Anti-angiogenic effects of thalidomide: expression of apoptosis-inducible active-caspase-3 in a three-dimensional collagen gel culture of aorta

Abstract

The anti-angiogenic properties of thalidomide have led to the use of the agent as a remedy for multiple myeloma. Nevertheless, the anti-angiogenic moiety of thalidomide remains unidentified. In this study we examined the anti-angiogenic effects of thalidomide in an in vitro model using a three-dimensional collagen gel culture. Angiogenesis was significantly inhibited when the culture was treated with thalidomide plus cytochrome P-450 (CYP2B4), and the migrating cells and tubules were positive for active-caspase-3 in an accompanying immunohistochemical investigation. Transmission electron microscopic observation also confirmed that active-caspase-3-positive cells demonstrated apoptotic characteristics. This study is the first to morphologically demonstrate the effect of thalidomide in directly inducing the apoptosis of new tubules and migrating cells on a three-dimensional collagen gel culture of aorta. Taken together with earlier findings, our new results indicate that the thalidomide-induced inhibition of angiogenesis involves apoptosis in addition to the suppression of TNF-alpha and inhibition of cell migration from aorta explants, i.e., the factors important for capillarogenesis.

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“…Accordingly, the drug has been reintroduced into clinical therapy for the treatment of multiple myeloma and several other types of cancer (for a review, see Fujita et al 2004). The proposed anti-angiogenic effects of thalidomide were investigated by Fujita et al (2004) using a three-dimensional culture system for capillary sprouts from mouse aortae and a human in vitro model with human umbilical vein endothelial cells (HUVECs) and fibroblasts.…”

“…Accordingly, the drug has been reintroduced into clinical therapy for the treatment of multiple myeloma and several other types of cancer (for a review, see Fujita et al 2004). The proposed anti-angiogenic effects of thalidomide were investigated by Fujita et al (2004) using a three-dimensional culture system for capillary sprouts from mouse aortae and a human in vitro model with human umbilical vein endothelial cells (HUVECs) and fibroblasts. In convincing immunohistochemical and electron microscopic images, the authors for the first time demonstrated the effect of thalidomide in directly promoting apoptosis (characterized by the presence of activecaspase-3) of sprouting endothelial tubules and of migrating cells in the aorta cultures, possibly attributable to thalidomide-linked ROS formation.…”

DNA Micro-Array Gene Expression Profiling of Angiogenesis in Collagen Gel Culture

“…The culture of aortic explant in collagen gels has been used widely and effectively for analyzing the biological process of angiogenesis (Mori et al 1988;Nicosia and Ottinetti, 1990;Akita et al 1997a;Akita et al 1997b;Fujita et al 2002;Nagatoro et al 2003;Fujita et al 2004;Suda et al 2004;Fujita et al 2006). Our previous paper showed that the capillary tubes formed in the collagen gel had architecture similar to that of capillaries in vivo observed by electron microscopy (Akita et al 1997a;Akita et al 1997b).…”

“…This collagen culture technique is a modifi cation of our previous work (Nagatoro et al 2003;Fujita et al 2004). The thoracic aorta samples were obtained from 1 month old ICR male mice (n = 16).…”

Mechanism of immunomodulatory drugs' action in the treatment of multiple myeloma

“…Thus, the decreased NF-kB activity or the decreased expression of IL-6 by IMiD will also play a pro-apoptotic role. Furthermore, the fact that IMiDs triggered activation of caspase 3 [88,138,[156][157][158][159][160][161][162][163][164][165][166][167], caspase 8 [119,158,[162][163][164]167,168], caspase 9 [138,158,159,[162][163][164][165]168], and caspase 12 [163], the increased expression of pro-apoptotic factors BAX and BAK [169] and the decreased expression of anti-apoptotic factors Bcl-2 [169], cFLIP, and Bcl-xL [170] indicates that IMiDs possess significant pro-apoptotic effects.…”