Increased adhesion of the promyelocytic leukaemia cell line, NB4, to fibronectin and thrombospondin upon all‐trans‐retinoic acid treatment

Abstract

Summary.We have evaluated the effects of all-trans-retinoic acid (RA) on the adhesion of the human promyelocytic cell line NB4 to various components of the extracellular matrix. NB4 cells, radiolabelled with 111 Indium, showed a 2-3-fold increase (P < 0·001) in adhesion to fibronectin and thrombospondin upon RA (3 × 10 ¹7 M) treatment, whereas adhesion to collagen I, laminin and vitronectin was not modified. The increase in cell adhesion, observed as early as day 1, preceded cell differentiation and was concomitant with tyrosine phosphorylation events. Using flow cytometry, we analysed the expression of major receptors for fibronectin (a4b1 and a5b1) and for thrombospondin (avb3, aIIbb3, CD36 and CD47) on NB4 cells before and after RA treatment. Except for aIIbb3, which was induced on RA-treated cells, we found no significant increase in the expression of the other receptors, and a decrease in the expression of CD36, upon RA treatment. Preincubation of RA-treated cells with blocking antibodies demonstrated a role for a4b1 and a5b1 in cell adhesion to fibronectin and a5b1, aIIbb3, CD36 and CD47 in cell adhesion to thrombospondin. Experiments with the synthetic peptides GRGDS (0·2 mM) and CSVTCG (0·2 mM) confirmed the participation of integrins, and integrins and CD36, in adhesion of RA-treated cells to fibronectin and thrombospondin, respectively. Further inhibition by heparin (10 mg/ml) and/or recombinant heparin-binding domain of thrombospondin (TSP18) indicated the additional participation of heparin-like receptors in cell adhesion to thrombospondin. Our results indicate that increase in NB4 cell adhesion to fibronectin and thrombospondin upon RA treatment is likely to occur through a modulation of the functional state of several receptors for these proteins.

Beneficial effects of retinoic acid on extracellular matrix degradation and attachment behaviour in follicular thyroid carcinoma cell lines

“…First, RA treatment may affect the expression of attachment molecules, such as integrins or E-cadherin. In the literature no consistent effects of RA on integrins are described, some studies reporting increased, unaltered or decreased expression of integrin subclasses v , 3 , 4 , 5 , 1 , 3 , 4 and 5 (Dedhar et al 1991, Morini et al 1999, Pijuan-Thompson et al 1999, Touhami et al 1999. We found no effects of RA on integrin mRNA expression.…”

“…Furthermore, some recent studies report enhanced attachment of tumour cells to extracellular matrix by retinoids (Matarrese et al 1998, Nakagawa et al 1998, Pereda et al 1999, Touhami et al 1999. Alterations in the expression of adhesion molecules have been suggested as an explanation (Matarrese et al 1998, Nakagawa et al 1998, Touhami et al 1999. In thyroid carcinoma cell lines and tissue samples, expression of retinoid receptors has been investigated, suggesting decreased expression of RXR in anaplastic thyroid carcinoma as compared with differentiated thyroid carcinoma cell lines and decreased RXR expression in thyroid carcinoma tissues as compared with normal thyroid tissue .…”

“…One of the explanations may be decreased activity of uPA or tPA or enhanced expression of TIMPs (Hendrix et al 1990. Furthermore, some recent studies report enhanced attachment of tumour cells to extracellular matrix by retinoids (Matarrese et al 1998, Nakagawa et al 1998, Pereda et al 1999, Touhami et al 1999. Alterations in the expression of adhesion molecules have been suggested as an explanation (Matarrese et al 1998, Nakagawa et al 1998, Touhami et al 1999.…”

Low-speed centrifugation of retroviral vectors absorbed to a particulate substrate: a highly effective means of enhancing retroviral titre

“…Pansorbin/fibronectin/retrovirus complexes may also promote infection in a way analogous to RetroNectin, in that cell binding domains in fibronectin may crosslink to the target cells; 16 this may be less effective in NB4 cells since much of the VLA-4 and VLA-5 they express may be inactive. 48 We have used four different batches of Pansorbin, two of Sansorbin and one batch of Pansorbin equivalent from Sigma Aldrich (Poole, UK). While we observed batch variability of two-to four-fold in the concentrating ability of Pansorbin (2000-to 7500-fold increase in titre after concentration), we find it is always more efficient than Sansorbin, which is in turn always better than the Sigma Aldrich product.…”

Orchestration of multiple arrays of signal cross-talk and combinatorial interactions for maturation and cell death: another vision of t(15;17) preleukemic blast and APL-cell maturation

“…ATRA, G-CSF, IFN and PKA-or PKCtargeted extracellular signals, by enhancing both cytokine production and secretion of competent extracellular matrices and expression of adhesion molecules, directly or indirectly induce APL cell responses by hemopoietic cell environment (MartinThouvenin et al, 1992;Marchetti et al, 1996;Peschel et al, 1996;Hsu et al, 1999). ATRA also facilitates NB4 cell adhesion and motility on extracellular matrix (Brown et al, 1999;Touhami et al, 1999;Zang et al, 2000, and our unpublished data) and could ease marrow egress. In conjunction with the secretion of growth-stimulatory cytokines, it could contribute to hyperleukocytosis.…”