Moon, Jin Wook;Chang, Yoon Soo;Han, Chang Hoon;Kang, Shin Myung;Park, Moo Suk;Byun, Min Kwang;Chung, Wou Young;Park, Jae Jun;Yoo, Kyeong Nam;Shin, Ju Hye;Kim, Young Sam;Chang, Joon;Kim, Sung Kyu;Kim, Hee Jung;Kim, Se Kyu
Tuberculosis and Respiratory Diseases
/
v.58
no.4
/
pp.359-366
/
2005
Background : IGFBP-3 inhibits the mitogenic and anti-apoptotic activity of IGF by blocking the binding of IGF to its receptor. However, under certain circumstances, IGFBP-3 can enhance the activity of IGF by protecting IGF from its degradation. More than half of the interindividual variations in IGFBP-3 levels are known to be genetically determined by the polymorphism at -202 locus of IGFBP-3 gene. Method : We attempted to ascertain whether A-202C polymorphic variation of IGFBP-3 gene constitutes a risk factor for non-small cell lung cancer (NSCLC), using PCR-restriction fragment length polymorphism (RFLP). Our study included 104 NSCLC patients and 104 age-, gender-, and smoking status-matched control subjects. Result : In the 104 NSCLC subjects, the genotypic frequencies at the -202 site were as follows: AA = 67 (64.4%), AC = 35 (33.7%), and CC = 2 (1.9%). We did detect significant differences in the genotypic distribution between the NSCLC and the control subjects (p<0.05), and the NSCLC risk correlated significantly with AA genotype at the -202 locus (AA>AC>CC). Using CC genotype as a reference, the odds ratio (OR) for the subjects with AC genotype was 2.60 (95% CI: 0.89 - 8.60), and the OR associated with AA genotype was 5.89 (95% CI: 1.92 - 21.16). Conclusion : These results indicate that the dysregulation of IGF axis should now be considered as another important risk factor for NSCLC, and a potential target for novel antineoplastic therapies and/or preventative strategies in high-risk groups.
The zebrafish is an emerging vertebrate model organism in reproductive biology. The oocyte maturation of zebrafish is triggered by maturation inducing hormone (MIH, 17α,20β-Dihydroxy-4-pregnen-3-one). In almost all animals, the oocyte maturation is governed by activation of pre-MPF which consists of cyclinB and inactive Cdk1. In the oocyte of Xenopus and mice, the activity of Cdk1 is regulated in two ways, one is the interaction with cyclinB and the other is phosphorylation/dephosphorylation of T14/Y15 residues on the Cdk1 by Wee1 and Cdc25. Unlike Xenopus and mice that have a sufficient amount of pre-MPF, pre-MPF is absent in GV oocyte of most teleost including zebrafish. Therefore, the activation of MPF during zebrafish oocyte maturation might totally depend on de novo synthesis of cyclinB proteins. It is reported that the translation of maternal mRNA is regulated by combination of several RNA binding proteins such as CPEB, Dazl, Pum1/Pum2, and insulin-like growth factor2 mRNA-binding protein 3 in the zebrafish oocytes. However, the definitive mechanism of these proteins to regulate the translation of stored maternal mRNAs remains to be elucidated. Therefore, the investigation of the maturation process of the zebrafish oocyte will provide new information that can help identify the role of translational control in early vertebrate oocyte maturation.
This study was conducted to investigate the effects of streptozotocin-induced (STZ) diabetes on insulin-like growth factor-I (IGF-I), insulin-like growth factor binding proteins (IGFBPs), and IGF-I carrier proteins in serum, liver, and kidney. The levels of total and free IGF-I were measured by radioimmunoassay (RIA). The patterns of IGFBPs were determined by western ligand blotting (WLB) analysis. The profiles of IGF-I carrier proteins in serum were determined by column chromatography. The levels of total and free IGF-I in serum were lower in STZ-induced diabetic rat than normal rat (p<0.01). Similarly, the levels of total IGF-I in liver was lowered in STZ-induced diabetic rats. On the other hand, the levels of total IGF-I in kidney were increased in STZ-induced diabetic rats compared with normal rats (p<0.01). In serum and liver from STZ-induced diabetic rats, the amount of IGFBP-3 was decreased and the amount of IGFBP-2 was increased compared with normal rats. There was a not difference in amount of IGFBP-4 in serum between STZ-induced diabetic rats and normal rats. The serums of normal rats have higher 150kDa carrier proteins than in STZ-induced diabetic rats, whereas, most of 50kDa carrier proteins were found in STZ-induced diabetic rats. These results demonstrate that in STZ-induced diabetic rats, IGF-I/IGFBPs system that included functional bioactivity was changed in serum as well as tissues, and these changes may play an important role in pathogenesis of diabetes.
E2 glycoprotein of hepatitis C virus (HCV) comprises a surface of viral particle together with E1 glycoprotein, and is thought to be involved in the attachment of HCV viral particle to receptor (s) on the permissible cells including hepatocytes, B cells, T cells, and monocytes. We constructed a phage library expressing cellular proteins of hepatocytes on the phage surface, which turned out to be 8.8${\times}$$10^5$ cfu of diversity and carried inserts in 95% of library. We screened both cDNA phage library and 12-mer peptide library to identify the cellular proteins binding to E2 protein. Some intracellular proteins including tensin and membrane band 4.1 which are involved in signal transduction of survival and cytoskeleton organization, were selected from cDNA phage library through several rounds of panning and screening. On the contrary, membrane proteins such as CCR7, CKR-L2, and insulin-like growth factor-1 receptor were identified through screening of peptide library. Phages expressing peptides corresponding to those membrane proteins were bound to E2 protein specifically as determined by neutralization of binding assay. Since it is well known that HCV can infect T cells as well as hepatocytes, we examined to see if E2 protein can bind to CCR7, a member of C-protein coupled receptor family expressed on T cells, using CCR7 transfected tells. Human CCR7 cDNA was cloned into pcDNA3.1(-) vector and transfected into human embryonic kidney cell, 293T, and expressed on the surface of the cell as shown by flow cytometer. Binding assay of E2 protein using CCR7 transfected cells indicated that E2 protein bound to CCR7 by dose-dependent mode, giving rise to the possibility that CCR7 might be a putative cellular receptor for HCV.
Kim, Sun Young;Kim, Se Rim;Lee, Jung Chang;Yi, Ho Keun;Lee, Dae Yeol;Hwang, Pyoung Han
Clinical and Experimental Pediatrics
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v.49
no.4
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pp.431-438
/
2006
Purpose : Insulin-like growth factor binding protein(IGFBP)-3 has been known as a tumor suppressor gene, and its anti-tumor function was divided into insulin-like growth factor(IGF)-dependent and IGF-independent mechanism. In IGF-independent mechanism, IGFBP-3 directly interacts with a cell without binding of IGFs, becoming an interesting object in oncology. Several studies demonstrate that one of the well-known tumor suppressor genes, p53, induces directly IGFBP-3 transcription, and the increment of IGFBP-3 expression induces apoptosis of many cancer cells. Recently, the anti-tumor mechanisms of IGFBP-3 have been reported, but post-translational modification of IGFBP-3 and its anti-tumor mechanism are not well known. In this study, we examined whether p53 regulated the glycosylation of IGFBP-3, and analysed the meaning of IGFBP-3 glycosylation related to the apoptosis of cancer cell. Methods : The p53-mutated status of MDA-MB-231 human breast cancer cells was used in this experiment. The expression and glycosylation of IGFBP-3 were tested by Western blot analysis after infection of adenovirus mediated Ad/p53 and/or Ad/IGFBP-3. Results : Ad/p53 infected cells resulted in growth retardation and the induced apoptosis. p53 induced direct expression and glycosylation of IGFBP-3. The increase of glcosylated IGFBP-3 was able to promote cellular apoptosis, and the glycosylation of IGFBP-3 was more activated by the double treatment of Ad/p53 and Ad/IGFBP-3. Conclusion : From this study, the anti-tumor activity of IGFBP-3 was shown to improve the stabilization of IGFBP-3 through the increment of glycosylation of IGFBP-3 by p53. This result suggests that the combined gene therapy of p53 and IGFBP-3 may appropriate treatment of cancer.
Yun, J.S.;Kang, W.J.;Seo, D.S.;Lee, C.Y.;Oh, S.;Ko, Y.
Asian-Australasian Journal of Animal Sciences
/
v.16
no.4
/
pp.481-488
/
2003
Insulin-like Growth Factors (IGFs) and IGF-binding protein act as intra-ovarian regulators that modulate the proliferation and differentiation of the granulosa and theca cells. Moreover, the IGF system is involved in metabolism by modulating the synthesis and degradation of glycogen and protein in animals. However the effect of the IGF system on egg productivity or body growth in KNOC has not been studied in depth. Therefore, this study was performed to investigate differences of serum IGFs and binding protein expressions between two groups showing high and low egg production or body weight and to elucidate the relationship of IGFs with egg productivity and body growth. KNOCs were divided into high and low groups depending on their egg productivity or body growth, and sera were collected every 10 wk from 20 till 60 wk. Serum IGF-I and -II concentration were measured by RIA using human and mouse antiserum and chicken standards. IGFBP was detected by Western ligand blotting. IGF-I concentrations were significantly greater in the high egg production group compared with those in the low egg production group (30 wk, p<0.01; 20 and 40 wk, p<0.05). Also, differences in IGF-II amounts between the two groups were detected at 60 wk (p<0.05). But IGFBPs in the low egg production group were more intense than that in the high egg production group through the egg laying period. The correlation between IGF-I concentration and number of egg production is significantly positive (20 wk, r=0.2729: p<0.05; 40 wk, r=0.3500: p<0.01), while IGF-II shows no correlation with egg productivity. In male KNOC, IGF-I and -II concentrations in the high body weight group are lower than that in the low body weight group. Body weight also shows a negative correlation with the serum IGF-II concentration in male chickens (20 wk, r=-0.5901: p<0.01). Consequently, we suggest that IGFs and binding protein are (in)directly involved in the egg productivity and body growth in KNOC.
The insulin-like growth factor (IGF)s are believed to one of several growth factors that play an adjunctive role in ovarian follicular development. These factors circulate bound to a family of IGF-binding protein (IGFBP)s. It is known that circulating IGFBPs are involved in the transport of IGFs to tissues and modulate IGFs actions at local tissue. The purposes of this study were to evaluate changes in serum IGFBPs profiles during normal ovulatory menstrual cylce and to compare serum IGFBPs profiles in periovulatory phase of between normal ovulatory menstrual cylce and controlled hyperstimulated cycle. Fasting blood samples were obtained from 15 normal healthy women throughout normal ovulatory menstural cyle and on the day of aspiration of oocyte from 10 patients undergoing ovarian hyperstimuation for in vito fertilization-embryo transfer. Serum IGFBP-1 - IGFBP-4 were measured by western ligand blot and immunoprecipitation. Serum $17{\beta}$-estradiol was determined by radioimmunoassay. Type and molecular weight of serum IGFBP did not changed during normal ovulatory menstural cycle. No significant variation in the relative proportion and level of each IGFBP was found throughout normal ovulatory menstural cyle. Also, the relative proportion and level of each IGFBP did not correlated with serum $17{\beta}$-estradiol level. There was no significant difference in the relative proportion and level of each serum IGFBP between on the day of ovulation in normal ovulatory menstrual cylce and on the day of aspiration of oocyte in controlled hyperstimulated cycle. Our data indicate that IGFBPs have regulatory functions in ovary through an paracrine and autocrine rather than endocrine mechanism during normal ovulatory menstural cycle.
This study examined the effect of insulin-like growth factor (IGF)-I expression in the liver and muscle on the growth of Paralichthys olivaceus fed diets low in fish meal. A feeding experiment was conducted at Jeju National University, Jeju Island, Korea. Groups of P. olivaceus (total initial weight: 200 g) were maintained for 20 weeks on one of five experimental diets containing different proportions of fish meal. Diets containing 0%, 20%, 30%, 40%, and 50% fish meal were labeled FM0, FM20, FM30, FM40, and FM50, respectively. Fish growth was observed every 4 weeks during the feeding experiment, and plasma and liver and muscle tissues were sampled. Plasma IGF-I levels were analyzed using an ELISA kit. The mechanism of IGF-I receptor signaling was examined using immunoblotting and reverse transcription-polymerase chain reaction. The greatest total weight increase was observed in the FM30 group. In parallel, plasma levels of IGF-I and IGF-binding protein were highest in the FM30 group, and mRNA and protein expression were also significantly higher in this group. The first step in the IGF-I signaling pathway, tyrosine-phosphorylation checking, occurred smoothly until 20 weeks. These results suggest that a dietary ratio of 30% fish meal best promotes growth in this species. The IGF-I signaling pathway in the liver and muscle is associated with growth in P. olivaceus.
Purpose: This pilot study assessed changes in the growth plate and growth rates in children during a 6-month period. Methods: The study included 31 healthy children (17 boys, 14 girls) under evaluation for growth retardation. Height, weight, bone age, insulin like growth factor-1 (IGF-1), and insulin like growth factor binding protein 3 (IGF-BP3) were measured at baseline and after 6 months. In addition, the diameter, thickness, and volume of the femoral and tibial growth plates were measured using magnetic resonance imaging. Results: The mean bone age in boys and girls was 11.7 and 10.7 years, respectively. In boys, height (z score) (-0.2 vs. 0.0), weight (z score) (0.8 vs. 1.1), body mass index (BMI) (z score) (1.27 vs. 1.5), IGF-1 (ng/mL) (343.6 vs. 501.8), and IGF-BP3 (ng/mL) (5,088.5 vs. 5,620.0) were significantly higher after 6 months. In girls, height (z score) (-1.0 vs. -0.7), weight (z score) (-0.5 vs. 0.1), BMI (z score) (-0.02 vs. 0.3), IGF-1 (ng/mL) (329.3 vs. 524.6), and IGF-BP3 (ng/mL) (4,644.4 vs. 5,593.6) were also significantly higher after 6 months. In both sexes, the mean diameter and volume of the femoral and tibial growth plates were significantly increased 6 months later. Conclusion: No significant correlation was found between changes in the growth plate and clinical parameters in children with growth retardation in this study, other than correlations of change in femoral diameter with weight and BMI. A larger, long-term study is needed to precisely evaluate the correlation between change in the growth plate and growth.
The somatotropic axis plays a key role in proliferation and differentiation of avian organs during both pre- and posthatching periods. This review discusses the complexity of regulation of the endocrine system for chicken development and growth by growth hormone (GH), insulin-like growth factor (IGF), and IGF binding protein (IGFBP). In addition, the thyrotropic axis, including thyrotropin-releasing hormone (TRH) and thyroid hormones ($T_4$ and $T_3$), is also involved in the GH-secreting pattern. In mammals, IGFI and -II are always sequestered in a 150 kDa non-covalent ternary complex. This complex consists of one molecule each of IGF-I or IGF-II, IGFBP-3 or IGFBP-5 and an acid labile subunit (ALS). Chick ALS is identified in different strains for the first time, and further investigation of the expression of ALS on developmental stage and ALS effect on IGF bioavailability may be addressed in the future.
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