• Asian-Australasian Journal of Animal Sciences
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• 제18권1호
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• pp.3-7
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• 2005

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.

• Asian-Australasian Journal of Animal Sciences
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• 제25권11호
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• pp.1515-1520
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• 2012

Insulin-like growth factor binding protein-3 (IGFBP-3) gene is important for regulation of growth and development in mammals. The present investigation was carried out to study DNA polymorphism by PCR-RFLP of IGFBP-3 gene and its effect on fibre traits of Chinese Inner Mongolian cashmere goats. The fibre traits data investigated were cashmere fibre diameter, combed cashmere weight, cashmere fibre length and guard hair length. Four hundred and forty-four animals were used to detect polymorphisms in the hircine IGFBP-3 gene. A 316-bp fragment of the IGFBP-3 gene in exon 2 was amplified and digested with HaeIII restriction enzyme. Three patterns of restriction fragments were observed in the populations. The frequency of AA, AB and BB genotypes was 0.58, 0.33 and 0.09 respectively. The allelic frequency of the A and B allele was 0.75 and 0.25 respectively. Nucleotide sequencing revealed a C>G transition in the exon 2 region of the IGFBP-3 gene resulting in R158G change which caused the polymorphism. Least squares analysis revealed a significant effect of genotypes on cashmere weight (p<0.0001), cashmere fibre length (p<0.001) and hair length (p<0.05) of the animals. The effect of genotypes on cashmere fibre diameter was not statistically significant (p>0.05). The animals of AB and BB genotypes showed higher cashmere weight, cashmere fibre length and hair length than the animals possessing AA genotype. These results suggested that polymorphisms in the hircine IGFBP-3 gene might be a potential molecular marker for cashmere weight in cashmere goats.

• Asian-Australasian Journal of Animal Sciences
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• 제17권7호
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• pp.910-913
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• 2004

Insulin-like growth factor binding protein-3 (IGFBP-3) gene is a structural gene associated with the growth and development of the animals. The present investigation was carried out to unravel nucleotide sequence and polymerase chain reactionrestriction fragment polymorphism (PCR-RFLP) of IGFBP-3 gene in buffalo. Genomic DNA was isolated from a total of 157 animals belonging to Murrah, Surti, Jaffarabadi and Nagpuri breeds of Indian riverine buffalo. A 655 bp of IGFBP-3 gene was amplified in all the breeds and amplicons were digested with Hae III, Taq I and Msp I restriction enzymes. On digestion with Hae III yielded single restriction pattern of 8 fragments of sizes 201, 165, 154, 56, 36, 19, 16 and 8 bp in all the animals studied. Similarly Taq I and Msp I also revealed single restriction pattern yielding fragments of sizes 240 and 415 bp and 145 and 510 bp, respectively. This shows nonpolymorphic nature of restriction sites in buffalo. Nucleotide sequencing of 587 bp of IGFBP-3 gene in Murrah buffalo was done and submitted to the GenBank (Accession No. AY304829). Nucleotide sequencing revealed an addition of 4 bases in the intronic region as compared to cattle.

• Clinical and Experimental Pediatrics
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• 제49권4호
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• pp.431-438
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• 2006

목 적 : 새로운 암억제 유전자로 알려진 IGFBP-3의 주요 기능은 IGF-I과 IGF-II와 결합을 하여 IGF의 기능을 조절하는 IGF dependent mechanism과 IGFBP-3 자체가 IGF와 결합과는 무관하게 세포의 apoptosis를 유도하는 IGF independent mechanism이 보고되었다. 암억제 유전자 p53의 대표적인 항암 기전의 하나는 직접 IGFBP-3의 발현을 증가시키며, 발현된 IGFBP-3는 암세포의 apoptosis를 유도시킨다. IGFBP-3의 항암작용은 보고되었지만, IGFBP-3의 변역 후 변형에 의한 항암기전은 전혀 밝혀져 있지 않다. 본 연구에서는 p53의 항암기전과 관련하여 IGFBP-3의 당화에 관련된 기전을 밝히고, IGFBP-3 당화의 의미를 규명하였다. 방 법 : 실험 세포주로는 p53의 변이를 보이며 p53의 발현이 일반세포에 비교하여 낮은 특징을 갖고 있는 사람의 유방암세포인 MDA-MB-231를 사용하였으며, Ad/p53과 Ad/IGFBP-3 아데노바이러스를 감염시킨 후 IGFBP-3의 발현 변화와 apoptosis 기전을 분석하였다. glycosylation 억제제로 알려져 있는 tunicamycin을 처리하여 당화의 정도를 조사하였다. 결 과 : 실험 세포에 Ad/p53을 감염시켜 p53을 발현시킨 결과 성장의 억제와 apoptosis가 유도되었고, IGFBP-3의 발현이 현저하게 증가되었으며, 특히 IGFBP-3의 당화 형태를 증가시켰다. 당화된 IGFBP-3의 증가는 세포의 apoptosis의 유도가 촉진되었으며, 이러한 IGFBP-3의 당화는 p53과 IGFBP-3의 발현을 동시에 유도시킨 결과 더욱 항진되었다. 결 론 : 이상의 연구에서 IGFBP-3의 암억제 능력의 향상은 p53에 의한 IGFBP-3의 당화의 증가를 통하여 안정화됨으로서 나타나고 있음을 알 수 있었다. 이는 p53과 IGFBP-3를 이용한 혼합유전자 치료가 가능할 것으로 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제21권3호
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• pp.309-313
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• 2008

The insulin-like growth factor binding protein 3 (IGFBP3) has been investigated as a candidate gene for growth promoting effects in beef cattle and a modulator of IGF bioactivity. Previously, we have reported twenty two sequence variants discovered in Korean native cattle (Hanwoo). In this study, we examined the association between gene-specific polymorphisms of IGFBP3 and cold carcass weight (CW) and marbling score (MS) among Korean native cattle. Among twenty two polymorphisms, four common polymorphic sites (-854G>C, -100G>A, +421G>T and +3863C>A) were genotyped in our beef cattle (n = 437). Statistical analysis revealed that one common polymorphism in the promoter region (-854G>C) showed putative associations with MS (p = 0.03). IGFBP3 variation/haplotype information analyzed in this study will provide valuable information into strategies for the production of a commercial line of beef cattle.

• Asian Pacific Journal of Cancer Prevention
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• 제15권21호
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• pp.9327-9333
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• 2014

Breast cancer is the second most common cancer and second leading cause of cancer deaths in women. Phosphatidylinositol-3-kinase (PI3K)/AKT pathway mutations are associated with cancer and phosphatidylinositol-4, 5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) gene mutations have been observed in 25-45% of breast cancer samples. Insulin growth factor binding protein-5 (IGFBP-5) can show different effects on apoptosis, cell motility and survival in breast cancer. We here aimed to determine the association between PIK3CA gene mutations and IGFBP-5 expressions for the first time in breast cancer patients. Frozen tumor samples from 101 Turkish breast cancer patients were analyzed with high resolution melting (HRM) for PIK3CA mutations (exon 9 and exon 20) and 37 HRM positive tumor samples were analyzed by DNA sequencing, mutations being found in 31. PIK3CA exon 9 mutations (Q546R, E542Q, E545K, E542K and E545D) were found in 10 tumor samples, exon 20 mutations (H1047L, H1047R, T1025T and G1049R) in 21, where only 1 tumor sample had two exon 20 mutations (T1025T and H1047R). Moreover, we detected one sample with both exon 9 (E542Q) and exon 20 (H1047R) mutations. 35% of the tumor samples with high IGFBP-5 mRNA expression and 29.4% of the tumor samples with low IGFBP-5 mRNA expression had PIK3CA mutations (p=0.9924). This is the first study of PIK3CA mutation screening results in Turkish breast cancer population using HRM analysis. This approach appears to be a very effective and reliable screening method for the PIK3CA exon 9 and 20 mutation detection. Further analysis with a greater number of samples is needed to clarify association between PIK3CA gene mutations and IGFBP-5 mRNA expression, and also clinical outcome in breast cancer patients.

• Asian-Australasian Journal of Animal Sciences
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• 제22권4호
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• pp.471-482
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• 2009

Nutrient availability may control muscle growth directly and indirectly through its influence on regulatory factors. We analyzed the effects of nutrient availability on the breast muscle insulin-like growth factor system. Real time RT-PCR was used to quantify the level of transcription in breast muscle from Langshan (LS) layer and Arbor Acres (AA) broiler chickens subjected to different feeding regimens during embryonic and postnatal development. The AA chickens were fed AA diet (AA, control group) while the LS chickens were either fed LS diet (LL) or AA diet (LA). According to our results, insulin-like growth factor (IGF)-II (embryonic day 16 (E16) - postnatal day 42 (P42)), IGF-I receptor (IGF-IR, E18-P42), and IGF binding protein (IGFBP)-2 (E18-P42), -5 (E16-P14), -7 (E12-P0), and -3 (E12-P0) were positively correlated with IGF-I, while IGFBP-3 (P0-P28) was negatively correlated with IGF-I. In comparison, IGF-IR (E18-P42), IGFBP-2 (E18-P42), IGFBP-5 (E14-P0), and IGFBP-3 (E16-P0) were positively correlated with IGF-II, while IGF-IR (E10-E16) and IGFBP-3 (P0-P28) were negatively correlated with IGF-II. Moreover, IGFBP-2 (E16-P42), -7 (E10-E16), and -3 (E10-E16) were positively correlated with IGF-IR, while IGFBP-3 (P0-P28) was negatively correlated with IGF-IR. Finally, IGFBP-7 (E12-P0) was positively correlated with IGFBP-3, while IGFBP-2 (P0-P28) and -7 (P0-P42) were negatively correlated with IGFBP-3. Overall, the AA chickens exhibited higher levels of IGF-I, IGF-IR, and IGFBP-2 mRNA expression than the LL chickens, while the opposite was true for IGFBP-7. No strain differences in IGF-I, IGF-IR, and IGFBP-7 mRNA expression were detected between LA and AA chickens; however, a strain difference was observed for IGFBP-2. LA chickens exhibited higher levels of IGFBP-2 than LL chickens, while the opposite was true for IGFBP-7. Our data show the first evidence that certain genes may be correlated during specific developmental periods and that strain differences in the expression of those genes in LS and AA chickens are due to differential responses to the same diet.

• Asian-Australasian Journal of Animal Sciences
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• 제19권8호
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• pp.1095-1099
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• 2006

Leptin and IGFBP-3 are two proteins that play an important role in growth and metabolism of the animals. They are also involved in the immune function of animals and, thus, are candidate genes for the study of association with immune functions. Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) of these two genes was done to screen 64 crossbred (Holstein Friesian${\times}$Hariana) female calves of one year of age. From each RFLPs (fragments) three genotypes were observed. In all the RFLPs the mutant homozygotes were very less in numbers and, hence, were excluded from the least squares analysis. The serum IgG level was estimated using SRID assay. The mean level of serum IgG was $28.83{\pm}2.73mg/ml$. The effect of these identified genotypes on serum IgG level of calves at one year of age was analysed using least squares analysis. The HaeIII RFLP-AB genotype had significantly (p<0.05) higher serum IgG level ($31.86{\pm}3.05$) than the HaeIII RFLP-AA ($25.62{\pm}2.96$) genotype. There was no significant effect of leptin genotypes on the IgG level. The present results indicated a role of the IGFBP-3 gene on serum IgG level of cattle calves.

• Tuberculosis and Respiratory Diseases
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• 제58권4호
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• pp.359-366
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• 2005

인슐린양 성장 인자 결합 단백-3(Insulin-like growth factor (IGF) binding protein-3 (IGFBP-3))는 혈액 내에서 IGF와 결합하여 복합체 혹은 저장소로 작용함으로써, IGF가 수용체에 결합하는 것을 방해하여 IGF의 항세포사멸(antiapoptosis) 및 세포분열 촉진의 기능을 억제한다. 하지만, 특정 상황에서는 도리어 IGFBP-3가 IGF의 파괴를 억제하여 IGF에 의한 암세포의 분화 및 성장을 촉진할 수도 있다는 것이 알려져 있다. 대부분의 환자에서 혈액내 IGFBP-3 수치는 IGFBP-3 유전자의 -202 좌위(locus)의 다형성(polymorphism)에 의해 크게 영향을 받는다. 따라서, 저자 등은 제한 효소(restriction enzyme)를 이용하여 비소세포폐암 환자의 IGFBP-3 유전자 -202 좌위의 다형성을 분석함으로써, 이 좌위의 다형성이 비소세포폐암의 위험도와 연관되어 있는지 조사하였다. 본 연구는 104명의 비소세포폐암 환자군과, 연령, 성별, 흡연력이 비슷한 104명의 대조군을 비교 분석하였다. 대조군에서 -202 좌위 유전자 다형성의 빈도는 AA형 48명 (46.2%), AC형 45명 (43.3%), CC형 11명 (10.5%)이었고, 비소세포폐암 환자군에서 -202 좌위 유전자 다형성의 빈도는 AA형 67명(64.4%), AC형 35명 (33.7%), CC형 2명 (1.9%)이었다. -202 좌위의 유전자 다형성에 있어서 대조군과 비소세포폐암 환자군 사이에 유의한 빈도 차이가 있었으며 (p < 0.05, Pearson's ${\chi}^2-test$), 비소세포폐암의 위험도는 -202 좌위의 AA형에서 가장 높고 CC형에서 가장 낮았다. CC형을 기준으로 하면 AC형의 비교 위험도는 2.60 (95% 신뢰구간: 0.89 - 8.60)이었으며 AA형의 비교 위험도는 5.89 (95% 신뢰구간: 1.92 - 21.16)이었다. 본 연구 결과는, IGFBP-3 유전자의 -202 좌위(locus)의 다형성(polymorphism)이 비소세포폐암의 위험인자 중의 하나일 가능성을 제시하며, 따라서 비소세포폐암에 대한 항암치료 개발에 있어서 새로운 표적이 될 가능성을 시사한다.

• 대한수의학회지
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• 제46권2호
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• pp.97-105
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• 2006

All-trans retinoic acid (AtRA) induces growth inhibition and apoptosis in a variety of tumer cells, including MCF-7 cells. Insulin-like growth factors (IGFs) system has been reported to be associated with the development of cancer. Although MCF-7 cell with AtRA is to be the major stimulus for the cell growth and apoptosis, the mechanism of insulin-like growth factor-I (IGF-I)/insulin-like growth factor binding protein-3 (IGFBP-3) system remains to be elucidated. Thus, this study was conducted to the effect of AtRA on the gene expression and level of IGF-I and IGFBP-3. In addition, we investigated the involvement of PKC-${\delta}$ on the IGF-I and IGFBP-3 secretion in MCF-7 cell. AtRA(${\geq}10^{-7}M$) decreased the IGF-1 secretion and mRNA expressions, but increased IGFBP-3 secretion and mRNA expressions in MCF-7 cells. Especially, the treatment of AtRA at 72 hours caused a significant reduction in the IGF-I secretion and mRNA expressions but increment in IGFBP-3 secretion and mRNA expressions (p < 0.05). $10^{-7}M$ AtRA activated PKC-${\delta}$ that is one among PKC-$\iota$, ${\alpha}$, ${\lambda}$ and ${\delta}$ in MCF-7 cell. Rotllerin, a PKC-${\delta}$ inhibitor, blocked AtRA-induced inhibition of the IGF-I and mRNA expressions, and increase of lGFBP-3 and mRNA expressions in MCF-7 cell. Together, AtRA inhibited the IGF-I secretion and mRNA expressions, but increased IGFBP-3 secretion and mRNA expressions in MCF-7 cell. Furthermore, AtRA-induced alteration of IGF-I, IGFBP-3 secretion, and the gene expressions were mediated via PKC-${\delta}$ activity.