Interaction of endothelial cells with a laminin A chain peptide (SIKVAV) in vitro and induction of angiogenic behavior in vivo

Abstract

Endothelial cells are known to bind to laminin, and two peptides derived from the laminin A (CTFALRGDNP) and B1 (CDPGYIGSR) chains block the capillary-like tube formation on a laminin-rich basement membrane matrix, Matrigel. In the present study, we have used various in vitro and in vivo assays to investigate the angiogenic-biologic effects of a third active site in the laminin A chain, CSRARKQAASIKVAVSADR (designated PA22-2) on endothelial cells. The SIKVAV-containing peptide was as active as the YIGSR-containing peptide for endothelial cell attachment but was less active than either the RGD-containing peptide or intact laminin. Endothelial cells seeded on this peptide appeared fibroblastic with many extended processes, unlike the normal cobblestone morphology observed on tissue culture plastic. In addition, in contrast to normal tube formation on Matrigel, short irregular structures formed, some of which penetrated the matrix and sprouting was more apparent. Analysis of endothelial cell conditioned media of cells cultured in the presence of this peptide indicated degradation of the Matrigel and zymograms demonstrated active collagenase IV (gelatinase) at 68 and 62 Kd. A murine in vivo angiogenesis assay and the chick yolk sac/chorioallantoic membrane assays with the peptide demonstrated increased endothelial cell mobilization, capillary branching, and vessel formation. These data suggest that the -SIKVAV-site may play an important role in initiating branching and formation of new capillaries from the parent vessels, a behavior that is observed in vivo in response to tumor growth or in the normal vascular response to injury.

PI4KIIα is a novel regulator of tumor growth by its action on angiogenesis and HIF-1α regulation

“…Three measurements taken randomly were obtained for each treatment and the experiment was repeated three times. The endothelial tube formation assay was performed as described previously with the following modifications (Grant et al, 1992). HUVECs were cultured in either of the HeLa cell media described above.…”

6‐(1‐Oxobutyl)‐5,8‐dimethoxy‐1,4‐naphthoquinone inhibits lewis lung cancer by antiangiogenesis and apoptosis

“…An in vitro capillary differentiation assay was performed on Matrigel as originally described by Grant et al 25 We have used this method in previous works. [19][20][21] In experiments where LLCconditioned culture media were used as the source of angiogenic stimulation in the capillary tube formation assay, LLC cells were washed with phosphate-buffered saline (PBS) and then cultured in fresh RPMI 1640 medium with or without OXO, under either normoxic or hypoxic condition for 12 hr, and then the media were collected.…”

Hydrogel scaffolds composed of genetically synthesized self-assembling peptides for three-dimensional cell culture

“…EREI, which consists of ERE fused with the IKVAV peptide, not only exhibits cell adhesive activity but also promotes angiogenesis and neural differentiation as a result of the combination of IKVAV and RGD. [8][9][10][11][12] In our genetic engineering-based designs, multi-functional ECMs composed not only of peptide motifs but also of growth factors were constructed. Epidermal growth factor fused to the ERE protein was found to exhibit cell growth activity via the epidermal growth factor moiety.…”