• Clinical and Experimental Pediatrics
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• 제48권3호
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• pp.298-305
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• 2005

목 적 : 혈청 IGF-I, IGFBP-2, IGFBP-3 농도 및 IGF-I/IGFBP-3 molar ratio는 소아의 성장 발달, 당뇨병, 비만 및 악성종양의 진단, 예후 예측과 치료후 추적에 유용한 지표이지만 성별, 연령별, 인종별 변이가 있다. 저자들은 우리나라 어린이의 성별 및 연령군별 혈청 IGF-I, IGFBP-2, IGFBP-3 농도 및 IGF-I/IGFBP-3 molar ratio의 정상 표준치 확립을 위하여 본 연구를 시행하였다. 방 법 : 총 92명(남자 42명, 여자 50명)의 건강한 어린이를 대상으로 하였고, 신생아군 9명(남자 3명, 여자 6명), 영아군 19명(남자 9명, 여자 10명), 유아군 24명(남자 13명, 여자 11명), 학령군 30명(남자 12명, 여자 18명), 청소년군 10명(남자 5명, 여자 5명)으로 분류 후, ELISA 방법을 이용하여 혈청 IGF-I, IGFBP-2 및 IGFBP-3 농도를 측정하였고, 각 군간의 IGF-I, IGFBP-2, IGFBP-3 및 IGF-I/IGFBP-3 molar ratio를 비교분석하였다. 결 과 : 1) 남자에서는 IGF-I과 IGFBP-3는 출생과 더불어 증가하여 각각 유아군(P<0.05), 학령군(P<0.05)에 최고값에 도달한 후 청소년군으로 갈수록 감소하였다. IGFBP-2와 IGF-I/IGFBP-3 molar ratio는 각각 학령군과 영아군에 최고값을 보이고 감소하였으나 출생시에 비해 통계적으로 유의한 증가를 보이지 않았다. 각 군간의 비교에서 IGFBP-2만이 유의한 차이를 보였다(P<0.05). 2) 여자에서는 IGF-I과 IGFBP-2, IGFBP-3는 출생과 더불어 증가하여 각각 청소년군, 학령군(P<0.05), 학령군(P<0.05)에 최고값에 도달하였으나 IGF-I은 출생시에 비해 통계적으로 유의한 증가를 보이지 않았다. IGF-I/IGFBP-3 molar ratio는 출생시에 최고값을 보였으나 통계적으로 유의한 차이를 보이지 않았다. 각 군간의 비교에서 IGFBP-2 및 IGFBP-3가 유의한 차이를 보였다(P<0.05). 3) 동일연령군별 남녀 비교에서 IGF-I과 IGFBP-2, IGFBP-3 및 IGF-I/IGFBP-3 molar ratio 어느 것도 남녀 간에 통계적으로 유의한 차이를 보이지 않았다. 4) 상관관계 조사시 IGF-I의 농도가 높을수록 IGFBP-3의 농도 역시 의미있게 증가하였고(r=0.504, P=0.000), IGF-I/IGFBP-3 molar ratio와 비만도와의 상관 관계를 조사해 본 결과 통계적으로 의미있는 음의 상관 관계를 보이지 않았다(r=0.051, P=0.629). 또한 비만도와 IGF-I, IGFBP-2 및 IGFBP-3 사이에 통계적으로 유의한 상관 관계는 없었다. 결 론 : IGF-I과 IGFBP-3는 사춘기 진행에 따라 증가하며, 본 연구에서 IGFBP-3가 학령군에서 최고 수준을 보이는 것은 사춘기가 일찍 오는 요즘 어린이들의 성장 추세를 반영하는 것으로 추정되어진다. IGFBP-2는 신생아군에서 가장 높고 영아군에서 감소하는 데 이것은 IGFBP-2가 출생 전 발달 과정에 중요한 물질임을 뒷받침해주는 소견이다. 향후 더 많은 정상아를 대상으로한 연구를 통하여 보다 정확한 우리나라 어린이의 성별 연령군별 정상 표준치 확립이 필요하겠다.

• Asian-Australasian Journal of Animal Sciences
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• 제14권12호
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• pp.1781-1784
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• 2001

We have examined the influence of antisense oligo deoxynucleotide (ODN) of IGFBP-2 on tissue IGFBP-2 gene expression in chicks. Antisense IGFBP-2 ODN was directly injected into the liver or cerebroventricle. Control birds were injected with vehicle. The hepatic IGFBP-2 gene expression was decreased to approximately 30% of the control at 2 h after injection of antisense ODN. In the brain of chickens injected with antisense ODN, IGFBP-2 mRNA level did not change after 2 h of injection and decreased to approximately 60% of the control after 6 h of injection. These results showed that the expression of IGFBP-2 gene in the liver and brain was successfully suppressed by administrating antisense ODN and that hepatic IGFBP-2 gene expression was quickly suppressed by antisense ODN compared with the brain.

• Asian-Australasian Journal of Animal Sciences
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• 제18권11호
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• pp.1575-1581
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• 2005

This study was conducted to determine the characteristics of IGFBPs in plasma of steers, and to profile the relationship between growth and plasma IGF-1 and IGFBPs with aging in Holstein steers. Four blots of IGFBP at molecular weights of 38-43, 34, 29-32 and 24 kDa bands were detected by western ligand blot assay using $^{125}I-IGF-1$. On the basis of immunoblotting with anti-bovine IGFBP-2 and -3 antiserums, we observed the band for IGFBP-2 at approximately 34 kDa, and the IGFBP-3 band was detected at 38-43 kDa and 34 kDa in adult steers and calves. The IGFBP-3 antiserum used on the blots exhibited significant cross-reactivity with 34 kDa IGFBP-2. Furthermore, the 38-43 kDa IGFBP-3 bands were reduced to a 36 kDa band after deglycosylation, whereas the 34 kDa IGFBP-2 was intact. The plasma IGF-1, IGFBP-3 and other IGFBPs showed stability throughout a whole day. The change in live weight was found to be positively correlated to the plasma IGF-1 concentration (r = 0.6801, n = 64, p<0.05) and plasma IGFBP-3 (r = 0.6321, n = 64, p<0.05), while inversely correlated to plasma IGFBP-2 (r = -0.2919, n = 64, p<0.05). Furthermore, plasma IGF-1 was positively correlated to plasma IGFBP-3 (r = 0.6191, p<0.001), but was not correlated to plasma IGFBP-2. The portion of IGFBP-2 for total IGFBPs in calves was higher than in adult steers (p<0.05) and was decreased with growth, whereas that of IGFBP-3 was increased with increased live weight (p<0.05). The ratio IGFBP-3 for IGFBP-2 (BP-3/BP-2) was increased with growing of liveweight. Therefore, the changes in plasma IGF-1 level with increased liveweight may be related to the changes in plasma IGFBP-3 level and IGFBP-2 may give an important role in anabolic action of IGF-1 with the growth of body during calfhood in Holstein steers.

• Childhood Kidney Diseases
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• 제3권2호
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• pp.109-116
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• 1999

목적 : Insuln-like growth factor(IGF)-I및 -II는 성장인자로 일군의 IGF-binding protein(IGFBP)에 의하여 그 작용이 조절된다. IGF-I은 백서 신장에서 발견되고 대사효과와 성장효과를 갖고 있다. 이번 연구는 백서에서 신장발육과 허혈성 신손상 후 재생과정 동안에 IGFBP의 발현이 변화하는지를 보고자 한다. 방법 : 태생 15주부터 성숙 할때까지 백서 신장에서 IGFBP 발현의 변화를 알아보기 위해 Northern blotting을 시행하였고, 급성신부전 백서의 신장에서 IGF-IGBP axis의 변화를 보기 위해 Northern blotting과 Immunohisto-chemistry를 이용하였다. 결과 : IGFBP-1과 -3는 태생기에는 거의 발현되지 않다가 출생 후 7일째부터 성숙이 끝날 때까지 점진적으로 증가하였다. 반면에 IGFBP-2와 -5는 태생기에 많이 발현되고, IGFBP-2는 출생 후 7일째까지 IGFBP-5는 30일째까지 높은 농도를 유지하다가 급격히 감소하였다. 한편 IGFBP-4는 태생기에 중등도로 발현되는데 출생 후 증가하기 시작하여 7일째 가장 많이 발현되다가 급격히 감소하였다. 신손상 후 IGFBP의 변화를 보면 IGFBP-1과 -4는 재생기간 동안 3-5배 증가되다가 정상으로 회복된 반면 IGFBP-3와 IGFBP Related Protein-1(IGFBPrP-1) 은 신손상 1일째에는 약간 감소하다가 그후 증가하여 정상보다 약간 높은 수준을 유지하였다. 결론 : 백서 신장에서 신장구조나 기능의 발달, 분화 및 재생에 대한 IGF의 작용을 조절할 수 있는 IGFBP 발현에 현저한 변화가 있었다.

• BMB Reports
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• 제32권3호
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• pp.266-272
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• 1999

The insulin-like growth factor (IGF) system consisting of IGF-I, IGF-II, IGF-receptors, and IGF-binding proteins (IGFBP) regulates the proliferation of a variety of cancer cell types. To examine whether a decrease in endogenous IGFBP-3 stimulates proliferation or inhibits differentiation, Caco-2 cells, a human colon adenocarcinoma cell line, were stably transfected with an anti-sense IGFBP-3 expression construct or pcDNA3 vector as control. Accumulation of IGFBP-3 mRNA and secretion of IGFBP-3 into serum-free conditioned medium, 9 days after plating, were significantly lower in Caco-2 cell clones transfected with anti-sense IGFBP-3 cDNA compared to the controls. The anti-sense clones grew at a similar rate to the controls for 8 days after plating, but achieved a higher final density between days 10 and 12. The levels of sucrase-isomaltase mRNA, a marker of enterocyte differentiation of Caco-2 cells, were lower in the anti-sense clones examined on day 9. In conclusion, proliferation of Caco-2 cells can be stimulated by lowering endogenously-produced IGFBP-3.

• 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.

• Tuberculosis and Respiratory Diseases
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• 제56권5호
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• pp.465-484
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• 2004

배 경 : 인슐린 양 성장 인자(IGF) 결합 단백질-3(IGFBP-3)은 IGF와 결합하여 IGF의 세포 분열 촉진 및 항세포 고사 기전을 억제할 뿐 아니라 IGF와는 독립적으로 세포고사를 유도함으로써 비소세포성 폐암 세포주의 성장을 억제한다. 방 법 : 본 연구에서 저자들은 IGFBP-3 promoter의 hyper-methylation이 IGFBP-3 단백 발현에 어떠한 역할을 하는가를 연구하였다. 또한 비소세포성 폐암 세포주에서 methylation된 IGFBP-3 promoter에서 유전자 발현을 억제하는 기전을 연구하였다. 결 과 : 본 연구에 사용된 15 종의 비소세포성 폐암 세포주 중 7종 (46.7%)에서 IGFBP-3 promoter의 methylation 이 관찰되었으며, 23명의 I기 환자 검체 중 16 (69.7%), 9명의 II기 환자 검체중 7 (77.8%), 5명의 IIIA 환자 검체중 4 (80%), 6명의 IIIB 환자 검체중 4 (66.7 %), 그리고 6 명의 IV기 환자검체중 6명 모두에서 (100%) promoter 의 methylation 이 관찰되었다. 이 비소세포성 폐암 세포주에서 promoter methylation 상태는 IGFBP-3 단백 및 mRNA 발현양상과 잘 일치하였으며, IGFBP-3의 발현이 억제되었던 비소세포성 폐암 세포주들 중 일부의 세포에서 demethylating 약제인 5'-aza-2'-deoxycytidine (5'-aza-dC) 처리 후 그 발현이 회복되었다. IGFBP-3 promoter 활성도에 중요한 역할을 하는 Sp-1/Sp-3 결합 요소는 IGFBP-3 단백 발현이 억제된 비소세포성 폐암 세포주에서 methylation되어 있었으며, 이 요소의 methylation 은 Sp-1 전사 인자의 결합을 억제하였다. ChIP assay 결과에서 IGFBP-3 promoter의 methylation 상태는 Sp-1/Sp-3 결합 요소에 Sp-1, methyl-CpG binding protein-2 (MeCP2), 그리고 histone deacetylase (HDAC)의 결합에 영향을 주며, 이는 5'-aza-dC 처리에 의하여 역전 되었다. Sp-1/Sp-3 결합 요소를 포함하고 있는 IGFBP-3 promoter의 in vitro methylation은 promoter activity를 현저히 감소시켰으며 이는 MeCP2 단백을 동시에 발현 시켰을 때 더욱 억제되며 5'-aza-dC 처리시 회복되었다. 결 론 : 이러한 결과들은 IGFBP-3 promoter의 methylation이 IGFBP-3 발현을 억제하는 하나의 기전이며, HDAC의 모집을 유도함으로서 MeCP2가 IGFBP-3 발현 억제에 중요한 역할을 함을 보이는 것이다. 이런 현상은 비소세포성 폐암에서 진단 당시의 진행된 병기와도 관계가 있어 IGFBP-3 promoter의 methylation 상태가 비소세포성 폐암의 발암 기전 및 진행에 중요한 역할을 하고 있음을 보이고 있으며, 나아가 조기 진단 및 암 예방영역에서 하나의 생물학적 지표로도 사용될 수 있을 것으로 생각된다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권23호
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• pp.10085-10089
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• 2015

Background: The insulin-like growth factor (IGF) system comprises a group of proteins that play key roles in regulating cell growth, differentiation, and apoptosis in a variety of cellular systems. The aim of this study was to investigate the role of insulin-like growth factor binding protein 3 (IGFBP3) in hepatocellular carcinoma. Materials and Methods: Expression of IGF2, IGFBP3, and PTEN was analyzed by qRT-PCR. Lentivirus vectors were used to overexpress IGFBP3 in hepatocellular carcinoma cell (HCC) lines. The effect of IGFBP3 on proliferation was investigated by MTT and colony formation assays. Results: Expression of IGF2, IGFBP3, and PTEN in several HCC cell lines was lower than in normal cell lines. After 5-aza-2'-deoxycytidine/trichostatin A treatment, significant demethylation of the promoter region of IGFBP3 was observed in HCC cells. Overexpression of IGFBP3 induced apoptosis and reduced colony formation in HUH7 cells. Conclusions: Expression of IGF2, IGFBP3, and PTEN in several HCC cell lines was lower than in normal cell lines. After 5-aza-2'-deoxycytidine/trichostatin A treatment, significant demethylation of the promoter region of IGFBP3 was observed in HCC cells. Overexpression of IGFBP3 induced apoptosis and reduced colony formation in HUH7 cells.

• BMB Reports
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• 제35권2호
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• pp.186-193
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• 2002

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.

• The Korean Journal of Physiology and Pharmacology
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• 제17권2호
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• pp.157-162
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• 2013

Insulin-like growth factor binding proteins (IGFBPs) are important components of insulin growth factor (IGF) signaling pathways. One of the binding proteins, IGFBP-5, enhances the actions of IGF-1, which include the enhanced proliferation of smooth muscle cells. In the present study, we examined the expression and the biological effects of IGFBP-5 in vascular smooth muscle cells (VSMCs) from spontaneously hypertensive rats (SHR) and Wistar Kyoto rats (WKY). The levels of IGFBP-5 mRNA and protein were found to be higher in the VSMC from SHR than in those from WKY. Treatment with recombinant IGFBP-5-stimulated VSMC proliferation in WKY to the levels observed in SHR. In the VSMCs of WKY, incubation with angiotensin (Ang) II or IGF-1 dose dependently increased IGFBP-5 protein levels. Transfection with IGFBP-5 siRNA reduced VSMC proliferation in SHR to the levels exhibited in WKY. In addition, recombinant IGFBP-5 significantly up-regulated ERK1/2 phosphorylation in the VSMCs of WKY as much as those of SHR. Concurrent treatment with the MEK1/2 inhibitors, PD98059 or U0126 completely inhibited recombinant IGFBP-5-induced VSMC proliferation in WKY, while concurrent treatment with the phosphatidylinositol-3 kinase inhibitor, LY294002, had no effect. Furthermore, knockdown with IGFBP-5 siRNA inhibited ERK1/2 phosphorylation in VSMC of SHR. These results suggest that IGFBP-5 plays a role in the regulation of VSMC proliferation via ERK1/2 MAPK signaling in hypertensive rats.