In vivo and in vitro invasion in relation to phenotypic characteristics of human colorectal carcinoma cells

Vries, J. E. De; Dinjens, Winand N.M.; Bruyne, Georges K. De; Verspaget, H. W.; Linden, Edith P.M. van der; Bruïne, Adriaan P. de; Mareel, Marcus; Bosman, F.; Kate, Joop ten

doi:10.1038/bjc.1995.55

Cited by 47 publications

(26 citation statements)

References 32 publications

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“…Although Caco-2 cells are commonly used to model diverse aspects of nonmalignant intestinal epithelial biology and signal transduction (36,63), they are nevertheless a transformed cell line originally derived from a colon cancer (12,19). We therefore confirmed our results in nontransformed small intestinal epithelial IEC-6 cells (Fig.…”

Section: Resultssupporting

confidence: 80%

ILK mediates the effects of strain on intestinal epithelial wound closure

Yuan

Sanders

Basson

2011

American Journal of Physiology-Cell Physiology

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The intestinal epithelium is subjected to repetitive deformation during normal gut function by peristalsis and villous motility. Such repetitive strain promotes intestinal epithelial migration across fibronectin in vitro, but signaling mediators for this are poorly understood. We hypothesized that integrin-linked kinase (ILK) mediates strain-stimulated migration in intestinal epithelial cells cultured on fibronectin. ILK kinase activity increased rapidly 5 min after strain induction in both Caco-2 and intestinal epithelial cell-6 (IEC-6) cells. Wound closure in response to strain was reduced in ILK small interfering RNA (siRNA)-transfected Caco-2 cell monolayers when compared with control siRNA-transfected Caco-2 cells. Pharmacological blockade of phosphatidylinositol-3 kinase (PI3K) or Src or reducing Src by siRNA prevented strain activation of ILK. ILK coimmunoprecipitated with focal adhesion kinase (FAK), and this association was decreased by mutation of FAK Tyr925 but not FAK Tyr397. Strain induction of FAK Tyr925 phosphorylation but not FAK Tyr397 or FAK Tyr576 phosphorylation was blocked in ILK siRNA-transfected cells. ILK-Src association was stimulated by strain and was blocked by the Src inhibitor PP2. Finally, ILK reduction by siRNA inhibited strain-induced phosphorylation of myosin light chain and Akt. These results suggest a strain-dependent signaling pathway in which ILK association with FAK and Src mediates the subsequent downstream strain-induced motogenic response and suggest that ILK induction by repetitive deformation may contribute to recovery from mucosal injury and restoration of the mucosal barrier in patients with prolonged ileus. ILK may therefore be an important target for intervention to maintain the mucosa in such patients.

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Section: Resultssupporting

confidence: 80%

ILK mediates the effects of strain on intestinal epithelial wound closure

Yuan

Sanders

Basson

2011

American Journal of Physiology-Cell Physiology

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“…In the subcutis transplanted tumour cells rarely show invasive growth, due to restrictions of the local stromal environment. In contrast, in orthotopic transplantation (in the caecum) the tumour cells are confronted with organ-specific stromal cells (de Vries et al, 1995). Our results show that all colon carcinoma cells that grow as caecal xenografts are also invasive in vivo.…”

Section: Discussionmentioning

confidence: 59%

A comparative evaluation of various invasion assays testing colon carcinoma cell lines

et al. 1999

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Summary Various colon carcinoma cell lines were tested in different invasion assays, i.e. invasion into Matrigel, into confluent fibroblast layers and into chicken heart tissue. Furthermore, invasive capacity and metastatic potential were determined in nude mice. The colon carcinoma cells used were the human cell lines Caco-2, SW-480, SW-620 and HT-29, and the murine lines . None of the human colon carcinoma cells migrated through porous membranes coated with Matrigel; of the murine lines, only Colon-26 did. When incubated in a mixture of Matrigel and culture medium non-invading cells formed spheroid cultures, whereas invading cells showed a stellate outgrowth. Only the heterogeneously shaped (epithelioid and stellate) cells of SW-480 and SW-620 and the spindle-shaped cells of Colon-26 invaded clearly confluent skin and colon fibroblasts as well as chicken heart tissue. However, when transplanted into the caecum of nude and syngeneic mice, all the lines tested were invasive with the exception of Caco-2 cells. We conclude that the outcome of in vitro tests measuring the invasive capacity of neoplastic cells is largely dependent on the test system used. Invasive capacity in vitro is strongly correlated with cells having a spindle cell shape, vimentin expression and E-cadherin down regulation. In contrast, HT-29 and Colon-38 cells having an epithelioid phenotype were clearly invasive and metastatic in vivo, but not in vitro. © 1999 Cancer Research Campaign Keywords: Matrigel; confluent fibroblast layers; chicken hearts; orthotopic transplantation 934British Journal of Cancer (1999) 81(6), 934-941 © 1999 Cancer Research Campaign Article no. bjoc.1999 Received 2 In vitro testing Matrigel assayThe assay was carried out as described by Albini et al (1987). Perforated polyvinylpyrrolidone-free polycarbonate membranes of 13-mm diameter (Nucleopore, CA, USA) were used with 8-µm pore size, which is sufficiently large to allow the passage of single carcinoma cells.To promote attachment all the membranes were first coated on the lower side in 4-well dishes with 15 µl of a 0.1% solution of fibronectin (Sigma, St Louis, MO, USA) in serum-free medium according to Sieuwerts et al (1997). Subsequently half of the membranes were coated with a layer of Matrigel to measure invasion. An amount of 20 µl containing 40 µg Matrigel was applied per membrane surface obtained by dilution of a stock solution with serum-free medium as recommended (Collaborative Biomedical Products, Becton and Dickinson Labware, Bedford, UK). The concentration used prevents preliminary detachment of the Matrigel from the filters and falls within the range of concentrations used by others (see Discussion). The Matrigel was gelatinized at 37°C for 20 min. The other half of the membranes was left uncoated to measure migration.The membranes were sealed in a sterilized Boyden chemotaxis chamber (Nucleopore Membrane Products, CA, USA) after filling the lower compartment with 220 µl of serum-free NIH-3T3 conditioned medium as attractant. The upper compartment was ...

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“…34 We also excluded N-terminal protein truncation due to mutations or alternative mRNA splicing and destabilization of EGF domains through Ca 2ϩ inaccessibility 10 as mechanisms that might impair binding of CD97 EGF mAbs to smooth muscle cells. In addition, CD97 stalk mAbs did not crossreact with the related EGF-TM7 receptor ETL 35 expressed by smooth muscle (data not shown).…”

Section: Discussionmentioning

confidence: 92%

N‐glycosylation of CD97 within the EGF domains is crucial for epitope accessibility in normal and malignant cells as well as CD55 ligand binding

Wobus

Vogel

Schmücking

et al. 2004

Intl Journal of Cancer

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CD97 is an EGF-TM7 receptor found on various carcinomas where expression levels correlate with dedifferentiation and tumor stage, smooth muscle cells and leukocytes. CD97 acts as an adhesion molecule by binding to its cellular ligand, CD55. In this study, we demonstrate that 2 immunodominant CD97 epitopes are not equally present in the various cell types. Differences were apparent in gastrointestinal tumors and smooth muscle cells where monoclonal antibodies (mAbs) to the first epidermal growth factor (EGF) domain (CD97 EGF ) showed a more restricted staining pattern than mAbs to the stalk region (CD97 stalk ). This discrepancy was not detectable in cultured gastrointestinal tumor cell lines. In fact, the selection of the CD97 mAb influences the result of clinical studies. Thus, we clarified the reason(s) for these differences in CD97 mAb staining on various cell types. We provide evidence that epitope accessibility for CD97 EGF Key words: CD97; colorectal cancer; smooth muscle cell; epitope accessibility; glycosylation CD97 is a member of a subfamily of 7-span transmembrane (TM7) receptors referred to as EGF-TM7 proteins. 1,2 In normal tissues, CD97 is abundantly expressed in smooth muscle cells, macrophages, granulocytes, dendritic cells and in some T and B cells. 3 Carcinomas of different origin showed higher CD97 expression compared to the corresponding normal cell types. 4 -7 CD97 consists of an extended extracellular region with several N-terminal epidermal growth factor (EGF) domains coupled to the TM7 domain by a stalk. 1,8 Various CD97 isoforms exist possessing 3 (EGF 1,2,5), 4 (EGF 1,2,3,5), or 5 (EGF 1,2,3,4,5) EGF domains. CD97 binds to membrane receptor CD55. 9 This interaction is mediated by the first 2 CD97 EGF domains. 9 -11 Chondroitin sulfate glycosaminoglycan, present in cell membranes and the extracellular matrix, is a new ligand to the longest CD97 isoform (EGF 1,2,3,4,5). 12 The ability of CD97 EGF domains to interact with cellular and/or extracellular matrix ligands suggests that the molecule is involved in cell-cell or cell-matrix interactions. Various lines of evidence have shown an important role for CD97 in tumor dedifferentiation, migration, invasiveness and metastasis. CD97 levels correlate with the in vitro migration and invasion capacity of colorectal tumor cell lines and CD97-transfected cells. 6 Colorectal carcinomas with more strongly CD97-stained, scattered tumor cells at the invasion front showed a poorer clinical stage as well as increased lymph vessel invasion compared to cases with uniform CD97 staining. 6 In thyroid cancers, CD97 expression parallels the aggressiveness and lymph node involvement of these tumors. 4,5 Our previous data revealed that CD97 has prognostic and therapeutic potential in cancer. However, analyzing CD97 expression brings up the phenomenon that different staining patterns, depending on which CD97 monoclonal antibody (mAb) has been used for immunohistology, can be readily observed in pancreatic tissues. 7,13 Based on this observation, one aim of ...

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In vivo and in vitro invasion in relation to phenotypic characteristics of human colorectal carcinoma cells

Cited by 47 publications

References 32 publications

ILK mediates the effects of strain on intestinal epithelial wound closure

ILK mediates the effects of strain on intestinal epithelial wound closure

A comparative evaluation of various invasion assays testing colon carcinoma cell lines

N‐glycosylation of CD97 within the EGF domains is crucial for epitope accessibility in normal and malignant cells as well as CD55 ligand binding

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