肿瘤坏死因子-α在卵巢肿瘤患者腹水中的表达及意义

目的 探讨卵巢肿瘤患者腹水中肿瘤坏死因子-α(TNF-α)与卵巢癌之间的关系,探讨其对腹水性质鉴别的价值.方法 采用酶联免疫吸附法(ELISA)检测59例卵巢上皮性肿瘤、9例原发性腹膜癌、6例卵巢子宫内膜异位囊肿患者腹水TNF-α水平.结果 腹水TNF-α水平在卵巢恶性上皮性肿瘤、卵巢交界性肿瘤、原发性腹膜癌中的水平明显高于卵巢良性上皮性肿瘤、卵巢子宫内膜异位囊肿,且与肿瘤的恶性行为包括临床分期、组织分化程度、淋巴转移及腹水量等有关.结论 腹水TNF-α对诊断卵巢癌有较高的敏感性和特异性,可能成为卵巢恶性肿瘤的标志物.     

卵巢癌组织中血管生成拟态观测及其与临床病理特征和预后的关系

目的 探讨卵巢癌血管生成拟态这一独特的肿瘤营养供应模式及其与卵巢癌临床病理特征和预后的关系.方法 收集临床和随访资料完整的卵巢癌患者经手术切除的石蜡标本共84例,分析卵巢癌的临床病理特征;采用CD31/PAS双重染色法将84例卵巢癌组织分为有血管生成拟态组和无血管生成拟态组,分析血管生成拟态与患者临床病理特征及肿瘤转移和预后之间的关系;并采用免疫组织化学(SP法)检测两组卵巢癌组织中血管内皮生长因子(VEGF)、基质金属蛋白酶(MMP)-2、MMP-9、E-cadherin、β-catenin和波形蛋白的表达情况.结果 84例卵巢癌患者中有36例存在血管生成拟态,84例卵巢癌的FIGO分期、组织学类型、病理学分级和转移情况与血管生成拟态的形成密切相关(X2值分别为10.30、20.01、8.16、9.75,P值均小于0.05).VEGF、MMP-2、MMP-9、E-cadherin和β-catenin在血管生成拟态组卵巢癌组织中呈高表达状态,在无血管生成拟态组组织中呈低表达状态.生存分析发现有血管生成拟态的患者生存时间比无血管生成拟态患者短(P=0.04).结论 卵巢癌细胞形成血管生成拟态的能力与其恶性度有关,恶性度越高形成血管生成拟态的能力越强,血管生成拟态是卵巢癌患者预后的重要指标,VEGF、MMP-2、MMP-9与卵巢癌血管生成拟态的形成有着密切的关系.     

吲哚胺2,3双加氧酶在上皮性卵巢癌组织中的表达及其与预后相关性的研究

目的:研究吲哚胺2,3双加氧酶(IDO)在上皮性卵巢癌组织中的表达及其与预后的相关性。方法:通过免疫组化的方法测定IDO在卵巢良性上皮性肿瘤、交界性上皮性肿瘤和恶性上皮性肿瘤组织中的表达。结果:IDO在卵巢恶性肿瘤中表达显著高于卵巢良性肿瘤(P<0.05)。IDO的表达与组织学分级显著相关(P<0.05)。IDO的阳性表达在复发和死亡组中明显高于生存组(P<0.05)。IDO(+)者的生存率曲线、平均生存时间明显低于IDO(-)者(P<0.05)。结论:低分化、高度恶性的卵巢癌细胞更可能表达IDO,IDO阳性表达与卵巢癌不良预后显著相关。     

卵巢上皮肿瘤组织Fas、Angs、E-cad的表达水平及意义分析

目的检测Fas、促血管生成素-1(Ang-1)、促血管生成素-2(Ang-2)、E-钙粘蛋白(E-cad)在卵巢上皮肿瘤组织中的表达水平,藉此评价其与肿瘤发生发展及预后的关系。方法应用免疫组织化学S-P法和原位杂交技术,检查了21例卵巢上皮恶性肿瘤,18例良性肿瘤,8例正常卵巢组织中Fas,Ang-1,Ang-2及E-cad的表达。结果 Fas的表达随肿瘤病变程度的增高而逐步减弱,恶性肿瘤组织与正常卵巢及良性肿瘤组织间有显著差异(P<0．05);E-cad免疫反应阳性强度在卵巢上皮癌组织中明显降低,与正常卵巢及良性肿瘤组织间有显著性差异(P<0．05)。原位杂交显示,Ang-1 mRNA在正常卵巢及肿瘤组织中都有表达,其表达水平没有显著差异(P>0．05);Ang-2 mRNA在恶性肿瘤组织中表达明显上调(P<0．01);E-cad mRNA在恶性肿瘤组织中表达明显下调(P<0.01)。结论 Fas的表达水平与卵巢上皮肿瘤的内在发展相关;由E-cad介导的细胞间黏附作用的减弱是卵巢癌发生、发展及转移的一个重要因素;Ang-2对于促进卵巢癌组织血管新生和形成可能具有重要促进作用。     

卵巢癌干细胞在腹腔温热化疗治疗卵巢癌腹膜转移及并发恶性腹水中的作用

卵巢癌是女性最常见的恶性肿瘤之一,腹膜转移及并发的恶性腹水是影响卵巢癌预后的主要因素.研究发现,癌症的复发或转移由癌症干细胞(cancer stem cells,CSC)引起,针对CSC而不是成熟的癌细胞的治疗策略在癌症腹膜转移及恶性腹水的治疗中起至关重要的作用[1].本文仅对卵巢癌CSC在腹腔温热化疗(intraperitoneal hyperthemic chemotherapy,IHPC)治疗卵巢癌腹膜转移及并发恶性腹水中的作用作一简要综述.     

垂体肿瘤转化基因、整合素连接激酶和迁移诱导蛋白7在卵巢上皮性肿瘤中的表达及意义

目的:探讨垂体肿瘤转化基因(PTTG)、整合素连接激酶(ILK)和迁移诱导蛋白7(Mig-7)在卵巢上皮性肿瘤发生发展过程中的表达及意义.方法:收集天津市肿瘤医院临床、病理和预后资料完整的恶性卵巢肿瘤组织标本76例,交界性卵巢肿瘤20例,良性卵巢肿瘤组织和正常卵巢组织各10例,进行PTTG、ILK和Mig-7免疫组织化学显色,参考Fromowitz 方法判定统计免疫组织化学显色结果.结果:恶性卵巢肿瘤中PTTG、ILK和Mig-7的表达与交界性、良性卵巢肿瘤及正常卵巢组织中表达的差异有统计学意义；PTTG、ILK和Mig-7的阳性表达在不同组织学分级(I～Ⅱ级,Ⅲ～Ⅳ级)、有或无腹水、有或无远处转移和有或无淋巴结转移组别病例巾表达的差异有统计学意义.结论:PTTG、ILK和Mig-7在卵巢上皮性恶性肿瘤的发生发展中起重要作用,可以作为临床判断卵巢癌生物学行为的分子生物学指标.     

CK7和CA125在人卵巢癌的表达及意义

目的研究细胞角蛋白7(CK7)和CA125联合在卵巢恶性肿瘤中的诊断意义及价值。方法采用免疫组化方法,分别检测100例卵巢肿瘤中CK7和CA125的表达,统计学方法比较两种指标在良性、恶性及转移性肿瘤中的阳性表达率。结果 CK7和CA125在卵巢恶性肿瘤中的表达水平均高于转移源性的卵巢癌和卵巢良性瘤,差异均有统计学意义(0.05)。且在晚期癌中的表达高于早期,差异均有意义(0.05)。结论CK7和CA125过表达提示二者可能参与卵巢癌的发病机制。     

血管内皮细胞生长因子与肿瘤的研究现状

恶性肿瘤的无限制侵袭性生长及其转移依赖于血管生成.因此,目前阻断血管形成探索抑制肿瘤的新途径之一.而血管内皮细胞生长因子(VEGF)是高度特异性的血管内皮细胞促分裂素,是重要的血管形成因子,从而支持肿瘤的生长.研究血管生成对了解恶性肿瘤发生、发展、侵袭、转移的生物学特性及针对肿瘤血管生成的基因治疗有重要的意义.     

卵巢上皮-间质肿瘤中B7-H3、B7-H4的表达及临床病理意义

目的探讨B7-H3、B7-H4在卵巢上皮-间质肿瘤中的表达及其临床病理意义。方法用RT-PCR技术及免疫组化PV9000两步法研究86例卵巢恶性上皮-间质肿瘤(恶性组),40例卵巢交界性上皮-间质肿瘤(交界组)和40例卵巢良性上皮-间质肿瘤(良性组)B7-H3与B7-H4mRNA及蛋白表达情况,并结合临床病理参数进行分析。结果 B7-H3、B7-H4mRNA在3组卵巢肿瘤组织中均有表达,且各自的3组卵巢肿瘤组织的mRNA光密度值差异均有统计学意义(P<0.05);B7-H3与B7-H4蛋白在恶性组中均呈细胞质和(或)细胞膜表达,其中B7-H3阳性率为81.4%(70/86),明显高于交界组22.5%(9/40)和良性组5.0%(2/40)的表达,差异有统计学意义(P<0.05);B7-H4阳性率为77.9%(67/86),明显高于交界组35.0%(14/40)和良性组5.0%(2/40)的表达,差异有统计学意义(P<0.05)。恶性组的卵巢癌组织中B7-H3蛋白在非黏液性卵巢癌中较黏液性卵巢癌高表达,差异有显著性(P<0.01),与临床分期、病理分级、患者年龄、腹水细胞学和淋巴结转移差异均无统计学意义(P>0.05)。B7-H4蛋白表达与组织病理学类型、临床分期、病理分级、患者年龄、腹水细胞学和淋巴结转移差异均无统计学意义(P>0.05)。结论 B7-H3与B7-H4在卵巢恶性上皮-间质肿瘤表达提示其可能与肿瘤的发生、发展有关,可为卵巢恶性肿瘤的诊断及治疗提供新的依据。     

EphA2和E-钙粘素在卵巢上皮性肿瘤组织中表达及意义

目的 探讨卵巢上皮性肿瘤组织中EphA2和E-钙粘素的表达及其与临床病理特性的关系.方法 采用免疫组织化学SP法检测40例卵巢恶性上皮性肿瘤、20例卵巢良性上皮性肿瘤及10例正常卵巢组织中EphA2及E一钙粘素蛋白的表达情况.结果 在正常卵巢、卵巢上皮性良性肿瘤和卵巢恶性上皮性肿瘤组织中,EphA2蛋白的阳性表达率分别为10.0%、30.0%、65.0%,差异有统计学意义(P<0.01);E-钙粘素蛋白的阳性表达率分别为90.0%、65.0%、45.0%,差异有统计学意义(P<0.05).卵巢恶性肿瘤中EphA2、E-钙粘素蛋白呈负相关(r=-0.495).EphA2和E-钙粘素的蛋白过表达均与卵巢恶性上皮性肿瘤FIGO分期、组织学分级、淋巴结转移有关.结论 EphA2参与卵巢恶性肿瘤的发生发展过程,其表达与卵巢恶性肿瘤的浸润和转移有关.E-钙粘素与EphA2可能存在相互作用.     

Deciphering the signaling events that promote melanoma tumor cell vasculogenic mimicry and their link to embryonic vasculogenesis: Role of the Eph receptors

During embryogenesis, the primordial microcirculation is formed through a process known as vasculogenesis. The term “vasculogenic mimicry” has been used to describe the manner in which highly aggressive, but not poorly aggressive melanoma tumor cells express endothelial and epithelial markers and form vasculogenic‐like networks similar to embryonic vasculogenesis. Vasculogenic mimicry is one example of the remarkable plasticity demonstrated by aggressive melanoma cells and suggests that these cells have acquired an embryonic‐like phenotype. Since the initial discovery of tumor cell vasculogenic mimicry by our laboratory, we have been focusing on understanding the molecular mechanisms that regulate this process. This review will highlight recent findings identifying key signal transduction events that regulate melanoma vasculogenic mimicry and their similarity to the signal transduction events responsible for promoting embryonic vasculogenesis and angiogenesis. Specifically, this review will focus on the role of the Eph receptors and ligands in embryonic vasculogenesis, angiogenesis, and vasculogenic mimicry. Developmental Dynamics 236:3283–3296, 2007. © 2007 Wiley‐Liss, Inc.     

血管内皮生长因子、Eph受体酪氨酸激酶A2、基质金属蛋白酶-2和-9在卵巢癌血管生成拟态中的作用

目的 探讨血管内皮生长因子(VEGF)、Eph受体酪氨酸激酶A2 (EphA2)、基质金属蛋白酶(MMP)-2和MMP-9在卵巢癌血管生成拟态中的作用.方法 收集临床和预后资料完整的卵巢癌组织标本84例,切片经明确诊断后进行CD31和过碘酸-雪夫 (PAS) 双重染色,证实肿瘤组织中存在血管生成拟态,行VEGF、EphA2、MMP-2和MMP-9 免疫组织化学染色,根据染色指数统计免疫组织化学染色结果.结果 有血管生成拟态组(36/84)和无血管生成拟态组卵巢癌组织(48/84)的VEGF、EphA2、MMP-9表达差异有统计学意义,有血管生成拟态组的卵巢癌细胞VEGF、EphA2、MMP-9表达明显高于无血管生成拟态组.但有血管生成拟态组和无血管生成拟态组患者的MMP-2表达差异无统计学意义.结论 在卵巢癌中血管生成拟态和内皮依赖性血管并存,VEGF、EphA2和MMP-9等参与了卵巢癌血管生成拟态的形成.检测VEGF、EphA2和MMP-9可作为预测卵巢癌预后的间接指标.     

Vasculogenic mimicry

The term vasculogenic mimicry describes the formation of fluid-conducting channels by highly invasive and genetically dysregulated tumor cells. Two distinctive types of vasculogenic mimicry have been described. Vasculogenic mimicry of the tubular type may be confused morphologically with endothelial cell-lined blood vessels. Vasculogenic mimicry of the patterned matrix type in no way resembles blood vessels morphologically or topologically. Matrix proteins such as laminin, heparan sulfate proteoglycan, and collagens IV and VI have been identified in these patterns. The patterned matrix anastomoses with blood vessels, and systemically injected tracers co-localize to these patterns. Vasculogenic mimicry of the patterned matrix type has been identified in uveal, cutaneous and mucous membrane melanomas, inflammatory and ductal breast carcinoma, ovarian and prostatic carcinoma, and soft tissue sarcomas, including synovial sarcoma rhabdomyosarcoma, osteosarcoma, and pheochromocytoma. Because the microcirculation of many tumors may be heterogeneous -- including incorporated or co-opted vessels, angiogenic vessels, mosaic vessels, and vasculogenic mimicry of the tubular and patterned matrix types -- therapeutic regimens that target angiogenesis alone may be ineffective against highly invasive tumors that contain patterned matrices. Vasculogenic mimicry provides an opportunity to investigate the interrelationships between the genetically dysregulated invasive tumor cell, the microenvironment, and the malignant switch.     

The role of Twist1 in hepatocellular carcinoma angiogenesis: a clinical study

The epithelial-mesenchymal transition regulator Twist1 has been implicated in tumor invasion, metastasis, and vasculogenic mimicry formation of human hepatocellular carcinoma. However, the relationship between Twist1 expression and endothelium-dependent angiogenesis is not clear. In this study, to investigate the role of Twist1 in hepatocellular carcinoma angiogenesis, we measured the microvessel density by CD31 immunohistochemistry stain and explored the microvessel density as an angiogenesis indicator. The microvessel density in paraffin sections from 97 patients was correlated with Twist1 expression up-regulation. Nuclear relocation was also identified based on immunohistochemistry stain, presenting a significant clinical pattern in hepatocellular carcinoma metastasis and prognosis. Twist1 expression, which is both located in the cytoplasm and relocated into the nucleus, was associated with matrix metalloproteinase 9 up-regulation; matrix metalloproteinase 2 did not appear to present these effects in hepatocellular carcinoma. An assessment of microvessel density could provide an estimate of the degree of angiogenic activity in tissues, as well as its association with Twist1 up-regulated expression. To the best of our knowledge, not only is Twist1 related to metastasis by tumor cells, but vasculogenic mimicry is also significantly related to microvessel density; this process is also associated with matrix metalloproteinase 9 up-regulated expression. This work provides a better understanding of the role of Twist1 in hepatocellular carcinoma angiogenesis and metastasis, suggesting that our findings could represent tumor cell epithelial-mesenchymal transition and endothelium-dependent angiogenesis, as can be seen in hepatocellular carcinoma.     

Molecular determinants of ovarian cancer plasticity

During development, the formation and remodeling of primary vascular networks occurs by vasculogenesis and angiogenesis. Recently, the term "vasculogenic mimicry" has been used by our laboratory and collaborators to reflect the embryonic-like ability of aggressive, but not nonaggressive, melanoma tumor cells to form a pattern of matrix-rich networks (containing channels) surrounding spheroids of tumor cells in three-dimensional culture, concomitant with their expression of vascular cell markers. Ovarian cancer is usually diagnosed as advanced stage disease in most patients when widespread metastases have already been established within the peritoneal cavity. In this study, we explored whether invasive ovarian carcinoma cells could engage in molecular vasculogenic mimicry reflected by their plasticity, compared with their normal cell counterparts. The data revealed that the invasive ovarian cancer cells, but not normal ovarian surface epithelial cells, formed patterned networks containing solid and hollow matrix channels when grown in three-dimensional cultures containing Matrigel or type I collagen, in the absence of endothelial cells or fibroblasts. Immunohistochemical analysis showed that matrix metalloproteinases (MMP)-1, -2, and -9, and MT1-MMP were discretely localized to these networks, and the formation of the networks was inhibited by treatment with MMP inhibitors. Furthermore, the RNase protection assay revealed the expression of multiple vascular cell-associated markers by the invasive ovarian cancer cells. In patient tumor sections from high-stage, high-grade ovarian cancers, 7 to 10% of channels containing red blood cells were lined by tumor cells. By comparison, all vascular areas in benign tumors and low-stage cancers were endothelial lined. These results may offer new insights and molecular markers for consideration in ovarian cancer diagnosis and treatment strategies based on molecular vascular mimicry by aggressive tumor cells.     

Vasculogenic mimicry and tumor angiogenesis

Tumors require a blood supply for growth and hematogenous dissemination. Much attention has been focused on the role of angiogenesis-the recruitment of new vessels into a tumor from pre-existing vessels. However, angiogenesis may not be the only mechanism by which tumors acquire a microcirculation. Highly aggressive and metastatic melanoma cells are capable of forming highly patterned vascular channels in vitro that are composed of a basement membrane that stains positive with the periodic acid-Schiff (PAS) reagent in the absence of endothelial cells and fibroblasts. These channels formed in vitro are identical morphologically to PAS-positive channels in histological preparations from highly aggressive primary uveal melanomas, in the vertical growth phase of cutaneous melanomas, and in metastatic uveal and cutaneous melanoma. The generation of microvascular channels by genetically deregulated, aggressive tumor cells was termed "vasculogenic mimicry" to emphasize their de novo generation without participation by endothelial cells and independent of angiogenesis. Techniques designed to identify the tumor microcirculation by the staining of endothelial cells may not be applicable to tumors that express vasculogenic mimicry. Although it is not known if therapeutic strategies targeting endothelial cells will be effective in tumors whose blood supply is formed by tumor cells in the absence of angiogenesis, the biomechanical and molecular events that regulate vasculogenic mimicry provide opportunities for the development of novel forms of tumor-targeted treatments. The unique patterning characteristic of vasculogenic mimicry provides an opportunity to design noninvasive imaging techniques to detect highly aggressive neoplasms and their metastases.     

CD133和CD34在肝细胞癌血管生成拟态形成中的表达及意义

背景：在恶性肿瘤中血管生成拟态的形成过程与肿瘤干细胞有密切联系。 目的：分析肝癌干细胞标志物CD133和CD34在肝细胞癌血管生成拟态形成中的表达及意义。 方法：建立肝癌细胞HCC97H、SMMC7721和正常肝细胞L02三维培养体系,结合激光捕获显微切割技术分离形成血管生成拟态的肝癌细胞,分别利用RT-PCR和Western blot技术检测CD133和CD34表达水平。 结果与结论：三维培养条件下,肝癌细胞HCC97H细胞形成血管生成拟态,肝癌细胞SMMC7721以及正常肝细胞L02未形成血管生成拟态。形成血管生成拟态的肝癌细胞HCC97H中CD133、CD34在mRNA及蛋白表达水平上均高于未形成血管生成拟态的肝癌细胞SMMC7721和正常肝细胞L02(P < 0.05)。表明高侵袭性肝癌细胞在三维培养下形成血管生成拟态,而低侵袭性肝癌细胞及正常肝细胞不能形成血管生成拟态;肝癌细胞形成血管生成拟态的过程中与表达肝癌干细胞有关。     

Lipid droplets may lay a spacial foundation for vasculogenic mimicry formation in hepatocellular carcinoma

Vasculogenic mimicry is a highly patterned vascular channel distinguished from the endothelium-dependent blood vessel. Vasculogenic mimicry is lined by highly aggressive tumor cells, and is associated with tumor grade, invasion and metastasis, and poor clinical prognosis. Much attention has been focused on the signaling pathways and the tumor microenvironment needed for vasculogenic mimicry formation, however, the studies on the spacial foundation for vasculogenic mimicry formation are limited. There are many lipid droplets in hepatocellular carcinoma due to steatosis, while increased numbers of lipid droplets also have been reported in many other neoplastic processes. The role of lipid droplets in tumor is still unclear. Based on the similar structural and morphological characteristics between vasculogenic mimicry and lipid droplet, we speculate that the lipid droplets may lay a spacial foundation for vasculogenic mimicry formation by a way of “space placeholder” in HCC. Experimental data and limited clinical literatures support the hypothesis to a certain degree. This hypothesis may provide a new idea for the study of vasculogenic mimicry and also provide a new direction for the functional study of lipid droplets in tumor.     

Targeting Cancer Stem Cells to Modulate Alternative Vascularization Mechanisms

Recently, many papers have shown that tumor vascularization can be explained by angiogenesis, recruitment, cooption, vasculogenic mimicry and by mosaic vessels. In particular, vasculogenic mimicry seems to be different from mosaic blood vessels, where tumor cells form a part of the surface of the vessel while the remaining part is covered by endothelium. In this case, tumor cells in apparent contact with the lumen do not show an endothelial phenotype. More recently, vasculogenic mimicry was proposed to occur in patients with multiple myeloma due to bone marrow macrophages. Herein, all these data are, for the first time, discussed critically in comparison to cancer stem cells-which show high trans-differentiative capacity-and bone-marrow derived stem cells. In fact, the presence of alternative vasculogenic patterns might be due to the presence of stem cell population (cancer stem cells or bone-marrow stem cells). In this connection, the literature is discussed extensively and possible models are proposed. Pharmacological perspectives will also discuss.     

Vasodilator-stimulated phosphoprotein expression and its cytokine-mediated regulation in vasculogenesis during human placental development

Vasculogenesis and the subsequent step, angiogenesis, are the most important stages for the continuity of placental development. Vasodilator-stimulated phosphoprotein (VASP) has a widespread role in the control of cell motility and participates in filamentous actin formation. We hypothesized that VASP participates in vasculogenesis and angiogenesis, by regulating endothelial cell migration. We therefore studied VASP expression in vasculogenic sites in placenta throughout pregnancy and the effect of vascular endothelial growth factor (VEGF) and interleukin (IL)-8 on the regulation of VASP expression in placental explant cultures. We found that VASP is expressed in a spatially and temporally regulated manner by various cells of the villi. In the villous stroma, the most intense immunoreactivity was observed in vasculogenic areas and in endothelial cells. In the second and third trimesters, endothelial cells demonstrated weaker immunoreactivity for VASP compared to samples from first trimester. Ultrastructural analysis of corresponding sites for VASP showed that this protein was increased in pre-endothelial cells. Areas of the strongest VEGF and IL-8 expression by villous trophoblasts corresponded to the areas of strongest VASP expression by endothelial cells, and VEGF and IL-8 showed a stimulatory effect on VASP expression in placental explants (P < 0.05). These results suggest that VASP may participate in vasculogenesis and endothelial sprouting during placental vasculogenesis. In addition, one of the effects of VEGF and IL-8 in angiogenesis may be to induce VASP expression in a paracrine manner.