Different adhesion properties of highly and poorly metastatic HT-29 colon carcinoma cells with extracellular matrix components: role of integrin expression and cytoskeletal components

Abstract

Summary Integrin-mediated tumour cell adhesion to extracellular matrix (ECM) components is an important step in the development of metastatic lesions. Thus, integrin expression and integrin-mediated adhesion of colon carcinoma cells to various ECM components was examined. Poorly (HT-29P) and highly (HT-29LMM) liver-metastatic colon carcinoma cells were used to study the rates of adhesion to collagen I (C I), collagen IV (C IV), laminin (LN), fibronectin (FN), or vitronectin (VN) in a static adhesion assay (10-120 min). Cells were untreated or treated with oligopeptides (RGD, GRGDS, YIGSR, RGES), anti-integrin antibodies, or colchicine, nocodazole, cycloheximide, acrylamide or cytochalasin D (to disrupt cytoskeletal structures). Both cell lines expressed similar patterns of integrin expression (α 2 , α 3 , α 6 , α v , β 1 , β 4 and β 5 ) by immunocytochemistry and immunoprecipitation. HT-29LMM cells showed significantly higher rates of adhesion to LN (P < 0.001) and FN (P < 0.001), but significantly poorer rates of adhesion to C I (P < 0.05) and C IV (P < 0.001) than HT-29P cells, respectively, adhesion to VN was insignificant. RGD and GRGDS inhibited HT-29LMM cell adhesion to FN only. Pretreatment with anti-β 1 or anti-α 2 integrin subunits suppressed adhesion to C I and C IV, and adhesion to LN was inhibited with anti-β 1 or anti-α 6 integrin. Anti-β 1 or anti-α v blocked adhesion to FN. Pretreatment of cells with cytochalasin D, cycloheximide or acrylamide inhibited adhesive interactions of both cell lines to the ECM components. In contrast, colchicine and nocodazole had no effect. The results demonstrate that adhesion of HT-29 cells to ECM is mediated, in part, by different integrins, depending on the substrate. Poorly and highly metastatic HT-29 cells possessed different patterns of adhesion to the various ECM substrates, but these differences were not due to different expression of integrin subunits. The results also suggested that the initial adhesion of poorly or highly metastatic HT-29 cells to ECM components requires, in part, the presence of native action and intermediate filaments, but not of microtubules. Thus the adhesion of tumour cells to ECM components may be dependent on signal transduction and assembly of microfilaments.

Rotavirus Replication in Intestinal Cells Differentially Regulates Integrin Expression by a Phosphatidylinositol 3-Kinase-Dependent Pathway, Resulting in Increased Cell Adhesion and Virus Yield

“…This antibody reduced collagen adherence by 29% (Caco-2), 48% (HT-29), and 53% (MA104) for mock-infected cells and 38% (Caco-2), 57% (HT-29), and 39% (MA104) for RRV-infected cells. Adhesion of uninfected Caco-2 and HT-29 cells to type 1 collagen was similarly reduced by anti-␣2 antibodies in previous studies (33,36,50). Our experiments show this function is retained in rotavirusinfected cells.…”

“…The ␣2␤1, ␣V␤3, ␣V␤5, ␣5, and ␤2 integrins were chosen for analysis based on their predominance on rotavirus-permissive intestinal cells (33,36,42,50,74) and the role of a subset in rotavirus cell entry. It is demonstrated that rotavirus infection of intestinal cells differentially regulates rotavirus receptor and non-receptor integrin mRNA and protein expression by a PI3K-dependent pathway that is activated by infection.…”

Mechanisms regulating colorectal cancer cell metastasis into liver (Review)

“…Due to an incomplete layer of hepatic ECs, these ECM components are directly accessible to circulating cells (16,18). Various studies have shown that interactions between these ECM components within the liver and tumor cells appear to be crucial to the formation of hepatic metastases (19)(20)(21). The liver is furthermore unique, since it lacks a basement membrane under the sinusoidal endothelium (22).…”

Functional differences between two morphologically distinct cell subpopulations within a human colorectal carcinoma cell line

“…Several mechanisms are implicated in the process, including changes in adhesion properties that allow tumor cell detachment and migration (9,10). Adhesion of metastatic cells to the underlying extracellular matrix is very important (11), and the acquisition of metastatic competence by clones of cells with growth advantages is thought to be associated with an ability to coordinate the adhesive properties in every step of metastasis.…”