노인에서 IGFBP-3 Promoter Polymorphism이 제2형 당뇨병에 미치는 영향

박종숙(Jong-Suk Park),남주영(Joo-Young Nam),김철식(Chul-Sik Kim),김똘미(Dol-Mi Kim),조민호(Min-Ho Cho),박진아(Jina Park),안철우(Chul-Woo Ahn),차봉수(Bong-Soo Cha),임승길(Sung-Kil Lim),김경래(Kyung-Rae Kim),이현철(Hyun-Chul Lee) 대한임상노인의학회 2004 대한임상노인의학회지 Vol.5 No.2

연구배경: Insulin-like growth factor (IGF) binding protein-3 (IGFBP-3)는 혈액 내 IGFs와 결합하여 활성도를 조절하는 역할을 한다. 혈액 내 IGFs 및 IGFBP-3의 농도는 개인별로 다양하며 질병에 따라 영향을 받는다. 본 연구에서는 IGFBP-3 유전자형에 따른 신체 지표 및 질병과의 관계를 알아보고자 한다. 방법: 국민건강보험공단에서 일반인을 대상으로 시행하는 검진을 위해 내원한 사람 중에 60세 이상 100명을 무작위로 선택하여 체중, 신장 등의 신체 계측 및 생화학적 검사를 시행하였으며 문진을 통한 과거력, 약물 복용력, 가족력을 조사하였고 말초 혈액에서 DNA를 추출하여 IGFBP-3 promoter -202 locus A/C polymorphism을 검사하였다. 결과: IGFBP-3 promoter -202 locus의 유전자형의 빈도수는 AA 43명(43%), AC 48명(48%), CC 8명(8%)이었다. 통계학적으로 유의 수준에 도달하지는 못하였지만 남녀 모두에서 CC 유전자군의 신장이 가장 큰 반면에 AC 유전자군이 체중, BMI, BSA가 가장 컸다. 여자에서 유전자형에 따른 제2형 당뇨병의 유병률 및 공복 시 혈당에 차이가 없었지만 남자에서는 AC 유전자군이 제2형 당뇨병의 유병률 및 공복 시 혈당이 높게 나왔다(P=0.016, P=0.106). 결론: IGFBP-3의 혈청 농도 및 IGFs의 활성도를 결정하는 IGFBP-3 promoter -202 locus A/C polymorphism은 남자의 경우 제2형 당뇨병의 유병률과 관계가 있으며 이는 IGF BP-3가 혈당의 항상성 유지에 중요한 역할을 하는 것으로 생각된다. Background: Insulin-like growth factor (IGF) binding protein-3 (IGFBP-3) is a major determinant of circulating levels of the IGFs and is clinically useful for the evaluation of GH deficiency and for predicting the response to GH treatment. Circulating levels of IGF-1 and IGFBP-3 vary considerably among individuals and have been implicated in various medical conditions. We investigated the IGFBP-3 genotype distribution in Korean elderly and relations between IGFBP-3 promoter -202 locus A/C polymorphism and medical conditions. Methods: Single nucleotide polymorphism at the -202 locus of the IGFBP-3 promoter was genotyped for 100 samples from a study population who is older than 60 years. Results: The frequency of each polymorphic variation at the -202 locus in these study population was AA=43 (43%), AC=48 (48%), and CC=8 (8%). In both genders, the subjects with the CC genotype were taller, whereas those with AC genotype had higher weight, BMI and BSA comparing those with the other genotypes, but these facts did not reach statistical significance. The prevalence of type 2 DM and the mean fasting blood glucose level did not differ among genotypes in the female gender. In the male population, however, the prevalence of type 2 DM was higher in the subjects with the AC genotype (P=0.016) which is reflected by the elevated mean fasting glucose level (P=0.106). Conclusion: IGFBP-3 promoter -202 locus A/C polymorphism, which reflects its serum level and further implicates IGFs bioactivity, was associated with a higher prevalence of type 2 DM in the male gender, suggesting IGFBP-3 might play a role in blood glucose homeostasis.

자궁내 성장 지연증 소아에서 혈청 IGF-I, Free IGF-I, IGFBP-1, IGFBP-3 농도에 대한 연구

황일태,박은애,김경희,김호성 대한소아내분비학회 1999 Annals of Pediatirc Endocrinology & Metabolism Vol.4 No.2

Identification of Novel SNPs in Bovine Insulin-like Growth Factor Binding Protein-3 (IGFBP3) Gene

Kim, J.Y.,Yoon, D.H.,Park, B.L.,Kim, L.H.,Na, K.J.,Choi, J.G.,Cho, C.Y.,Lee, H.K.,Chung, E.R.,Sang, B.C.,Cheong, I.J.,Oh, S.J.,Shin, Hyoung Doo Asian Australasian Association of Animal Productio 2005 Animal Bioscience Vol.18 No.1

The insulin-like growth factors (IGFs), their receptors, and their binding proteins play key roles in regulating cell proliferation and apoptosis. Insulin-like growth factor binding protein-3 (IGFBP3, OMIM #146732) is one of the proteins that bind to the IGFs. IGFBP3 is a modulator of IGF bioactivity, and direct growth inhibitor in the extravascular tissue compartment. We identified twenty-two novel single nucleotide polymorphisms (SNPs) in IGFBP3 gene in Korean cattle (Hanwoo, Bos taurus coreanae) by direct sequencing of full gene including -1,500 bp promoter region. Among the identified SNPs, five common SNPs were screened in 650 Korean cattle; one SNP in promoter (IGFBP3 G-854C), one in 5'UTR region (IGFBP3 G-100A), two in intron 1 (IGFBP3 G+421T, IGFBP3 T+1636A), and one in intron 2 (IGFBP3 C+3863A). The frequencies of each SNP were 0.357 (IGFBP3 G-854C), 0.472 (IGFBP3 G-100A), 0.418 (IGFBP3 G+421T), 0.363 (IGFBP3 T+1636A) and 0.226 (IGFBP3 C+3863A), respectively. Haplotypes and their frequencies were estimated by EM algorithm. Six haplotypes were constructed with five SNPs and linkage disequilibrium coefficients (|D'|) between SNP pairs were also calculated. The information on SNPs and haplotypes in IGFBP3 gene could be useful for genetic studies of this gene.

Interaction Between Acid-Labile Subunit and Insulin-like Growth Factor Binding Protein 3 Expressed in Xenopus Oocytes

Choi, Kyung-Yi,Lee, Dong-Hee Korean Society for Biochemistry and Molecular Biol 2002 Journal of biochemistry and molecular biology Vol.35 No.2

The acid-bible subunit (ALS) associates with the insulinlike growth factor (IGF)-I or II, and the IGF binding protein-3 (IGFBP-3) in order to form a 150-kD complex in the circulation. This complex may regulate the serum IGFs by restricting them in the vascular system and promoting their endocrine actions. Little is known about how ALS binds to IGFBP3, which connects the IGFs to ALS. Xenopus oocyte was utilized to study the function of ALS in assembling IGFs into the ternary complexes. Xenopus oocyte was shown to correctly translate in vitro transcribed mRNAs of ALS and IGFBP3. IGFBP3 and ALS mRNAs were injected in a mixture, and their products were immunoprecipitated by antisera against ALS and IGFBP3. Contrary to traditional reports that ALS interacts only with IGF-bound IGFBP3, this study shows that ALS is capable of forming a binary complex with IGFBP3 in the absence of IGF When cross-linked by disuccinimidyl suberate, the band that represents the ALS-IGFBP3 complex was evident on the PAGE. IGFBP3 movement was monitored according to the distribution between the hemispheres. Following a localized translation in the vegetal hemisphere, IGFBP3 remained in the vegetal half in the presence of ALS. However, the mutant IGFBP3 freely diffused into the animal half, despite the presence of ALS, which is different from the wild type IGFBP3. This study, therefore, suggests that ALS may play an important role in sequestering IGFBP3 polypeptides via the intermolecular aggregation. Studies using this heterologous model will lead to a better understanding of the IGFBP3 and ALS that assemble into the ternary structure and circulate the IGF system.

유방암에서 Insulin-like Growth Factor Binding Protein (IGFBP)-3 및 PTEN 발현의 감소와 임상병리학적 소견과의 관련성

정성후(Sung Hoo Jung),윤현조(Hyun Jo Youn),김민선(Min Sun Kim),김선영(Sun-Young Kim),황평한(Pyoung Han Hwang),이대열(Dae-Youl Lee) 대한외과학회 2010 Annals of Surgical Treatment and Research(ASRT) Vol.79 No.2

Purpose: Insulin-like growth factor binding protein (IGFBP-3) and phosphatase and tensin homolog (PTEN) are tumor-suppressor genes that may be involved in breast tumorigenesis. However, the roles of these genes in the regulation of breast cancer growth or progress are unclear. In this study, we aimed to find any correlation between the reduction of IGFBP-3 or PTEN protein expression in cancer tissues and the clinicopathological parameters in breast cancer. Methods: We collected both cancer and adjacent normal tissues from 46 breast cancer patients (from January 1 to December 31, 2006), and checked the IGFBP-3 and PTEN protein levels in cancer and adjacent normal tissues using Western immunoblot. We evaluated the correlation of reduction status of IGFBP-3 and PTEN protein expression with variable clinicopathological parameters. Results: The frequency of IGFBP-3 and PTEN protein reduction in cancer tissue, compared to adjacent normal tissue, was 63.0% and 34.8%, respectively. And in 87.5% of patients, who showed significant PTEN reduction, IGFBP-3 protein expression was reduced in cancer tissues. In contrast, IGFBP-3 protein reduced in only 50% of patients who didn’t show PTEN reduction. However, we did not find any significant correlation between reduction of IGFBP-3 or PTEN expression in cancer tissue and variable clinicopathological parameters. Conclusion: The IGFBP-3 and PTEN genes were expressed in all breast cancer tissues. Nonetheless, we did not find any significant relationship between reduction of IGFBP-3 or PTEN expression and the clinicopathological parameters in this study. Therefore, further studies are needed to document the roles of IGFBP-3 and PTEN genes in breast cancer growth or progress.

경증 천식 환아에서 혈중 IGF-I 및 IGFBP-3 농도를 이용한 성장 상태 평가

노창수,김정은,홍성진,정소정,김교순,김덕희 대한소아내분비학회 2004 Annals of Pediatirc Endocrinology & Metabolism Vol.9 No.1

Regulation of replicative senescence by insulin-like growth factor-binding protein 3 in human umbilical vein endothelial cells

Kim, Kwang Seok,Kim, Min-Sun,Seu, Young Bae,Chung, Hae Young,Kim, Jung Hye,Kim, Jae-Ryong BLACKWELL PUBLISHING 2007 AGING CELL Vol.6 No.4

<P>Summary</P><P>Insulin/insulin-like growth factor (IGF) signaling pathways are among the most conserved processes in aging in organisms ranging from yeast to mammals. Previously, using cDNA microarray technology, we reported that expression of IGF-binding protein 3 (IGFBP3), one of the IGF-binding proteins, was increased with age in human dermal fibroblasts. In this study, the role of IGFBP3 on cellular senescence was studied in human umbilical vein endothelial cells (HUVEC). The expression levels of IGFBP3 mRNA and protein were increased in HUVECs with age. Knockdown of IGFBP3 in old cells with IGFBP3 short hairpin RNA (shRNA) retrovirus resulted in the partial reduction of a variety of senescent phenotypes, such as changes in cell morphology, and decreases in population doubling times and senescence-associated &bgr;-galactosidase (SA-&bgr;-gal) staining. Down-regulation of IGFBP3 rescued the growth arrest induced by p53 overexpression in young HUVECs. In contrast, up-regulation of IGFBP3 in young cells and prolonged IGFBP3 treatment accelerated cellular senescence, confirmed by cell proliferation and SA-&bgr;-gal staining. The FOXO3a (forkhead box O3a) protein level was increased in old IGFBP3 shRNA cells. The treatment of young HUVECs with IGFBP3 repressed the levels of FOXO3a protein. Furthermore, calorie restriction reduced IGFBP3 protein levels, which were found to be increased with age in the rat liver and serum. These results suggest that IGFBP3 might play an important role in the cellular senescence of HUVECs as well as <I>in vivo</I> aging.</P>

Hypoxia Suppresses Spontaneous Mineralization and Osteogenic Differentiation of Mesenchymal Stem Cells via IGFBP3 Up-Regulation

Kim, Ji Hye,Yoon, Sei Mee,Song, Sun U.,Park, Sang Gyu,Kim, Won-Serk,Park, In Guk,Lee, Jinu,Sung, Jong-Hyuk MDPI 2016 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.17 No.9

<P>Hypoxia has diverse stimulatory effects on human adipose-derived stem cells (ASCs). In the present study, we investigated whether hypoxic culture conditions (2% O<SUB>2</SUB>) suppress spontaneous mineralization and osteogenic differentiation of ASCs. We also investigated signaling pathways and molecular mechanisms involved in this process. We found that hypoxia suppressed spontaneous mineralization and osteogenic differentiation of ASCs, and up-regulated mRNA and protein expression of Insulin-like growth factor binding proteins (IGFBPs) in ASCs. Although treatment with recombinant IGFBPs did not affect osteogenic differentiation of ASCs, siRNA-mediated inhibition of IGFBP3 attenuated hypoxia-suppressed osteogenic differentiation of ASCs. In contrast, overexpression of IGFBP3 via lentiviral vectors inhibited ASC osteogenic differentiation. These results indicate that hypoxia suppresses spontaneous mineralization and osteogenic differentiation of ASCs via intracellular IGFBP3 up-regulation. We determined that reactive oxygen species (ROS) generation followed by activation of the MAPK and PI3K/Akt pathways play pivotal roles in IGFBP3 expression under hypoxia. For example, ROS scavengers and inhibitors for MAPK and PI3K/Akt pathways attenuated the hypoxia-induced IGFBP3 expression. Inhibition of Elk1 and NF-κB through siRNA transfection also led to down-regulation of IGFBP3 mRNA expression. We next addressed the proliferative potential of ASCs with overexpressed IGFBP3, but IGFBP3 overexpression reduced the proliferation of ASCs. In addition, hypoxia reduced the osteogenic differentiation of bone marrow-derived clonal mesenchymal stem cells. Collectively, our results indicate that hypoxia suppresses the osteogenic differentiation of mesenchymal stem cells via IGFBP3 up-regulation.</P>

사람의 섬유아세포에서 glucose 농도가 물질대사 및 Insulin-like growth factor binding protein-3의 발현에 미치는 영향

류혜영,황혜정,김인혜,류홍수,남택정,Ryu, Hye-Young,Hwang, Hye-Jung,Kim, In-Hye,Ryu, Hong-Soo,Nam, Taek-Jeong 한국생명과학회 2007 생명과학회지 Vol.17 No.5

사람의 섬유아세포인 GM10을 사용하여 glucose의 배양조건에 다른 물질대사 및 IGFBP-3 발현을 살펴본 결과, glucose 농도에 따른 glucose 소비와 triglyceride 축적 수준은 고농도 glucose 배양 조건에서 증가한 반면, 총 단백질 함량은 고농도 glucose배양 조건에서 시간의 경과에 따라 감소하였다. 또한 고농도 glucose 배양 조건에서 5일 동안 배양한 세포 배양액내의 유리아미노산 함량은 저농도보다 고농도 glucose 배양 조건에서 높게 나타났다. IGFBP-3 단백질 수준과 mRNA수준은 저농도 glucose배양 조건에서 증가하였으나, IGFBP-3 단백질 분해효소에 따른 영향은 없었다. 이상의 결과에서 glucose 배양조건에 따른 물질대사는 부분적으로 세포를 이용한 당뇨병 모델 실험에서 in vivo와 같은 결과를 얻지 못하였지만, IGFs와 같은 세포 성장인자에 대한 연구를 통해 세포수준의 당뇨병 모델화가 가능하다고 여겨진다. Insulin-like growth factor-I(IGF-I) has significant insulin-like anabolic effects which include the stimulation of glucose and amino acid uptake, as well as protein and glycogen synthesis. IGFs exist in serum and other biological fluids as complexes bound to a family of structurally related insulin-like growth factor binding proteins(IGFBPs). Six human IGFBPs can modulate the effects of IGFs on target tissues by several mechanisms, including altering the serum's half-life and the transcapillary transport of IGFs, as well as changing the availability of IGFs to specific cell surface receptors. Human fibroblasts secrete IGFBPs that can modify IGF-I action. Previous to our study using either Northern blotting, and Western blotting have shown that fibroblasts express mRNA IGFBP-3, -4, and -5, and synthesize these proteins. In addition, fibroblast cell lysates revealed that the IGFBP-3 was most abundant. For these reasons, we undertook to gain further insight into the effects of high and low glucose incubation condition on metabolism and IGFBP-3 expression. In results of metabolites and IGFBP-3 expression in GM10 cells cultivated with various glucose concentration, the consumption of glucose and accumulation of triglyceride were increased in condition of high glucose, and total protein level was decreased. in the course of time. After 5 days incubation, levels of free amino acid in medium containing glucose of high concentration glucose were higher than in conditions of low glucose. Although the levels of IGFBP-3 protein and mRNA levels were increased in low glucose, and IGFBP-3 was not affected by any pretense. Taken together, we suggest that the study of growth factors, like IGFs, might be a possible model of diabetes militus in cell, although the results in cell models were not in accord with in vivo.

Effects of Volatile Fatty Acids on IGF-I, IGFBP-3, GH, Insulin and Glucagon in Plasma, and IGF-I and IGFBP-3 in Different Tissues of Growing Sheep Nourished by Total Intragastric Infusions

Zhao, Guang-Yong,Sun, Ya-Bo Asian Australasian Association of Animal Productio 2010 Animal Bioscience Vol.23 No.3

Twelve Suffolk${\times}$Small-tail-Han male sheep (body weight 21-26 kg), aged four months, were used to study the effects of volatile fatty acids (VFA) on IGF-I (insulin-like growth factor-I), IGFBP-3 (insulin-like growth factor binding protein-3), GH (growth hormone), insulin and glucagon in plasma, and IGF-I and IGFBP-3 in different tissues. The sheep were randomly divided into four groups with 3 sheep in each group. The sheep were sustained by total intragastric infusions and four levels of mixed VFA (the molar proportion of acetic acid, propionic acid and butyric acid was 65:25:10), which supplied 333, 378, 423 and 468 KJ energy/kg $W^{0.75}$/d, were infused into the rumen as experimental Treatments I, II, III and IV, respectively. The experiment lasted 12 days, of which the first 8 days were for pretreatment and the last 4 days for collection of samples. At the end of the experiment, blood samples were taken and then the sheep were slaughtered and tissue samples from the rumen ventral sac, rumen dorsal sac, liver, duodenum and Longissimus dorsi muscle were obtained. IGF-I, IGFBP-3, GH, insulin and glucagon in plasma and IGF-I and IGFBP-3 in different tissues were analysed. Results showed that the concentration of IGF-I, IGFBP-3, GH, insulin or glucagon in plasma and the content of IGF-I and IGFBP-3 in the rumen dorsal sac, rumen ventral sac, liver or Longissimus dorsi muscle were increased with VFA infusion level (p<0.05). No significant differences were found in duodenum IGF-I between Treatments I and II and in rumen dorsal sac IGFBP-3 between Treatments II and III (p>0.05). It was concluded that IGF-I, IGFBP-3, GH, insulin and glucagon in plasma and IGF-I and IGFBP-3 in rumen dorsal sac, rumen ventral sac, liver and Longissimus dorsi muscle were increased significantly with increasing level of ruminal infusion of mixed VFA.