|Insulin-like growth factor binding proteins 4 and 7 released by senescent cells promote premature senescence in mesenchymal stem cells.|
Severino, V; Alessio, N; Farina, A; Sandomenico, A; Cipollaro, M; Peluso, G; Galderisi, U; Chambery, A
Cell death & disease
Cellular senescence is the permanent arrest of cell cycle, physiologically related to aging and aging-associated diseases. Senescence is also recognized as a mechanism for limiting the regenerative potential of stem cells and to protect cells from cancer development. The senescence program is realized through autocrine/paracrine pathways based on the activation of a peculiar senescence-associated secretory phenotype (SASP). We show here that conditioned media (CM) of senescent mesenchymal stem cells (MSCs) contain a set of secreted factors that are able to induce a full senescence response in young cells. To delineate a hallmark of stem cells SASP, we have characterized the factors secreted by senescent MSC identifying insulin-like growth factor binding proteins 4 and 7 (IGFBP4 and IGFBP7) as key components needed for triggering senescence in young MSC. The pro-senescent effects of IGFBP4 and IGFBP7 are reversed by single or simultaneous immunodepletion of either proteins from senescent-CM. The blocking of IGFBP4/7 also reduces apoptosis and promotes cell growth, suggesting that they may have a pleiotropic effect on MSC biology. Furthermore, the simultaneous addition of rIGFBP4/7 increased senescence and induced apoptosis in young MSC. Collectively, these results suggest the occurrence of novel-secreted factors regulating MSC cellular senescence of potential importance for regenerative medicine and cancer therapy.
|Estradiol increases relative amounts of insulin-like growth factor binding protein (IGFBP)-3 in serum and expression of IGFBP-2 in anterior pituitaries of ewes.|
J A Clapper, J L Snyder, A J Roberts, D L Hamernik, G E Moss
Biology of reproduction
This study determined whether estradiol regulates insulin-like growth factor (IGF)-I and IGF binding proteins (IGFBPs) in the pituitary gland, hypophyseal stalk median eminence (SME), and circulation concomitantly with effects on LH. Ovariectomized ewes received an estradiol implant or no implant during the anestrous season and were slaughtered 80 days later. Estradiol suppressed serum LH to a greater extent during anestrus than after onset of the breeding season (Days 60 and 75). Amounts of mRNA for LHbeta subunit were decreased by estradiol, but mRNA for alpha and FSHbeta subunits were not affected. Estradiol increased serum IGF-I, IGFBP-3, and IGFBP-4 throughout the treatment period, but it did not influence other IGFBPs in serum. In response to estradiol, pituitary IGFBP-2 tended to increase and mRNA for IGFBP-2 increased twofold. Other IGFBPs in the pituitary gland were not influenced by estradiol. In the SME, IGFBP-2, IGFBP-5, and the 40-kDa IGFBP-3 were increased by estradiol. Thus, estradiol influences both the IGF and gonadotropin systems in sheep. Estradiol influences on gonadotroph function may be mediated by alterations in the IGF system.
|Identification of the forms of insulin-like growth factor-binding proteins produced by human fibroblasts and the mechanisms that regulate their secretion.|
Camacho-Hubner, C, et al.
J. Biol. Chem., 267: 11949-56 (1992)
Human fibroblasts secrete insulin-like growth factor-binding proteins (IGFBPs) that can modify insulin-like growth factor (IGF) I action. We have determined the molecular identities of three forms of IGFBPs that are secreted by human fibroblasts in vitro. Ligand blot analysis of fibroblast conditioned media revealed that the M(r) 43,000 and 39,000 forms were the most abundant, but that M(r) 31,000 and 24,000 forms were also present. An antiserum that was specific for IGFBP-5 reacted with the M(r) 31,000 form, and an IGFBP-4-specific antiserum recognized only the M(r) 24,000 form. The M(r) 39,000 and 43,000 forms were detected by IGFBP-3 antiserum. Further proof that fibroblasts synthesized these forms of IGFBPs was obtained by Northern blotting. A cDNA probe for IGFBP-3 hybridized with a 2.4-kilobase (kb) transcript, whereas a cDNA probe for IGFBP-5 recognized a single 6.0-kb transcript, and an IGFBP-4 cDNA probe recognized 2.2- and 2.0-kb transcripts. IGF-I and -II caused a minimal (less than 43%) increase in IGFBP-5 mRNA abundance and had no effect on IGFBP-4 mRNA abundance. IGF-I and -II (100 ng/ml) stimulated 6-8-fold increases in IGFBP-5 levels, whereas IGFBP-4 was inhibited. Insulin failed to elicit any change in IGFBP-5, suggesting that binding of the IGFs to IGFBPs was required to detect the increase. Immunoblotting for IGFBP-5 revealed an M(r) 23,000 (non-IGF-I-binding) fragment. To determine if the IGFs were influencing proteolytic degradation of IGFBP-5, pure IGFBP-5 was added to fibroblast cultures and incubated for 4 h at 37 degrees C. The amount of fragment formation was attenuated by the presence of IGF-I and -II, but not insulin, suggesting that this is a mechanism by which the IGFs act to modulate IGFBP-5 concentration. In contrast to the IGFs, forskolin, which increased IGFBP-4 and -5 mRNA abundance and secretion, had no effect on fragment formation. The results show that human fibroblasts synthesize and secrete IGFBP-3, -4, and -5 and that changes in intracellular cAMP regulate synthesis, whereas the IGFs regulate IGFBP-4 and -5 levels by post-transcriptional mechanisms.