was significantly suppressed by 30 mg/kg of TNP-470. Furthermore, TNP-470 inhibited cell growth of cultured H226Br dosedependently at concentrations of ≥1 μg/ml. Immunoblot analysis revealed H226Br cells gave a stronger PD-ECGF signal than H226B cells, and the expression of PD-ECGF/dThdPase in H226Br was also suppressed by treatment with TNP-470 at ≥0.1 μg/ml. No change in basic fibroblast growth factor (bFGF) or vascular endothelial growth factor (VEGF) was noted in these cell lines. Our results suggested that TNP-470 acts, at least in part, by downregulation of PD-ECGF/ dThdPase in this cell line.     

血红素加氧酶-1与肿瘤的研究进展

血红素加氧酶-1(HO-1)是一种重要的抗炎、抗凋亡、抗氧化及耐药相关基因。在生理状态下,人体正常组织中的HO-1蛋白呈低表达。在遭受酒精、辛辣及热烫等持续刺激时,其表达升高。HO-1的表达异常与肿瘤的生物学行为密切相关。近年来研究发现,其基因多态性及分解血红素产生的代谢产物(胆红素、Fe²⁺、CO)参与的多条信号通路的激活,并影响多种肿瘤疾病的发生、发展。另外,在许多肿瘤中HO-1表达水平上调,不仅影响肿瘤微环境中氧化应激水平对肿瘤细胞生长及增殖的调节,而且还能诱导肿瘤细胞产生耐药性。因此,HO-1表达异常可能参与了肿瘤的发生发展。     

PD-1/PD-L1抑制剂在恶性黑色素瘤（新）辅助治疗中的研究进展

恶性黑色素瘤是一种具有较高复发率及死亡率的实体肿瘤。目前针对Ⅱ～Ⅲ期黑色素瘤的治疗仍是以外科手术为主,由于缺乏有效的(新)辅助治疗手段,高危黑色素瘤患者的5年生存率仅有30%～70%。程序性死亡因子1(PD-1)和程序性死亡因子配体1(PD-L1)是重要的免疫检验点共抑制分子,通过抑制T细胞的激活和增殖通路参与了肿瘤的免疫逃逸。近些年来包括Nivolumab和Pembrolizumab在内的多种PD-1/PD-L1抑制剂研发获批上市,并在晚期黑色素瘤的治疗中表现出显著的疗效。因此多项针对PD-1/PD-L1抑制剂在高危黑色素瘤患者(新)辅助治疗的临床试验正积极开展。本文主要对近年来相关研究进行了回顾与总结,探讨PD-1/PD-L1抑制剂在黑色素瘤(新)辅助治疗应用的前景和可能发展的方向。     

RETRACTED ARTICLE: Poly(Adenosine Diphosphate-Ribose) Polymerase-1 Expression in Cutaneous Malignant Melanomas as a New Molecular Marker of Aggressive Tumor

Poly(adenosine diphosphate-ribose) polymerases (PARPs) are a family of enzymes, which catalyses poly (ADP-ribosyl)ation of DNA-binding proteins and directly involved in genomic stability, DNA repair, and apoptosis. In this study, we evaluated the immunomorphology of PARP-1 in melanoma and its prognostic importance. We studied PARP-1 expression by immunohistochemistry in a selected series of 54 primary cutaneous malignant melanoma (CMM). The findings of the present study suggest that the neoplastic progression toward the invasive (both horizontal and vertical) growth phase of CMM cells is characterized by the loss of cleavage of PARP-1, probably signaling an imbalance of the apoptotic process in these cells and leading to further gain to aggression. Over-expression of full-length PARP-1 was correlated with recurrence and/or progression of the disease and so act as a promising new biological marker of CMM. Our study represents the evidence of a direct correlation between the PARP-1-mediated apoptotic process and the biologic behavior of CMM.     

水通道蛋白1及其mRNA在胸膜恶性肿瘤进展中的作用

目的 探讨胸膜恶性肿瘤的进展对水通道蛋白1（aquaporin-1,AQP1）及其mRNA的影响。方法 通过胸膜腔注射法建立胸膜恶性肿瘤的动物模型,按肿瘤种植时间分组（T6,T8,T10）,记录各组胸水量,采用免疫组织化学法、Real-Time PCR的方法测定AQP1及AQP1-mRNA水平。结果 肿瘤小鼠胸水量随时间而增长;壁层胸膜AQP1及其mRNA水平随肿瘤进展而增高,且与胸水量呈正相关。结论 在胸膜恶性肿瘤的进展中,胸膜壁层AQP1及其mRNA水平升高,胸水量与AQP1、AQP1-mRNA水平呈正相关,进而提示AQP1参与了胸膜恶性肿瘤的构建和进展。     

人端粒酶催化亚单位及其相关蛋白在骨肿瘤中的表达

目的：研究骨肿瘤中端粒酶相关基因中,人端粒酶催化亚单位（human elomerase catalytic subunit,hTERT)和人端粒酶相关蛋白（human telomerase associated protein,TP1)两个重要成分的表达及对评估肿瘤良、恶性的意义。方法：提取50例手术切除骨肿瘤组织、2种人成骨内瘤细胞系Saos－2、OS732和原代培养人成骨细胞的总RNA。用一步RT－PCR方法检测hTERT和TP1mRNA的表达。结果：所有骨肿瘤标本及细胞系均检测到TP1的表达。hTERT在17例良性骨肿瘤、33例骨肉瘤、2种人成骨肉瘤细胞系和人成骨细胞的表达率分别为64．7％、57．6％、0％和0％。良、恶性肌肿瘤间hTERT表达的差异不具有显著性（P＞0．05）,不同分化程度骨肉瘤间hTERT表达的差异也不具有显著性（P＞0．05）。结论：一步RT－PCR是检测端粒酶亚单位的有效方法。hTERT和TP1表达水平的上调在骨肿瘤恶性进展中可能不起主要作用,在骨肿瘤发展和维持中可能有替代性（ALT）机制导致细胞永生化。     

TRP-1反义核酸抑制恶性黑素瘤细胞增殖的体外及体内研究

目的:观察TRP-1反义核酸在体内、外对恶性黑素瘤细胞增殖的抑制作用,探讨黑素瘤治疗的新途径.方法:构建TRP-1反义核酸真核表达载体,将其转染恶性黑素瘤细胞,以测定的MTT结果绘制细胞生长曲线;进行黑素瘤细胞体外克隆形成实验观察,计算各组细胞的克隆形成率;以TRP-1反义核酸转染的黑素瘤细胞及对照组细胞注射裸鼠,观察肿瘤大小及增长速度.结果:从生长曲线可以看出,TRP-1反义核酸转染的黑素瘤细胞增殖速度明显低于对照细胞;体外克隆形成实验结果显示:TRP-1反义核酸转染的黑素瘤细胞克隆形成率为52%,而对照组为68%(P＜0.01);裸鼠成瘤试验表明,转染TRP-1反义核酸的细胞成瘤性显著低于未转染细胞及转染载体对照细胞(P＜0.01).结论:TRP-1反义核酸在体内、外均能明显抑制恶性黑素细胞的增殖,在恶性黑素瘤的治疗中有一定的应用前景.     

桑黄灵芝UE-1对肿瘤生长及血管新生的抑制作用

目的观察桑黄灵芝UE-1对Lewis肺癌生长及血管新生的抑制作用。方法建立Lewis肺癌实体瘤模型,观察其抗肿瘤作用;通过免疫组织化学染色,检测肿瘤微血管密度;采用新生血管计数实验检测UE-1对肿瘤组织血管生成的影响;通过鸡胚尿囊膜血管新生实验观察UE-1多糖的抗血管新生作用。结果实体瘤模型中,UE-1组移植瘤重量和体积明显低于对照组（P<0.05）,且微血管密度值也低于对照组（P<0.05）;肿瘤诱导新生血管中UE-1组肿瘤周围的血管数明显少于对照组（P<0.05）;鸡胚尿囊膜血管新生实验显示UE-1多糖具有抑制血管新生作用。结论桑黄灵芝UE-1可抑制Lewis肺癌的生长,可能是通过抑制肿瘤血管新生来发挥其抑瘤作用。     

低氧对恶性黑素瘤A375细胞中Nodal及细胞迁移相关蛋白表达的影响

目的 探讨低氧对恶性黑素瘤A375细胞中Nodal及细胞迁移相关蛋白表达的影响.方法 对恶性黑色素瘤A375细胞进行体外培养,细胞分为空白对照组、CoCl2组、SB-431542阻断组和CoCl2+SB-431542组,以氯化钴(CoCl2)诱导构建A375细胞内的低氧环境,SB-431542阻断Nodal信号通路,应...     

Radixin Knockdown by RNA Interference Suppresses Human Glioblastoma Cell Growth in Vitro and in Vivo

Radixin, a member of the ERM (ezrin–radixin–moesin) family, plays important roles in cell motility, invasionand tumor progression. It is expressed in a variety of normal and neoplastic cells, including many types ofepithelial and lymphoid examples. However, its function in glioblastomas remains elusive. Thus, in this study,radixin gene expression was first examined in the glioblastoma cells, then suppressed with a lentivirus-mediatedshort-hairpin RNA (shRNA) method.We found that there were high levels of radixin expression in glioblastomaU251cells. Radixin shRNA caused down-regulation of radixin gene expression and when radixin-silenced cellswere implanted into nude mice, tumor growth was significantly inhibited as compared to blank control cells or nonsenseshRNA cells. In addition, microvessel density in the tumors was significantly reduced. Thrombospondin-1(TSP-1) and E-cadherin were up-regulated in radixin- suppressed glioblastoma U251 cells. In contrast, MMP9was down-regulated. Taken together, our findings suggest that radixin is involved in GBM cell migration andinvasion, and implicate TSP-1, E-cadherin and MMP9 as metastasis-inducing factors.     

Immunoscintigraphy and Pharmacokinetics of Indium-111-labeled ZME-018 Monoclonal Antibody in Patients with Malignant Melanoma

Immunoscintigraphic and pharmacokinetic characteristics of ¹¹¹In-labeled ZME-018 monoclonal antibody were examined in 8 patients with malignant melanoma. Each patient received a single intravenous infusion of 20 mg of ZME-018, coupled to 3 mCi of ¹¹¹In without any acute toxicity. Scintigrams were taken 1, 3, and 6 days after the administration, and blood and urine samples were also taken frequently. Rapid clearance of some radioactivity was seen in early urine samples in the form of ¹¹¹In DTPA, but after 1 day, urinary excretion of radioactivity was slow and steady, with an average of 2.5% of the injected dose excreted per day. The scans demonstrated that there was blood retention of radioactivity in the heart and great vessels 1 day after infusion and considerable clearance from the blood pool occurred by 3 days. However, ¹¹¹In was deposited in the liver, spleen and bone for up to 6 days. The optimal time for imaging appeared to be at 3 days. Nineteen out of 26 known lesions or 6 out of 8 patients were positive. There were 21 lesions detected that were not suspected during the work-up the patient. Five patients developed human anti-mouse antibody in the serum by 3 weeks. These results suggest that immunoscintigraphy with ¹¹¹In-labeled ZME-018 antibody is safe and useful for the detection of metastatic lesions in a selected group of patients with malignant melanoma.