Expression of CD44v6 in advanced gastric cancer and its relationship to hematogenous metastasis and long-term prognosis

BACKGROUND AND OBJECTIVES: Variants of CD44 have been proposed to be important in promoting tumor progression and metastasis. We attempted to determine the expression of CD44v6 product in advanced gastric cancer and to evaluate its prognostic value. METHODS: The expression of CD44v6 was analyzed immunohistochemically in advanced gastric cancers using monoclonal antibody, 44-2V. We investigated the relationship between CD44v6 expression and prognosis in 201 gastric cancer patients. RESULTS: Ninety-five (47.3%) of 201 cancer tissues expressed CD44v6. The expression of CD44v6 protein was significantly higher in differentiated, adenocarcinoma than in diffuse type carcinoma. The CD44v6-positive cancers were more frequently associated with hematogenous metastasis. There was no significant correlation between CD44v6 immunoreactivity, and prognosis among the combined cases. Among patients with differentiated adenocarcinoma, however, the prognosis was significantly poorer in patients with CD44v6-positive tumors than in those with CD44v6-negative tumors. CONCLUSIONS: CD44v6 protein may have an important role in hematogenous metastasis, and may be a biologic marker of prognostic significance in differentiated type gastric cancers.     

Radical treatment of brain metastasis of colorectal-cancer

Among 15 patients with brain metastases from colorectal cancer, the eight patients treated by open surgery+whole brain irradiation (35-45 Gy) or gamma-knife surgery had significant improvement in their neurologic symptoms and were able to return to their daily life. Survival from the time of diagnosis of brain metastases to death averaged 7.6 months in these radically-treated patients compared to 1.7 months in the conventional group (p<0.05). If performance status permits, aggressive treatment such as open surgery or gamma-knife surgery should be an option for treatment of brain metastasis of colorectal cancer.     

Prevention of hematogenous metastasis by neutralizing mice and its chimeric anti-Aggrus/podoplanin antibodies

The platelet aggregation-inducing factor, Aggrus (also known as podoplanin), is reported to contribute to cancer metastasis by mediating cancer cell-platelet interaction. Aggrus has been shown to be upregulated in many different types of cancers. Thus, not only the functional inhibition of Aggrus, but also its application as a cancer-specific antigen has therapeutic potential. Among a series of anti-Aggrus mAb established previously, no mouse anti-human Aggrus mAb exists that possesses the ability to neutralize platelet aggregation. For precise preclinical examinations of mouse and monkey models, the establishment of Aggrus-neutralizing mouse mAb and their chimeric Abs is needed. In this study, we established two mouse anti-human Aggrus mAb, P2-0 and HAG-3. A precise analysis of their epitopes revealed that P2-0 recognized the conformation near the bioactive O-glycosylation site at the Thr(52) residue. In contrast, HAG-3 recognized the amino-terminus side at a short distance from the conformation recognized by P2-0. We observed that only P2-0 attenuated Aggrus-induced platelet aggregation and Aggrus binding to its platelet receptor, that is, the C-type lectin-like receptor-2. Consistent with these data, only P2-0 prevented the experimental metastasis of human Aggrus-overexpressing CHO cells. Subsequently, we cloned the complementary determining region of P2-0 and produced the murine/human chimeric P2-0 antibody. This chimeric antibody maintained its inhibitory activity of Aggrus-induced platelet aggregation and experimental metastasis. Thus, P2-0 and its chimeric antibody are expected to aid the development of preclinical and clinical examinations of Aggrus-targeted cancer therapy.     

Soluble tumour-specific antigen and its relationship to tumour growth.

Hooded rats bearing a syngeneic methylcholanthrene-induced tumour were evaluated for extent of in vivo host immunity and this was correlated by in vitro techniques with the levels of circulating tumour antigen and specific antibody. Early tumour growth was associated with detectable immunity, as measured by the capacity of the animal to reject a second direct challenge of the same tumour at a remote site. Radioimmunoassay for circulating tumour antigen and indirect membrane immunofluorescence for antitumour antibody did not detect either component at this stage. Animals with advanced tumours lost immunity as detected by direct tumour challenge, and this closely coincided with the appearance of rising levels of circulating soluble tumour antigen. Although the host possessed the immunologic ability to react against its own neoplasm, this ability was insufficient to produce tumour rejection. Active immunotherapy initiated at the time of, or up to 10 days after, intramuscular challenge with tumour, increased tumour immunity sufficiently for tumour growth to be prevented. Successful immunization was associated with the early appearance (16 days) of measurable levels of antitumour antibody and absence of circulating antigen. It is concluded that soluble tumour antigen present in the local microenvironment of the tumour in the early stages of tumour growth interferes with the ability of immune cells to cause tumour rejection. As the tumour progressively grows, sufficient soluble antigen is produced and released systemically to suppress the effector arm of the host's tumour immune response at distant sites. The levels of circulating soluble tumour antigen attained may be of critical importance in the suppression of rejection responses that prevent metastasis.     

The metastatic T-cell hybridoma antigen/P-selectin glycoprotein ligand 1 is required for hematogenous metastasis of lymphomas

Using variants of the murine BW5147 lymphoma cell-line, we have previously identified 3 monoclonal antibodies (MAbs) that discriminate between metastatic and nonmetastatic BW5147-derived T-cell hybridomas and lymphomas, as well as BW5147-unrelated T-lymphomas. These MAbs were reported to recognize an identical membrane-associated sialoglycoprotein, termed "metastatic T-cell hybridoma antigen" (MTH-Ag). Here, we document that the expression pattern of the MTH-Ag on metastatic and nonmetastatic BW5147 variants correlates with that of the P-selectin glycoprotein ligand 1 (PSGL-1), a sialomucin involved in leukocyte recruitment to sites of inflammation. Moreover, the MAbs against the MTH-Ag recognize PSGL-1 when it is transfected in MTH-Ag-negative BW5147 variants, suggesting that the MTH-Ag is PSGL-1. Overexpression of MTH-Ag/PSGL-1 in MTH-Ag-negative BW5147 variants did not affect their in vivo malignancy. Yet, down-regulation of MTH-Ag/PSGL-1 expression on metastatic, MTH-Ag-positive BW5147 variants, using an RNA interference (RNAi) approach, resulted, in a dose-dependent manner, in a significant reduction of liver and spleen colonization and a delay in mortality of the recipient mice upon intravenous inoculation. Collectively, these results demonstrate that, although MTH-Ag/PSGL-1 overexpression alone may not be sufficient for successful dissemination and organ colonization, MTH-Ag/PSGL-1 plays a critical role in hematogenous metastasis of lymphoid cancer cells.     

Endothelial Rac1 is essential for hematogenous metastasis to the lung

A variety of vasoactive stimuli induce endothelial permeability through Rac1, a membrane of Rho small GTPases. Here, we determine whether tumor-secreted vasoactive stimulant through Rac1 inducing permeability contributes to hematogenous metastasis. Activation of Rac1 was assayed in human umbilical vein endothelial cells (HUVEC), transendothelial passages were measured by Transwell chambers, and hematogenously metastatic mouse model was generated by intravenous injection with Lewis lung carcinoma cells (LLC). LLC secreted abundant vascular endothelial growth factor (VEGF) in the culture media and sera of mice bearing LLC xenografts or metastatic LLC, and VEGF activated Rac1 through VEGF receptors/PI3Kβ signaling cascade, resulting in hyperoxidative stress and consequent hyperpermeability in HUVEC. Moreover, in co-culture of LLC and HUVEC, significant increases in endothelial permeability and transendothelial migration of LLC were robustly attenuated by either anti-VEGF neutralizing antibody or Rac1 knockdown in HUVEC. Finally, in metastatic mouse model, deletion of one copy of Rac1 in endothelium not only significantly attenuated LLC-induced vascular permeability, but robustly reduced the metastasis of LLC to lungs. This study supports that tumor-secreted vasoactive stimuli activate Rac1 to induce permeability and consequent transendothelial migration of tumor cells, and that loss of Rac1 function in endothelium is an effective therapeutic intervention for hematogenous metastasis.     

Detection of hematogenous bone metastasis in cervical cancer

BACKGROUND:

In this large‐scale, retrospective study, the authors evaluated the diagnostic performances of computed tomography (CT), magnetic resonance (MR) imaging, and ¹⁸F‐fluorodeoxyglucose‐positron emission tomography (¹⁸F‐FDG–PET) in detecting hematogenous bone metastasis in patients with cervical cancer. The associated risk factors also were analyzed.

METHODS:

Patients with invasive cervical cancer who had both ¹⁸F‐FDG–PET studies and CT or MR imaging studies were selected. Patients who had either International Federation of Gynecology and Obstetrics (FIGO) stage III/IV disease or positive lymph node metastasis at the time of primary staging and patients who had suspected recurrent disease were included in the analyses. The diagnostic performances of PET was compared with the performance of CT and MR imaging by using the area under the receiver‐operating‐characteristic curve (AUC). Both univariate and multivariate analyses were applied to assess the risk factors for hematogenous bone metastasis at primary staging.

RESULTS:

PET was more sensitive than CT (P = .004) and was more specific than MR imaging (P = .04). The diagnostic performance of PET was significantly superior to the performance CT (AUC, 0.964 vs 0.662; P < .001) and MR (AUC, 0.966 vs 0.833; P = .033). Both FIGO stage and the extent of lymph node metastases were associated with hematogenous bone metastasis in univariate analysis. However, the extent of lymph node metastases was the only significant risk factor in multivariate analysis (P = .025).

CONCLUSIONS:

The current study demonstrated the superiority of ¹⁸F‐FDG–PET over CT and MR imaging for detecting hematogenous bone metastasis in patients with advanced cervical cancer. Hematogenous bone metastasis in cervical cancer was associated with the extent of lymph node metastases rather than with FIGO stage. Cancer 2009. © 2009 American Cancer Society.     

Circulating levels of epithelial membrane antigen in patients with breast or colorectal-cancer

Epithelial membrane antigen (EMA) is expressed by adenocarcinomas of the breast, ovary and colon and has been suggested as a circulating tumour marker. Serum EMA levels were measured in 126 patients, 31 with colorectal cancer, 52 with breast cancer and 43 age matched controls using a competitive binding radioimmunoassay and the rat anti-EMA monoclonal antibody (MAb) ICR2. The EMA levels in the control group varied widely from 90-3240ng/ml with a median value of 570ng/ml. This was not significantly different from the levels in patients with colorectal cancer (60-8530, median 580ng/ml) or those with breast cancer (210-13300, median 655ng/ml). However, the highest EMA levels (>5000ng/ml) were found in patients with cancer. The wide range of EMA levels in the control group prohibit its use for screening.     

Robo1蛋白在乳腺癌中的表达及其与乳腺癌脑转移的关系

目的 检测Robo1蛋白在不同乳腺肿瘤组织中的表达情况,并探讨Robo1蛋白的表达与乳腺癌脑转移的关系.方法 采用链霉素抗生物素蛋白-生物素(LSAB)法,对24例发生脑转移的乳腺浸润性导管癌、71例未发生脑转移的乳腺浸润性导管癌、22例乳腺导管内癌和23例乳腺纤维腺瘤组织中Robo1蛋白的表达情况进行检测.结果 Robo1蛋白在导管内癌和浸润性导管癌组织中的阳性表达率分别为59.1%和45.3%,均明显低于其在纤维腺瘤组织中的阳性表达率(87.0%,P＜0.05).Robo1蛋白在有脑转移的浸润性导管癌组织中的阳性表达率为12.5%,明显低于其在无脑转移的浸润性导管癌中的阳性表达率(56.3%,P＜0.05).Robo1蛋白在＞50岁乳腺浸润性导管癌患者中的阳性表达率为57.8%,明显高于其在≤50岁乳腺浸润性导管癌患者中的阳性表达率(34.0%,P＜0.05).Robo1蛋白阴性表达患者的5年生存率为57.7%,明显低于阳性表达的患者(83.7%,P<0.05).Robo1蛋白在乳腺浸润性导管癌中的表达与肿瘤大小、淋巴结转移状态、病理学分期、组织学分级以及临床分期均无关(均P＞0.05).结论 Robo1蛋白在乳腺浸润性导管癌中的表达与脑转移呈负相关,与患者的发病年龄及预后呈正相关,可成为判断乳腺癌预后和脑转移的分子标志物.

Abstract：

Objective To detect the expression of Robo1 in different breast tumors and its association with the breast cancer brain metastasis. Methods Labelled streptavidin-biotin (LSAB) staining was used to examine the Robo1 expression in specimens from 24 cases of invasive ductal carcinoma(IDC)with brain metastasis, 71 cases of IDC without brain metastasis, 22 cases of ductal carcinoma in situ (DCIS) and 23 cases of fibroadenoma. Results The expression pattern of Robo1 in DCIS (59.1%) and IDC (45.3%) was significantly lower than that in adenofibroma (87.0%, P<0.05). The expression of Robo1 in IDC with brain metastasis (12.5%) was significantly lower than that in IDC without brain metastasis (56.3%, P<0.05). The expression of Robo1 was much higher in more than 50 year-old-group (57.8%) than that in less than 50 year-old-group (34.0%) of IDC patients. The overall survival time in patients with the Robo1 negative expression was significantly shorter than those with positive expression (P<0.05). No correlation was found between the Robo1 expression and the tumor size, lymph node metastasis, pathologic stage, histological grade and clinical stage (P>0.05). Conclusions The Robo1 expression correlates negatively with IDC brain metastasis, and correlates positively with the age and prognosis of IDC patients. Robo1 may be applied as a marker in evaluation of the IDC prognosis and brain metastasis.     

Expression of tissue factor in non-small-cell lung cancers and its relationship to metastasis

Tissue factor (TF) is an initiator of the extrinsic cascade of blood coagulation. Although recent studies have revealed a relationship between metastatic properties and TF expression in some neoplastic cells, the significance of TF in lung cancer, especially in non-small-cell lung cancer (NSCLC), is still unclear. In this study, TF was detected in NSCLC cell lines by functional study, Western blot analysis and immunocytochemical staining. TF levels in eight NSCLC cell lines were also quantitated by enzyme-linked immunosorbent assay (ELISA), and TF expression was evaluated in 55 specimens of surgically resected NSCLCs. NSCLC cell lines derived from metastatic lesions produced high levels of TF (48.3+/-23.5 ng 10(-6) cells, mean +/- s.e.m.), whereas those derived from primary lesions produced low levels of TF (0.2+/-0.1 ng 10(-6) cells). Immunohistochemical studies disclosed significantly stronger staining for TF in cells from NSCLC patients with metastasis than in those without metastasis. Among the 28 patients with metastasis, ten were strongly positive, 16 were moderately positive and two were negative for TF. In contrast, among the 27 patients without metastasis, only two were strongly positive, 18 were moderately positive and seven were negative for TF. Therefore, malignant cells from patients with lung cancer produce various levels of TF, and TF may play an important role in the metastatic process.     

Contributions of vascularized lymph-node metastases to hematogenous metastasis in a rat mammary carcinoma

Rats received hind-foot-web (FWI) injections of MT-100-TC mammary carcinoma cells; the resultant tumor metastasized first to the popliteal lymph nodes. Over the course of 4 weeks, in association with increases in tumor weight, the blood-flow to the popliteal nodes increased 18-fold, and their vascular densities increased 2-fold. In spite of this vascularization, cancer cells were detected in only 3 of 648 blood vessels associated with involved, ipsilateral lymph nodes compared with intravascular cells in 82 of 314 vessels associated with "primary" foot-pad lesions. The presence of tumorigenic cancer cells in the right ventricular blood of animals bearing these tumors is, therefore, considered to result from their direct entry into blood vessels from the "primary" lesions, and/or from extra-nodal invasion of vessels in tissues to which nodal tumors were adherent, as distinct from passage via lymphatico-venous communications between tumors and nodal blood-vessels. The reconstructed events occurring in the rat model, with effective restriction of regional node metastases to the nodes themselves for a time, could possibly account for the long-term survival of some patients with breast cancer and regional-node metastases, following surgery.     

Lymphogenous and hematogenous metastasis of Lewis lung carcinoma in the mouse

Quantitative and temporal comparisons were made between lymphogenous and hematogenous metastasis in the non-immunogenic model system of mice bearing transplanted 3LL tumors in their hind feet. The experimental evidence indicates that cancer cells disseminate from clinically detectable primary cancers by non-exclusive routes in the blood-stream and in the lymphatics. Following a delay of approximately 2 weeks after injection of 3LL cancer cells into the foot, local lymph-node micrometastases occur, together with the first appearance of overt hematogenous metastases in the lungs. The anatomic extent of lymph-node involvement, determined by bioassays of orthotopic grafts, of ipsilateral popliteal, inguinal and lumbar nodes, provides an accurate indicator of hematogenous metastasis, even though lymphogenous and hematogenous metastasis are operationally independent in this tumor/host system.     

Serum sialyl-Tn antigen in colorectal cancer and its relationship to prognosis]

Serum sialyl-Tn antigen level was studied in 117 cases of colorectal cancer. The positive rates for serum sialyl-Tn antigen were 23.1% in primary colorectal cancer. There is a significant correlation between the positive rates of serum sialyl-Tn antigen and liver metastases, peritoneal dissemination, or lymph node metastases. The survival rate for patients with sialyl-Tn antigen positive sera was lower than those with sialyl-Tn antigen negative. Among patients who underwent curative operation, patients with sialyl-Tn positive sera gave a recurrence rate of 35.7%, while those with sialyl-Tn negative sera recorded a recurrence rate of 14.7%. There is no correlation between the positive rates of serum sialyl-Tn antigen and DNA ploidy pattern. Serum sialyl-Tn antigen levels may be a useful indicator in projecting the prognosis of colorectal cancer.     

Effects of DN-9693, a new metastasis inhibitor, on murine hematogenous metastasis

1,5-Dihydro-7-(1-piperidinyl)-imidazo[2,1-b]quinazolin-2(3H)-one dihydrochloride hydrate (DN-9693), a new c-AMP: phosphodiesterase inhibitor was examined for its inhibitory effects on platelet aggregation induced by metastasizing tumor cells and on blood-borne metastases of these tumors. 1-3 microM of DN-9693 completely inhibited platelet aggregation induced by B16 melanoma subline BL6 and Lewis lung carcinoma (3LL) cells. Platelets prepared from mice intravenously or orally administered with DN-9693 failed to aggregate after the addition of BL6 cells. Intravenous injection of DN-9693 was effective in protecting the mice inoculated with 1 X 10(6) BL6 cells against acute pulmonary embolic death. Either intravenous or oral administration of DN-9693 (1-10 mg/kg) sufficiently suppressed thrombus formation and subsequent pulmonary metastasis caused by intravenously inoculated BL6 or 3LL cells. Spontaneous pulmonary metastasis of 3LL was also inhibited by DN-9693. Continuous administration of DN-9693 during and after surgical excision of the primary tumors was the most effective treatment against the development of pulmonary metastases of 3LL.     

乳腺癌组织CXCR7表达与淋巴结转移相关性的探讨

目的:探讨趋化因子受体7(CXCR7)表达与乳腺癌淋巴结转移的关系.方法:用组织芯片及免疫组化技术检测CXCR7在乳腺癌组织及正常组织中的表达.结果:CXCR7在淋巴结转移组中呈高表达(83.33％,40/48),显著高于无淋巴结转移组(61.11％,33/54),P＜0.05;CXCR7表达与乳腺癌淋巴结转移(P=0.013)、肿瘤大小(P=0.000)有关,与年龄(P=0.876)、分化程度(P=0.501)、ER(P=0.057)和PR(P=0.885)无关.结论:CXCR7高表达与乳腺癌淋巴结转移具有一定的相关性,选择性阻断CXCR7的作用,对控制乳腺癌淋巴结转移可能是一个有效的途径.     

Biomechanical interactions of cancer cells with the microvasculature during hematogenous metastasis

Different aspects of hematogenous metastasis are discussed from the viewpoint of biomechanics. The processes considered include the role of primary tumor pressure, cell locomotor forces and degradation, in invasion of tissues and intravasation by cancer cells. Consideration of the fluid dynamics of cancer cell movement along capillaries lead to the view thatin vivo, arrest is primarily due to mechanical trapping of cancer cells, and that the pathobiologic role of so-called adhesion molecules is not mainly in the arrest and adhesion of cancer cells, but rather in stimulating their proliferation by signal induction. As a consequence of deformation from spherical-to-cylindrical shape in the microvasculature, demands for increased surface membrane area leads to increases in surface membrane tension above critical levels for rupture, and the cancer cells are rapidly and lethally damaged. The possibility is briefly discussed of increasing the susceptibility of circulating cancer cells to mechanical trauma, as a form of anti-metastatic therapy.     

Occurrence of hematogenous metastasis and serum levels of thymidine phosphorylase in colorectal cancer

Thymidine phosphorylase (dThdPase) is known to promote the development of new blood vessels. Increased dThdPase expression in solid tumors has been shown to correlate with tumor growth, invasion and metastasis. In the present study, we measured dThdPase levels in the tumor tissue and in the serum from the tumor drainage and peripheral venous blood obtained from patients with resectable colorectal cancer. Serum dThdPase levels, measured by a modified ELISA method, were significantly higher in patients with hematogenous metastasis. In the tumor tissue specimens, no significant difference was observed between patients with or without hematogenous metastasis. These results suggest that the serum dThdPase levels are a novel marker to predict occurrence of hematogenous metastasis in patients with resectable colorectal cancer.