Basement membrane and the SIKVAV laminin-derived peptide promote tumor growth and metastases

Summary Laminin, the major glycoprotein component of basement membrane, promotes the malignant phenotype. Cells which are adherent to laminin are more malignant than the non-adherent cells and in certain tumor cells, the number of laminin receptors is positively correlated with malignancy. Laminin also increases collagenase IV activity, an enzyme demonstrated to be critical for tumor spread. A site on laminin, containing the amino acid sequence SIKVAV, has been identified which when injected intravenously with B16F10 melanoma cells, causes an increase in the number of colonies on the surface of the lungs. This peptide does not affect tumor cell arrest in the vasculature or the immune system. It does promote angiogenesis in various in vitro and in vivo models, thereby facilitating tumor cell survival.When a complex mixture of laminin-enriched basement membrane components (Matrigel) is coinjected with tumor cells subcutaneously, tumor incidence and growth increases. Various tumor cell lines and primary isolates, which previously could not form tumors in mice, can be induced to grow rapidly in the presence of Matrigel. Slowly growing tumors or arrested tumors can also be induced to grow more quickly with additional injections of Matrigel. When an SIKVAV-containing synthetic peptide is coinjected with B16F10 tumor cells and Matrigel subcutaneously in mice, larger tumors are formed than that observed with either Matrigel or cells alone. Such studies define the role of laminin in tumor growth and spread and generate new models for studying therapeutic agents. Of particular interest is the ability to grow primary isolates which generally do not grow in mice.     

The role of cell adhesion molecules in cancer invasion and metastasis

Summary Invasion and metastasis of tumor cells is the primary cause for the fatal outcome of cancer diseases. A striking feature of metastatic cells is the considerable flexibility in their adhesive interactions with other cells or components of the extracellular matrix. This review will describe the involvement of specific cell adhesion receptors, extracellular matrix molecules, and cell dissociating cytokines in the metastatic cascade. We will particularly focus on disturbance of intercellular adhesion as a prerequisite for the release of invasive cells from carcinomas. We suggest that cell dissociation in these tumors is accomplished by loss of function or expression of the epithelial cell adhesion molecule E-cadherin, and through the activity of cell motility factors, like scatter factor.     

The 67 kDa laminin receptor as a prognostic factor in human cancer

Different receptors for adhesion molecules, including the monomeric 67 kDa laminin receptor (67LR), are responsible for the interactions between tumor cells and components of the extracellular matrix that play an important role in tumor invasion and metastasis. Clinical data clearly demonstrate the importance of the 67LR in the progression of a wide variety of tumors, including breast, lung, ovary, and prostate carcinomas and lymphomas. Indeed, data on more than 4000 cases of different tumors from different organs studied by immunohistochemistry are all concordant with a role for the 67LR in invasiveness, metastasis, and even tumor growth. This receptor molecule appears to be unusual since the corresponding full-length gene encodes a 37 kDa precursor protein which, after acylation, dimerizes to generate the mature 67 kDa form. The primary function of the membrane receptor is to stabilize the binding of laminin to cell surface integrins, acting as an integrin-accessory molecule, although homology of the gene encoding the receptor precursor with other genes suggests additional functions. Studies conducted to define the structure, expression, and function of this laminin receptor represent a step toward developing therapeutic strategies that target this molecule. In particular, therapeutic approaches that downregulate expression of the receptor on tumor cells might lead to decreased tumor aggressiveness.     

Fibronectin in cell adhesion and invasion

Summary Fibronectin plays a major role in the adhesion of many cell types. The extent of cell adhesion in vitro is related not only to the ability of the cells to interact with matrix-bound fibronectin, when it is present, but also to the synthesis or lack of synthesis of fibronectin by the cells, and to the lack of deposition of synthesized fibronectin into an insoluble matrix surrounding the cells. Many malignant cells, regardless of whether they synthesize subnormal or normal amounts of fibronectin, fail to deposit that fibronectin into a surrounding insoluble matrix. The lack of fibronectin around such cells appears to reflect a general absence of extracellular matrix since other matrix components, such as collagen, laminin, and heparan sulfate proteoglycan, are concomitantly missing. Cells that lack their own cell surface fibronectin due either to lack of deposition or to lack of synthesis can nevertheless adhere to insoluble fibronectin matrices elaborated by other cells. These cellular characteristics appear to be associated with cell migration in vivo during embryogenesis, and the same characteristics may enhance the invasive potential of malignant cells. The remarkable effects that fibronectin has on cellular adhesion and the association of lack of extracellular matrix components with poorly differentiated and highly metastatic tumors in vivo mandates that more be learned about the molecular and cellular details of the interactions of cells with their surrounding matrix. Important information concerning tumor invasion will parallel such an understanding and may eventually become the basis for therapeutic approaches.This work was supported by grant CA 28896 and Cancer Center Support Grant CA 30199 from the National Cancer Institute, Department of Health and Human Services.     

Molecular and cellular analysis of basement membrane invasion by human breast cancer cells in Matrigel-basedin vitro assays

Summary In vitro analyses of basement membrane invasiveness employing Matrigel (a murine tumor extract rich in basement membrane components) have been performed on human breast cancer model systems. Constitutive invasiveness of different human breast cancer (HBC) cell lines has been examined as well as regulation by steroid hormones, growth factors, and oncogenes. Carcinoma cells exhibiting a mesenchymal-like phenotype (vimentin expression, lack of cell border associated uvomorulin) show dramatically increased motility, invasiveness, and metastatic potential in nude mice. These findings support the hypothesis that epithelial to mesenchymal transition (EMT)-like events may be instrumental in the metastatic progression of human breast cancer. The MCF-7 subline MCF-7_ADR appears to have undergone such a transition. The importance of such a transition may be reflected in the emergence of vimentin expression as an indicator of poor prognosis in HBC. Matrix degradation and laminin recognition are highlighted as potential targets for antimetastatic therapy, and analyses of laminin attachment and the matrix metalloproteinase (MMP) family in HBC cell lines are summarized. Matrigel-based assays have proved useful in the study of the molecular mechanisms of basement membrane invasiveness, their regulation in HBC cells, and their potential as targets for antimetastatic therapy.     

The urokinase-type plasminogen activator system in prostate cancer metastasis

Accumulated clinical and experimental evidence indicates that the urokinase-type plasminogen activator (uPA) and its regulators are causatively involved in the metastatic phenotype of many types of cancers. In the past couple of decades, investigation on the role of the uPA system in human prostate cancer (PC) has been intensified and has yielded valuable insights. This review summarizes recent advances made in several areas regarding the clinical relevance, the function and the molecular mechanisms of the uPA system in PC metastasis. A current consensus suggests that the uPA system promotes PC metastasis by mediating pericellular plasminogen activation. Towards the development of therapeutic strategies that specifically target uPA-mediated PC metastasis, several remaining issues are discussed.     

Glycosidases in cancer and invasion

Summary Glycosidases have been demonstrated to be elevated in the interstitial fluid of tumors, sera of animals and patients with tumors, and in some tumor tissue as compared to normal adjacent tissue. Elevations of serum -N-acetylglucosaminidase and -glucuronidase most commonly have been found to occur and these enzymes have been shown to be secreted into the extracellular medium by many different tumor cell types in vitro. The mechanism of cellular release of these hydrolytic enzymes probably involves tumor lysosomal exocytosis. Increased tumor glycosidase levels may promote increased tumor cell shedding from primary tumors, local invasion and perhaps be responsible directly, or indirectly for structural changes in tumor cell surface glycoconjugates. These cell surface changes could facilitate tumor cell thrombus formation, secondary site implantation and attachment in the microcirculation to endothelial cells and/or subendothelial basement membrane components.Other studies have demonstrated a correlation between metastatic cell potential and increased endoglycosidase and polysaccharide lyase activity. Generally, metastatic tumor cell variants have been found to be more invasive and capable of degrading proteoglycan basement membrane components, in part due to these increased levels of degradative enzymes. Hence, it is of considerable interest to develop inhibitors against these enzymes. Initial studies with glucuronidase inhibitors in the therapy of bladder tumors have been promising and with the advent of better agents and the use of appropriate in vitro metastatic models it may be possible to design and develop agents which interfere in various metastatic events and limit tumor progression.Supported in part by grants from the National Cancer Institute, CA-13038 and CA-08963.     

Contributions of tumor and stromal matrix metalloproteinases to tumor progression,invasion and metastasis

Summary The malignant progression of tumors is driven by the expression of oncogenes and loss of expression of tumor suppressor genes; factors that are intrinsic to cancer cells. The phenotypic changes brought about by the gain or loss of expression of oncogenes and tumor suppressor genes lead to the acquisition of malignant traits, namely, the ability to invade into and grow in ectopic tissue environments. Recently, however, focus in cancer research has widened from the cancer cell to include the surrounding tumor stroma as an integral player in the process of tumor progression. One of the areas in cancer research contributing to this enhanced appreciation of stromal involvement in tumor progression and metastasis is that of matrix metalloproteinases (MMPs).This review provides an overview of the characteristics of MMPs and discusses their role in the progression and metastasis of tumors. Initially, attention will focus on the regulation of MMPs in tumor cells but will switch to discourse on stromal expression of MMPs in tumors and speculation on the functional consequences of stromal expression of MMPs.     

Skp2在恶性肿瘤侵袭转移中的作用

SCF(Skpl-Cullinl-F-box)复合物是一大类泛素连接酶E3.S期激酶相关蛋白2(S-phase kinase associated protein 2,Skp2)是F盒蛋白家庭成员之一.研究表明Skp2主要参与细胞周期的调控,与多种恶性肿瘤的发生、发展密切相关.近年研究发现,Skp2在肿瘤的侵袭、转移中也扮演着重要的角色.Skp2可以通过调控RhoA、基质金属蛋白酶MMP-2、MMP-9的表达和活性以及E-cadherin的降解从而促进肿瘤细胞的侵袭和转移.本文就近几年Skp2在肿瘤侵袭、转移中的作用进行综述.     

Malignant melanoma metastasis to brain: role of degradative enzymes and responses to paracrine growth factors

Mouse and human melanoma cells metastatic to the brain express degradative enzyme activities that are used for invasion of brain basement membrane and parenchyma. Compared to poorly metastatic or lung- or ovary-metastatic murine melanoma lines, the brain-metastatic sublines secreted higher levels of a variety of degradative enzymes. Brain-metastatic murine and human melanoma cells also degraded subendothelial basement membrane and reconstituted basement membrane at rates higher than other metastatic melanoma cells. In some cases these degradative activities in mouse and human melanoma cells can be induced by paracrine factors known to be present in the brain parenchyma, such as nerve growth factor (NGF). NGF stimulates the expression of degradative enzymes, such as the endo--glucuronidase heparanase, that are important in basement membrane penetration but this factor does not stimulate melanoma cell growth. The growth of brain-metastasizing melanoma cells appears to be stimulated by other paracrine growth factors, such as paracrine transferrin. Melanoma cells metastatic to brain express higher numbers of transferrin receptors and respond and proliferate at lower concentrations of transferrin than do melanoma cells metastatic to other sites or poorly metastatic melanoma cells. The results suggest that degradation and invasion of brain basement membrane and responses to paracrine neurotrophins and paracrine transferrins are important properties in brain metastasis of murine and human malignant melanoma cells.     

Transgenic mouse models of breast cancer

Summary Although valuable initial information can be gathered about transformation fromin vitro studies, human cancer occurs in the context of a complex interaction with its environment and must ultimately be studied in living animals. Transgenic animal models have been used to study breast transformation for a number of years and have yielded valuable information on the subject. In this paper, we will summarize results from our laboratories, and others, regarding the use of transgenic mice to study breast tumorigenesis. We will also suggest future directions for the use of transgenic models to understand, and hopefully, one day to cure the disease. Note: genes are referred to as lowercase names in italics (e.g.myc) and their protein products as uppercase (e.g. Myc).     

FBXO31 promotes cell proliferation,metastasis and invasion in lung cancer

FBXO31 is a member of F-box family which is involved in diverse biological functions and development of disease. Recent reports in breast cancer, hepatocellular carcinoma and ovarian cancer demonstrated inhibitory effect of FBXO31 on proliferation and tumorigenesis. However, the function of FBXO31 is not analyzed in lung cancer so far. In this study, we reported that expression of FBXO31 was higher in lung cancer tissues compared with non-cancerous lung tissues, and that higher expression of FBXO31 was significantly associated with tumor size, tumor infiltration, clinical stages and lymph node metastasis. In addition, exogenous expression of FBXO31 promoted cell growth, metastasis and invasion in A549 cells. Conversely, silencing FBXO31 by specific siRNA caused inhibitory effect on cell growth, metastasis and invasion. Moreover, tumorigenicity assays in nude mice showed FBXO31 promoted tumor growth in vivo. In conclusion, our data suggest FBXO31 promotes cell proliferation, metastasis and invasion in lung cancer.     

Distribution of Ha-RAS-1 proto-oncogene alleles in breast cancer patients and in a control population

Summary The frequencies of 13 different Ha-ras proto-oncogene alleles have been estimated in 92 breast cancer patients and 60 unaffected individuals. The Ha-ras alleles can be identified using a DNA restriction fragment length polymorphism (RFLP) closely linked to the 3 end of the gene, and are characterized by a different length due to a region of sequences repeated a variable number of times (variable tandem repeats, VTR).The statistical analysis of the data obtained shows that the frequency of alleles ranging between specific length limits is significantly higher in breast cancer patients than in controls. The same applies to specific genotypes bearing the aforementioned alleles. This suggests that the inheritance of these alleles may be associated with an increased risk of developing breast cancer.     

P2Y2 receptor promotes cell invasion and metastasis in prostate cancer cells

Background:

Our previous study demonstrated that extracellular adenosine 5′-triphosphate (ATP) stimulated prostate cancer cell invasion via P2Y receptors. However, the purinergic receptor subtype(s) involved in this process remains unclear. Here we aimed to determine whether P2Y2, one subtype of P2Y receptors, was involved in the invasion and metastasis of prostate cancer cells, and elucidated the underlying mechanism.

Methods:

RNAi was introduced to silence the expression of P2Y2. In vitro invasion and migration assays and in vivo experiments were carried out to examine the role of P2Y2 receptor in cell invasion and metastasis. cDNA microarray was performed to identify the differentially expressed genes downstream of ATP treatment.

Results:

P2Y2 was significantly expressed in the prostate cancer cells. Knockdown of P2Y2 receptor suppressed cell invasion and metastasis in vitro and in vivo. Further experiments identified that ATP could promote IL-8 and Snail expression and inhibit E-cadherin and Claudin-1 expression. Knockdown of P2Y2 receptor affected the expression of these EMT/invasion-related genes in vitro and in vivo.

Conclusion:

P2Y2 receptor promotes cell invasion and metastasis in prostate cancer cells via some EMT/invasion-related genes. Thereby, P2Y2 receptor could be a potential therapeutic target for the treatment of prostate cancer.     

The role of growth factors in breast cancer

A series of teleconferences has been organized under the auspices of Bristol-Myers to address several major current questions in oncology. A panel of recognized experts with a moderator has been assembled to discuss each question, and we are reporting a number of these discussions in Breast Cancer Research and Treatment. This is reprinted from Oncology Viewpoints, courtesy of Bristol-Myers Oncology Division, Evansville IN 47721, USA.     

CXCL12-CXCR4在胰腺癌侵袭转移中的作用

目前研究发现趋化因子及受体CXCL12-CXCR4生物轴在胰腺癌的侵袭转移中起重要作用.CXCL12-CXCR4可通过促进胰腺癌细胞增殖、增强癌细胞的迁移、促进细胞外基质蛋白降解和血管新生等机制而促进胰腺癌的侵袭转移.     

MIG-7 and phosphorylated prohibitin coordinately regulate lung cancer invasion/metastasis

Growth factors and COX-2/PGE2 enhance lung cancer invasion/metastasis via PI3K/Akt and RAS/Raf. Here, we explored their mechanism of action further. We found first that higher levels of migration inducting gene-7 protein (MIG-7) and PHB phosphorylated at threonine 258 (phospho-PHB^T258) are positively correlated with advanced stages of human lung cancer in tissue microarray. PGE2 or growth factors such as EGF, HGF and IGF-1 increased complex formation of phospho-PHB^T258 with Ras, phospho-Akt^S473, phospho-Raf-1^S338, MEKK1 and IKKα/β^S176/180 in the raft domain transiently within 1 hour and MIG-7 in the cytosol 12-24 hours later. Association of phospho-PHB^T258 with MEKK1 but not MEKK3 activates IKK/IκB/NF-κB and MEK/ERK to increase cellular COX-2/PGE2 and an E-cadherin suppressor Snail leading to enhancement of epithelial-mesenchymal transition (EMT) and lung cancer migration/invasion. MIG-7, on the other hand, was induced by growth factors and PGE2 via Akt/GSK-3β in a phospho-PHB^T258 independent manner. MIG-7 increased two E-cadherin suppressors ZEB-1 and Twist to enhance EMT and cancer migration/invasion. Downregulating phospho-PHB^T258 and MIG-7 had an additive effect on attenuating lung cancer invasion/metastasis and prolonging the survival of xenograft mice. Phospho-PHB^T258 and MIG-7 may thus play complementary roles in the initiation and sustainment of the effects of growth factors and COX-2/PGE2 on cancer invasion/metastasis.     

Inhibition of invasion and metastasis by glypican-3 in a syngeneic breast cancer model

Glypican-3 (GPC3), a proteoglycan bound to the cell membrane through a GPI anchor, is widely expressed in the embryo but down regulated in most adult tissues, with some exceptions as mammary cells. GPC3 is involved in the regulation of cell proliferation and survival in specific cell types. LM3, a murine mammary tumor cell line unable to express GPC3, was stably transfected with the rat GPC3 gene to analyze its role in tumor progression. Upon injection into syngeneic BALB/c mice LM3-GPC3 clones showed less local invasiveness and developed fewer spontaneous and experimental lung metastasis than controls. GPC3-expressing cells were more sensitive to apoptosis induced by serum depletion, exhibited a delay in the first steps of spreading and were less motile than controls. On the other hand, LM3-GPC3 cells were significantly more adherent to FN than control ones. We observed that GPC3 transfectants presented a higher expression of E-cadherin and -catenin, molecules whose down regulation has been associated with tumor progression. Exogenous TGF- increased MMP-9 activity in both control and GPC3-expressing cells, but did not modulate MMP-2. Contrarily, GPC3 expression prevented the increase of MMP-2 activity induced by IGF-II. Our results suggest that GPC3 has a protective role against mammary cancer progression.     

谷氨酸在调控胃癌细胞侵袭转移中的作用

目的:探讨谷氨酸受体NR1亚基在胃癌中的表达及谷氨酸对胃癌侵袭转移的调控作用。方法:运用Real-time PCR方法检测人胃癌高低转移细胞系中NR1亚基的表达情况;体外侵袭实验检测谷氨酸对胃癌细胞侵袭能力的影响及N-甲基-D-天冬氨酸受体（NMDA）的特异性阻断剂对胃癌细胞侵袭能力的阻断作用;全细胞膜片钳技术检测胃癌细胞上的谷氨酸受体诱发电流。结果:人胃癌细胞可表达NR1亚基,且NR1亚基在胃癌高转移细胞系中的表达量高于其在低转移细胞系中的表达量（P<0.05）;谷氨酸可增强胃癌细胞的侵袭能力,而阻断剂MK801可降低胃癌细胞的侵袭能力（P<0.05）;谷氨酸刺激后,胃癌细胞可产生谷氨酸受体诱发电流,MK801可部分有效阻断谷氨酸受体诱发电流。结论:谷氨酸受体NR1亚基在胃癌细胞中表达明显升高,谷氨酸与其结合可促进胃癌侵袭转移的发生,而特异性受体阻断剂可部分阻断其作用。