Growth and Pituitary Hormonal Status in Children with Craniopharyngioma / 대한소아내분비학회지

PURPOSE:Craniopharyngioma is one of the most common causes of organic growth hormone deficiency leading to pituitary hormonal insufficiency. However, some growth hormone(GH)-deficient children with craniopharyngioma may grow normally or even show accelerated growth. This study was designed to evaluate several factors associated with growth of patients with craniopharyngioma. METHODS:Forty children operated on for craniopharyngioma were evaluated for their pituitary function, serum insulin like growth factor-I(IGF-I), serum insulin like growth factor binding protein-3(IGFBP-3) and serum prolactin levels. We also observed their growth status and corresponding changes with or without GH treatment. RESULTS:Among 40 patients, one had normal pituitary hormonal status and one had isolated GHD(GH deficiency). The other patients showed multiple pituitary hormone deficiency including GH(98%), LH, FSH(75%), TSH(65%), ACTH(62%), and ADH(38%) deficiencies. Patients with GHD were categorized into 2 groups. Group 1 consisted of children who showed normal growth, thus had not received GH treatment(n=14) and Group 2, those who showed subnormal growth(n=25). Patients in Group 2 were subdivided into Group 2A, when the patients had not received GH treatment in spite of subnormal growth(n=9) and Group 2B, when GH treatment had been added later on(n=16). There were no differences in the age at diagnosis of GHD, initial height standard deviation score(Ht SDS), body mass index(BMI), peak GH concentration between Group 1 and Group 2. Height velocities in Group 1, 2A, and 2B were 8.1+/-.2 cm/yr, 2.4+/-.2 cm/yr, 2.7+/-.2 cm/yr during the first year of endocrinologic follow-up, 7.1+/-.8 cm/yr, 1.2+/-.1 cm/yr, 7.6+/-.7 cm/yr during the second year, 5.9+/-.0 cm/yr, 2.8+/-.9 cm/yr, 7.3+/-.7 cm/yr during the third year, respectively. BMI changes during the first year of endocrinologic follow-up and postoperative prolactin levels were not significantly different between Group 1 and Group 2A. Postoperative IGF-I and IGFBP-3 levels in Group 1 were significantly higher than those in Group 2A(P<0.05). Both IGFBP-3 and prolactin levels correlated significantly with height velocity in Group 1 and 2A(P=0.004 r=0.64 and P= 0.035 r=0.74 , respectively). CONCLUSION: In this study, growth in children with craniopharyngioma was likely to be associated with IGF-I, IGFBP-3 and prolactin levels. Further studies are needed to unravel other growth promoting factors related to GH independent growth.

Clinical Efficacy of Serum Insulin-like Growth Factor- I (IGF-I), IGF-II, and IGF Binding Protein-3 (IGFBP-3) in Predicting the Prognosis of In Vitro Fertilization and Embryo Transfer / 대한산부인과학회잡지

OBJECTIVE: To evaluate the clinical efficacy of serum insulin-like growth factor-I (IGF-I), IGF-II, and IGF binding protein-3 (IGFBP-3) levels in predicting the prognosis of in vitro fertilization and embryo transfer (IVF-ET). MATERIALS AND METHODS: In 84 patients undergoing IVF-ET, serum levels of IGF-I , IGF-II, and IGFBP-3 were measured using immunoradiometric assay (IRMA) before the gonadotropin administration and on the hCG day of controlled ovarian hyperstimulation (COH). Serum levels of IGFs and IGFBP-3, and the outcomes of IVF-ET were retrospectively analyzed and compared between the pregnant (n=18) and nonpregnant (n=66) groups. RESULTS: There were no significant differences in the outcomes of COH such as total dosage of gonadotropins used, duration of COH, serum estradiol (E2) level on the hCG day, numbers of oocytes retrieved and fertilized, and number of embryos transferred between the pregnant and nonpregnant groups. No differences were found in serum levels of IGF- I , IGF-II, and IGFBP-3, and their ratios before the gonadotropin administration and on the hCG day of COH. Basal serum level of IGF-II was lower with the borderline significance in the pregnant group (796.9+/-159.6 vs. 908.9+/-338.9 ng/ml, p=0.056). The ratio of change in IGF-I to that of IGF-II was significantly higher in the pregnant group (0.066+/-0.489 vs. -0.582+/-2.091, p=0.045). CONCLUSION: Even though basal serum level of IGF-II was lower and the ratio of changes in IGF-I to IGF-II was higher in the pregnant group, serum levels of IGF-I , IGF-II, and IGFBP-3 do not seem to predict the prognosis of IVF-ET. Further investigations are necessary in a larger group of patients to elucidate the clinical efficacy of serum IGFs and IGFBPs levels in predicting the prognosis of IVF-ET.

The Effects of Insulin-like Growth Factor-I (IGF-I) in Mouse Lung Cancer Cells / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Insulin-like growth factor-I (IGF-I) is an important mitogen in many types of malignancies. The purpose of this study was to evaluate the role of the IGF system on cell proliferation and cell death in mouse lung cancer cell lines (3LL). MATERIALS AND METHODS: Northern analysis was performed in 3LL cells. We evaluated the phosphorylation of IGF-I receptor (IGF-IR) with IGF-I stimulation. MTT assay was performed after treating 3LL cells with IGF-I and the treatment effect on cell death in the presence of anticancer drug was investigated. RESULTS: Northern analysis revealed the presence of IGF-I and IGF-IR mRNA expression in 3LL cells. IGF-I increased cellular proliferation in serum free media. IGF-I also stimulated the tyrosine phosphorylation of two proteins: one, with a molecular mass of 95 kDa, was the beta-subunit of IGF-IR; the other, with an approximate molecular mass of 185 kDa, was originally identified as the insulin receptor substrate-I (IRS-I). IGF-I at a low concentration inhibited the cell death induced by adriamycin. CONCLUSION: IGF-I, a mitogen through the phosphorylation of the IGF-IR beta-subunit, acts as a survival factor to inhibit cell death. Therefore, these findings suggest that IGF-I and IGF-IR are involved in both the cell proliferation and cell death associated with cancer cell growth.

The Role of Insulin-like Growth Factor I(IGF-I), and IGF Binding Protein (IGFBP) in Mouse Lung Cancer Cells / 결핵

BACKGROUND: IGF-I is an important mitogen in many types of malignancies. Tumors also express many IGF binding proteins, which modulate IGF action. The purpose of this study was to evaluaste the effect of IGF-I and IGFBP on cell proliferation in mouse lung cancer cells (3LL). METHODS: The cellular proliferation of 3LL with the treatment of growth factors was evaluated using MTT assay. Western ligand blot was performed in order to determine whether 3LL cells secrete IGFBPs and we evaluated the effect of IGFBP on cellular proliferation. RESULTS: The treatment of 3LL cells with IGF-I increased cellular proliferation in a serum free media. Western ligand blot of conditioned medium of 3LL with 125I-IGF-I demonstrated one single major band with an estimated molecular mass of 24 kDa. This band was identified as IGFBP-4 with immunoblot analysis using antisera. The addition of anti-IGFBP-4 antibody to abrogate the effect of IGFBP-4 resulted in increased cellular prolife ration suggesting that IGFBP-4 inhibits cell growth. CONCLUSION: IGF-I increases cellular proliferation, however the secreted IGFBP- 4 has an ingibitory function on cell growth in 3LL. These findings suggest that IGF-I and IGFBP are involved in the cell proliferation.

Malignant solitary fibrous tumor of the pleura causing recurrent hypoglycemia; immunohistochemical stain of insulin-like growth factor i receptor in three cases

We present three cases of malignant solitary fibrous tumors of the pleura (SFTP) that produced recurrent hypoglycemia. Removal of the tumors produced normoglycemia. The tumors were well circumscribed and lobulated, and consisted of firm masses weighing 1,150 g to 1,450 g with the greatest diameter of 15 to 20 cm. The tumors were composed of spindle cells in fascicles or in a haphazard arrangement and were highly cellular and mitotically active (3-8 mitoses/10 high-power fields), showing histologically malignant features. Ultrastructurally, fibroblastic features of the tumor cells were present. Insulin-like growth factors (IGF) have been implicated in the presentation of hypoglycemia. The serum insulin and C-peptide levels were not elevated. Serum IGF-I levels were also low with values of 97.4, 157.1 and 51.9 ng/mL (ref. 125-317 ng/mL), respectively. However, tumor cells were strongly positive for IGF-I receptor on immunohistochemical analysis. It is tempting to speculate that IGF-I contributes to the hypoglycemia, even though the circulating levels were low.

The Diagnostic Utility of Insulin-like Growth Factor Binding Protein-3(IGFBP-3) in Growth Hormone Deficiency and Comparison with Insulin-like Growth Factor-I(IGF-I) / 대한소아내분비학회지

PURPOSE: For the diagnosis of growth hormone(GH) deficiency in short stature, peak growth hormone levels after pharmacologic stimulation are usually used. In this study, we measured serum IGFBP-3, which is a major binding protein in serum and is considered to be GH-IGF-I axis dependent, levels by radioimmuno assay(RIA) in sera from normal short stature(NSS) children, and patients with GH deficiency children to clarify the utility of IGFBP-3 level as a diagnostic marker for GH deficiency, and compare with IGF-I. METHODS: At the department of Pediatrics, Hanyang university hospital from November, 1992 to July, 1995, we selected 31 GH deficiency-suspected children on the base of their growth data and bone age. After GH stimulation with clonidine (100-150microg/m2) and L-dopa(200-250mg/m2), we measured their peak GH levels by the immunoradiometric assay(IRMA) kit(Immunodiagnostic system, UK), IGF-I level by Nichols RIA kit after separated from other plasma constituents with YMC-pack Diol 120 column using high-performance liquid chromatography, and IGFBP-3 level by radioimmuno assay(RIA) kit(Diagnostic system labortories, USA). RESULTS: 1)The mean IGF-I and IGFBP-3 level of eight normal short stature(NSS) in Tanner stage I is 124.3+/-7.3ng/mL, 2,400+/-,500ng/mL respectively. 2)The mean IGF-I and IGFBP-3 level of five partial GH deficient(PGHD) children in Tanner stage I is 163.4+/-8.8ng/mL, 1,800+/-,100ng/mL respectively. 3)The mean IGF-I and IGFBP-3 level of six complete GH deficient(CGHD) children in Tanner stage I is 24.5+/-0.0, 700+/-00ng/mL respectively. There is no significant difference of mean IGF-I and IGFBP-3 levels between NSS and PGHD in Tanner stage I, but the mean IGF-I and IGFBP-3 level is significant difference between NSS and CGHD in Tanner stage I(P<0.05). 4)The sensitivity of IGF-I and IGFBP-3 less than 9 years old for CGHD are 100%. The sensitivity of IGF-I and IGFBP-3 for all age in CGHD is 78, 89% respectively. The sensitivity of IGF-I and IGFBP-3 for GH deficiency in less than 9 years is 60, 80% respectively, and in all age is 55, 65% respectively. The specificity of IGF-I and IGFBP-3 for NSS is 55, 64% respectively. CONCLUSION: Both IGF-I and IGFBP-3 can be useful for screening GH deficiency in Tanner stage I. These two factors are changed according to sexual maturation and nutritional status, but IGFBP-3 has little exogenous effects than IGF-I. Therefore IGFBP-3 may be more sensitive test for diagnosing GH deficiency.

The effect of Insulin like growth factor-I (IGF-I) and IGF binding protein-3 (IGFBP-3) on cellular proliferation in mouse 3T3 fibroblast cells / 결핵

BACKGROUND: Cell growth is a balance between cell proliferation and cell death. Insulin-like growth factor-I(IGF-I), which binds IGF-I receptor (IGF-IR), mediates cellular proliferation as a potent mitogen. IGF binding protein-3 (IGFBP-3) as a circulating major IGFBP can inhibit or enhance the effects of IGF-I on cellular growth by binding IGFs. METHODS: We investigated the expressions of mRNA of IGF-I and IGF-IR by northern blot and phosphorylation of IGF-IR with the treatment of IGF-I by western blot in 3T3 fibroblast cells. The cellular proliferations of 3T3 cells with the treatments of IGF-I were evaluated using 3H-thymidine incorporation and MTT assay. Also to observe the effect of IGFBP-3 on cellular proliferation, 3T3 cells were treated with anti-IGFBP-3 and alpha IR3(monoclonal antibody to IGF-IR) alone or in combination. RESULTS: Our results demonstrated that 3T3 cells showed mRNA expressions of IGF-I and IGF-IR and the IGF-I increased phosphorylation of IGF-IR. The treatments of 3T3 cells with IGF-I increased cellular proliferation in 5% and 1% seruma-containing media, not in serum-free media. The addition of anti-IGFBP-3 to neutralize IGFBP-3 showed 2-fold increase of cellular proliferation, and also co-incubation of anti-IGFBP-3 and alpha IR3 together showed similar increase of cellular proliferation in 3T3 cells. Interestingly, when the cells were prethreated with alpha IR3 for 4 hr, prior to the simultaneous addition of alpha IR3 and anti-IGFBP-3, anti-IGFBP-3-mediated cellular proliferation was decreased to control level. All of these results suggest that free IGF-I released from IGF-I/IGFBP-3 complex would be involved in the cellular proliferation. CONCLUSION: IGF-I action by binding IGF-I.

Expression of Insulin-like Growth Factor I (IGF-I) and Its Binding Proteins in Rat Tissues / 대한소아내분비학회지

covered with liguid nitrozen and pulverized with a pestle. To the powered tissue 5ml of 3.3M formic acid/0.5% Tween 20 was added and centrifuged at 40,000*g for 10 min. An aliquot of supernate was put into C18 sepak minicolumn to eliminates IGF-BPs. Measurement of IGF-I in rat tissues was done by RIA with anti-hIGF-I antibody and hIGF-I(PSIII) standard which was prepared by Drs. L. E. Underwood and J. J. Van Wyk UNC at Chapel Hill, NC, USA and distributed through the National Hormone and Pituitary Distribution Program. Distribution of IGF-I in rat tissue was seen by SDS-PAGE and ligand blotting method. A cDNA library in lambda gt11 of rat liver was used to isolate the cDNA of IGF-I. Phage containing inserts encoding rat IGF-I were identified by hybridization with biotin labeled synthesized oligomer which was the sequence from 1 to 8 aminoacids of known rat IGF-I. The EcoRI inserts were subcloned into PBluescript SK. The nucleotide sequence of both strands was determined by the dideoxy chain termination method. RESULTS: 1)IGF-BPs in tissue extract which could compete with antibody for IGF-I in measureing the IGF-I were eluted at 50Kdalton molecular weight marker using Protein-pak 300SW column. Using C18-sepak minicolumn, IGF-BPs were completely eliminated from tissue extract as much as possible, using Protein-pak 300SW column. 2)The amount of IGF-I in tissues was as folows: liver 575+/-41.6ng/g, lung 552.0+/-40.8ng/g. kidney 503+/-30.8ng/g, heart 449.0+/-30.4ng/g, testis 225+/-18.8ng/g, spleen 146+/-26.4ng/g, muscle 92+/-7.6ng/g and brain 49.0+/-5.8ng/g. The amount of IGF-I in blood was 1403+/-60.8ng/ml. 3)Banding patterns of IGF-BPs in rat tissues extract were obtained using ligand blotting. IGF-BP3 bands at 50 Kdalton molecular weight marker were strongly shown in testis, heart, and lung extracts but not in brain and muscle. IGF-BP1 and 2 band at 30Kdalton molecular weight marker was strongly shown in liver, kidney, spleen, testis, heart and lung. IGF-BP4 band at 21 Kdalton molecular weight marker was weakly shown only in spleen and muscle. 4) The nucleotide sequence of cloned cDNA of rat IGF-I is as follows. 5 10 15 5'----- CC CTT TGC GGG GCT GAG CTG GTG GAC GCT CTT CAG TTC GTG TGT 20 25 30 -GGA CCA AGG GGC TTT TAC TTC AAC AAG CCC ACA GGC TAT GGC- 35 40 45 -TCC AGC ATT CGG AGG GCA CCA CAG ACG GGC ATT GTG GAT GAG------3 CONCLUSION: This study suggests that tissue extraction method for IGF-I from tissues and elimination of IGF-BPs using C18 sepak minicolumn is suitable for measuring in large numbers of samples. Expression of IGF-I and IGF-BPs in multiple tissues suggests some phsiologic function at each tissue level. Subcloning of cDNA of exon 3 and 4 of IGF-I was useful for studying regulation of IGF-IA and IB mRNA in rat tissue.

The Growth Promoting Effect of Insulin-like Growth Factor-I(IGF-I) Purified from Human Serum F208 on the Rat Rib Chondrocytes / 대한소아내분비학회지

PURPOSE:The pathogenesis of short stature in growth hormone(GH) deficiency is believed to be based on the growth failure of growth plate chondrocytes by reduced growth hormone dependent insulin-like growth factor- I (IGF- I ) level in serum. Therefore, author studied the growth promoting effect of IGF- I purified from human serum on the chondrocytes, cultured from rat rib cartilage. METHODS:Rat rib cartilage were treated with type II collagenase and hyaluronidase and were cultured in Ham's F-12 culture media containing 10% fetal calf serum. Growth promoting effect of IGF- I was measured by MTT dye by adding 20ng/ml IGF- I purified by protein-diol 120 column(YMC Co, Japan) from human serum, to 1*104 cultured chondrocytes separated into each of 96 well culture vessel. RESULTS: 1) When elution time of biotin labeled IGF- I by protein pak 300sw column was compared to elution time of standard molecular weight, IGF- I exists as large complex of 150Kd and small complex of 50Kd with free 7Kd form in serum before acid treatment. After acid treatment, IGF- I exists as small complex of 50Kd with free 7Kd form. 2) IGF- I purified from blood samples, as compared to genetic engineering product standard IGF- I , showed good parallelism in competition inhibition curve by purity analysis utilizing IGF- I antibody, and thus it is assumed that complex protein as inhibiting factor for purified IGF- I does not exist. Furthermore, complex protein was not present on the Western ligand method using biotin-labeled IGF- I ligand after purified IGF- I transferred to nitrocellulose paper following SDS-PAGE electrophoresis. 3)IGF- I of 20ng/ml showed 30% growth promoting effect, when rat rib chondrocyte culture with Dulbeco's modified Eagles medium(DMEM) is considered to show maximum growth promoting effect, while with pure culture medium, DMEM, showing minimum effect. CONCLUSIONS:The results of this study suggest that IGF- I purified by this method assumes the role of growth promoting effect on the chondrocytes, and that the described method of radioimmuno assay of IGF- I also could effectively remove inhibiting protein complex, therefore allowing more accurate assay.