The focal adhesion-associated proteins DOCK5 and GIT2 comprise a rheostat in control of epithelial invasion.

DOCK proteins are guanine nucleotide exchange factors for Rac and Cdc42 GTPases. DOCK1 is the founding member of the family and acts downstream of integrins via the canonical Crk-p130Cas complex to activate Rac GTPases in numerous contexts. In contrast, DOCK5, which possesses the greatest similarity to DOCK1, remains sparingly studied. Here we establish that DOCK5 has a non-redundant role in regulating motile and invasive capacities of epithelial cells. DOCK1 is constitutively associated with sites of integrin attachment termed focal adhesions (FAs). In contrast, we demonstrate that DOCK5 recruitment to FAs in Hela cells is restricted by GIT2, an established regulator of FA signaling. We determine that GIT2 is targeted to FAs in response to Rho-ROCK signaling and actomyosin contractility. Accordingly, inhibition of ROCK activity or MLC function promotes enrichment of DOCK5 in membrane protrusions and nascent cell-substratum adhesions. We further demonstrate that GIT2 inhibits the interaction of DOCK5 with Crk. Moreover, we show that depletion of GIT2 promotes DOCK5-dependent activation of the Crk-p130Cas signaling cascade to promote Rac1-mediated lamellipodial protrusion and FA turnover. The antagonism between GIT2 and DOCK5 extends to non-transformed MCF10A mammary epithelial cells, with DOCK5 'dialing-up' and GIT2 'dialing-down' invasiveness. Finally, we determine that DOCK5 inhibition attenuates invasion and metastasis of MDA-MB-231 cells and prolongs life span of mice injected with these cells. Collectively, our work identifies DOCK5 as a key regulator of epithelial invasion and metastasis, and demonstrates that suppression of DOCK5 by GIT2 represents a previously unappreciated mechanism for coordination of Rho and Rac GTPases.

The urokinase receptor associated protein (uPARAP/endo180): a novel internalization receptor connected to the plasminogen activation system.

The urokinase-mediated plasminogen activation system plays a central role in the extracellular proteolytic degradation reactions in cancer invasion. In this review article we discuss a number of recent findings identifying a new cellular receptor protein, uPARAP, that interacts with components of this proteolytic system. uPARAP is a high molecular weight type-1 membrane protein, belonging to the macrophage mannose receptor protein family. On the surface of certain cells, uPARAP forms a ternary complex with the pro-form of the urokinase-type plasminogen activator (uPA) and its primary receptor (uPAR). While the biological consequences of this reaction have not yet been verified experimentally, a likely event is ligand internalization because uPARAP is a constitutively recycling internalization receptor. uPARAP also binds at least one component, collagen type V, in the extracellular matrix meshwork, pointing to a potential role in proteolytic substrate presentation. Additional ligands have been proposed, including collagenase-3 and glycoproteins capable of interacting with one of the multiple carbohydrate recognition-type domains of uPARAP. In various adult tissues uPARAP is present on fibroblasts, macrophages and a subset of endothelial cells. In fetal tissues the protein has also been demonstrated in certain bone forming regions. Hypotheses on the physiological function of uPARAP include regulatory roles in extracellular proteolysis. This type of function would be likely to direct the local turnover of proteases and their substrate degradation products and thus may add to the complicated interplay between several cell types in governing restricted tissue degradation.

Inflammation and uPAR-Expression in Colorectal Liver Metastases in Relation to Growth Pattern and Neo-adjuvant Therapy.

Proteolytic activity and inflammation in the tumour microenvironment affects cancer progression. In colorectal cancer (CRC) liver metastases it has been observed that three different immune profiles are present, as well as proteolytic activity, determined by the expression of urokinase-type plasminogen activator (uPAR).The main objectives of this study were to investigate uPAR expression and the density of macrophages (CD68) and T cells (CD3) as markers of inflammation in resected CRC liver metastases, where patients were neo-adjuvantly treated with chemotherapy with or without the angiogenesis inhibitor bevacizumab. Chemonaive patients served as a control group. The markers were correlated to growth patterns (GP) of liver metastases, i.e. desmoplastic, pushing and replacement GP. It was hypothesised that differences in proteolysis and inflammation could reflect tumour specific growth and therapy related changes in the tumour microenvironment. In chemonaive patients, a significantly higher level of uPAR was observed in desmoplastic liver metastases in comparison to pushing GP (p = 0.01) or replacement GP (p = 0.03). A significantly higher density of CD68 was observed in liver metastases with replacement GP in comparison to those with pushing GP (p = 0.01). In liver metastases from chemo treated patients, CD68 density was significantly higher in desmoplastic GP in comparison to pushing GP (p = 0.03). In chemo and bevacizumab treated patients only a significant lower CD3 expression was observed in liver metastases with a mixed GP than in those with desmoplastic (p = 0.01) or pushing GP (p = 0.05). Expression of uPAR and the density of macrophages at the tumour margin of liver metastasis differ between GP in the untreated patients. A higher density of T cells was observed in the bevacizumab treated patients, when desmoplastic and pushing metastases were compared to liver metastases with a mix of the GP respectively, however no specific correlations between the immune markers of macrophages and T cells or GP of liver metastases could be demonstrated.

Identification of two estrogen regulated genes associated with growth regulation of human breast cancer.

We have identified two estrogen regulated gene products in the E(2) growth inhibited human breast cancer xenograft, T61; one showing 100% homology to the human BAC clone RP11-112E16, the other 100% homology to the human CPR3/DNJ3 gene. Verification by Northern blot analyses showed an up-regulation of the BAC clone RP11-112E16 and the CPR3/DNJ3 mRNAs upon E(2) treatment. Treatment of T61 tumors with tamoxifen, leading to static tumor growth, also increased the expression of the BAC clone RP11-112E16 and the CPR3/DNJ3 mRNAs. A similar association between growth inhibition and BAC clone RP11-112E16 and CPR3/DNJ3 mRNA induction was observed in MCF-7 cells treated with ICI 182.780. In MCF-7 cells, treatment with E(2) resulted in growth stimulation concomitant with a decrease in the BAC clone RP11-112E16 and CPR3/DNJ3 mRNA expression. Treatment with a combination of E(2) and ICI 182.780 abolished the anti-estrogen induced increase in BAC clone RP11-112E16 and CPR3/DNJ3 mRNA expression, indicating that regulation of the gene products is mediated through the ER. The association between growth inhibition and BAC clone RP11-112E16 or CPR3/DNJ3 mRNA expression was supported by high expression of both gene products in brain tissue. Further investigations are ongoing to clarify the biological function of these two gene products.

Differential gene expression in human mammary carcinoma cells: identification of a new member of a receptor family.

Differential gene expression in mammary carcinoma cell lines MCF-7 and MCF-7ADR was investigated by Differential Display Technique. These two cell lines represent a model system for progression of mammary carcinoma: MCF-7 is estradiol-dependent for growth, estrogen-receptor positive, tamoxifen responsive, vimentin negative, Adriamycin sensitive and not invasive in vitro or in vivo while MCF-7ADR is estradiol-independent for growth, estrogen-receptor negative, tamoxifen resistant, vimentin positive, Adriamycin resistant and invasive in vitro and in vivo. Here we describe the identification of a new receptor expressed exclusively in MCF-7ADR. The receptor covers 157 amino acids with a predicted topology of four transmembrane and two extracellular domains. Additional members of the gene family are: peripheral membrane protein PMP22, epithelial membrane protein EMP-1 and squamous cell differentiation marker CL-20.

[Appendicular mucoid impaction disclosing mucoviscidosis with cirrhosis in a young adult without pulmonary lesion]. / Impaction mucoïde appendiculaire révélatrice d'une mucoviscidose accompagnée d'une cirrhose chez un adulte jeune sans lésion pulmonaire.

A 22-year-old man presented with pain in the right iliac fossa. Clinical examination suggested appendicitis and showed splenomegaly. Echography and abdominal CT-sca suggested the diagnosis of cystic fibrosis based on the association of signs of cirrhosis and pancreatic atrophy. The sweat test was positive. The hypothesis of a mucoid appendicular impaction with spontaneous regression was retained based on clinical and radiological signs. This atypical presentation of cystic fibrosis underscores the frequency of obstructive intestinal which occasionally reveals the disease in the adult, and on the absence in this case of otherwise frequently associated problems such as significant pulmonary disease and malnutrition.

[Interest of bilio-pancreatic morphologic study in management of acute pancreatitis (author's transl)]. / Traitement de 30 cas de pancréatite aiguë nécrotico-hémorragique par une dialyse péritonéale médicale.

The interest of morphologic investigations in the diagnosis, the prognosis and the treatment of acute pancreatitis was analyzed in a series of 120 patients. The systematic endoscopy of the biliary tract yields a distinction between the biliary and the non-biliary sources of pancreatitis and can be followed immediately by an endoscopic sphincterotomy. Th morphological study of the bilio-pancreatic area can complete the prognostic indexes defined by the clinical and biological data, document the extension of the intra- and extrapancreatic necrosis and precise the indication and the choice of a surgical treatment.

[Proximal duodenopancreatectomy in the treatment of tumors of the bilio-pancreatic junction]. / La duodénopancréatectomie céphalique dans le traitement des tumeurs du carrefour biliopancréatique.

Between 1978 and 1983, 32 patients underwent a proximal duodeno-pancreatectomy for malignant lesions of the ampullary region on head on the pancreas. Pre-operative diagnosis was made by endoscopic cholangio-pancreatography, echography, tomodensitometry and arteriography. 14 patients underwent a pre-operative endoscopic bile drainage procedure. The operative mortality was 6,2% and the morbidity 43%; the intensity of the icterus as well as the preoperative bile drainage procedure did not change these factors significantly. The pre-operative diagnostic procedures permitted a clear definition of the extent of the tumor and retrospective analysis demonstrated their prognostic significance with regard to long-term survival. 11 patients are still alive. The presence of histologically proven metastases reduce significantly the duration of survival. Surgical resection is the best treatment for malignant tumors of the ampullary region and the head of the pancreas without local or distant metastases.

Mucinous cystadenocarcinoma of the appendix. A rare tumour of the right iliac fossa.

A case of mucinous cystadenocarcinoma of the appendix is presented. The clinical feature is a painful syndrome of the right iliac fossa. In our observation, the diagnosis was not allowed by preoperative imaging. Appendectomy was initially performed and completed by right hemicolectomy and lymphadenectomy after histological diagnosis of the appendicular malignant tumour was forwarded. The prognosis of this tumour is generally excellent providing early diagnosis and wide enough surgery.

[Kidney cancer: fortuitous discovery of a para-uterine mass at the beginning of pregnancy]. / Cancer du rein: découverte fortuite en début de grossesse d'une masse para-utérine.

The authors relate the case of an incidental detection of a renal cell carcinoma in pregnancy. The tumor appeared as an heterogeneous adnexial mass. Special consideration for the diagnostic evaluation was taken because of pregnancy and a minimal number of CAT scan was performed. A radical nephrectomy was performed at 13 weeks. The patient tolerance, the pregnancy and postdelivery courses were as well as possible. No recurrence of the carcinoma occurred. This cas is discussed in the light of the literature.

Quantification of tissue inhibitor of metalloproteinases 2 in plasma from healthy donors and cancer patients.

Tissue inhibitor of metalloproteinases (TIMP)-2 is a highly conserved molecule, which binds both active and latent matrix metalloproteinase (MMP)-2. TIMP-2 is also involved in the activation of MMP-2 on the cell surface. A quantitative enzyme-linked immunosorbent assay (ELISA) was established and optimized for measurement of TIMP-2 in plasma. The capturing antibody in the ELISA was a monoclonal, while the detecting antibody was a chicken polyclonal antibody recognizing the native form of human TIMP-2. The levels of TIMP-2 were measured in ethylenediaminetetraacetic acid (EDTA) and citrate plasma from healthy donors. The median values were determined as 163 ng/ml (n = 186) with a range of 109-253 ng/ml for EDTA plasma and 139 ng/ml (n = 77) with a range of 95-223 ng/ml for citrate plasma. The TIMP-2 concentration in citrate plasma from 15 patients with advanced, stage IV breast cancer had a median value of 160 ng/ml, only slightly higher but statistically distinguishable from the level found in citrate plasma from the healthy donors. In addition, the TIMP-2 concentration in EDTA plasma from colorectal cancer patients revealed a significantly higher level in plasma from patients with Dukes stage A (P = 0.01) compared with patients with more advanced Dukes stages.