Gallbladder cancer-associated fibroblasts promote the migration of lymphatic endothelial cells via releasing IGFBP3 / 西安交通大学学报(医学版)

【Objective】 To investigate the effects of gallbladder cancer-associated fibroblasts (CAFs) on the migration of lymphatic endothelial cells (LECs) so as to elucidate the molecular mechanisms involved. 【Methods】 The CAFs and normal fibroblasts (NFs) were extracted by enzymatic digestion, and the supernatant (CM) of CAFs and NFs was collected. The levels of IL-6, IGFBP3 and other related cytokines were detected by semi-quantitative protein factor microarray and ELISA. The expressions of α-SMA (CAFs maker) and IGFBP3 in gallbladder cancer and para-cancer tissues were detected by immunohistochemistry, and the correlation of α-SMA and IGFBP3 expressions with clinicopathological characteristics were analyzed. LECs were cultured and divided into serum-free medium group (control group), CAF-CM co-culture group, NF-CM co-culture group, IGFBP3 group, and CAF-CM+IGFBP3 inhibitor (2-Deoxy-D-glucose, 2-DG) group according to different treatment. Transwell migration assays and wound healing assays were applied to analyze the migration ability of LECs under different treatment. The expressions of E-cadherin, N-cadherin and Vimentin were detected by Western blotting. 【Results】 Protein factor microarray and ELISA showed that the concentration of IGFBP3 in CAF-CM was significantly increased, and the expression of α-SMA was significantly related to lymph node metastasis, advanced TNM stage and expression of IGFBP3. IGFBP3 secreted from CAF-CM significantly promoted LECs migration, up-regulated the expression of N-cadherin and Vimentin, and down-regulated the expression of E-cadherin. Treatment with IGFBP3 inhibitor 2-DG could reverse the effect of CAF-CM on migration of LECs and related protein expressions. 【Conclusion】 Gallbladder CAFs promote the migration of LECs via releasing IGFBP3, which affects EMT transformation.

The expression and significance of IGFBP-3 in salivary gland pleomorphic adenoma / 实用口腔医学杂志

Objective:To investigate the expression of insulin-like growth factor binding protein 3 (IGFBP-3) in salivary pleomorphic adenoma(SPA).Methods:The expression of IGFBP-3 protein in 40 cases of SPA(group SPA),40 of normal glandular tissue(group N) and 10 of salivary gland malignant tumor(group CA) was detected by Western blot.The expression of IGFBP-3 mRNA in 50 cases of SPA,50 of salivary gland normal tissue and 10 of CA was detected by qRT-PCR.Results:The expression(A value) of IGFBP-3 protein in group N,SPA and CA was 8.54 ± 3.95,4.78 ± 2.07,3.63 ± 2.27 respectively.The expression ration of IGFBP-3 mRNA of group N vs SPA or CA,P ＜ 0.05;SPA vs CA,P ＞ 0.05 (SPA/N was 0.654 ± 0.387,CA/N:0.452 ± 0.229) respectively,but showed no significance difference between SPA and the CA groups(P ＞ 0.05).Difference of IGFBP-3 mRNA expression was observed with different envelope infiltration of SPA (P ＜ 0.05),no significant difference was observed in different age,gender or relapse groups.Conclusion:IGFBP-3 Low expression of IGFBP-3 in pleomorphic adenomas may reduce the antagonism of IGF-1R,causing the proliferation of tumor cells and promote tumor formation.

Promoter -202 A/C Polymorphism of Insulin-like Growth Factor Binding Protein-3 Gene and Non-small Cell Lung Cancer Risk / 결핵및호흡기질환

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 inter- individual 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 poly?morphic 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 freque?ncies 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 (pAC>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.

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.

EFFECTS OF IGFBP-3/RXR? OR STAT-1 SIGNALING PATHWAY ON A?_(1-42) INDUCED APOPTOSIS IN RAT HIPPOCAMPUS NEURONS / 解剖学报

Objective To investigate the effects of IGFBP-3,RXR? and STAT-1 on A?_ 1-42 induced apoptosis in rat hippocampus neurons.Methods Apoptosis was induced by fibrillar A?_ 1-42.The percentage of neurons apoptosis was evaluated by microscopy after staining with TUNEL/DAPI.IGFBP-3 and RXR? positive neurons were observed by immunofluorescence.The expression of RXR? and STAT-1 protein were detected by Western blotting.Results After treatment with 20?mol/L A?_ 1-42 for 24 hours,the apoptotic hippocampus neurons were shown by TUNEL/DAPI assay.The percentage of apoptotic neurons was increased in a time-dependent manner.During the development of apoptosis,both the percentage of IGFBP-3/RXR? positive neurons and the expression of RXR? protein increased markedly after 3-6hours(P

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 Characteristics of Insulin-like Growth Factor Binding Protein-3(IGFBP-3) in Serum; The Diagnostic Utility of IGFBP-3 in Growth Hormone Deficiency

PURPOSE: For 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. METHODS: At the department of Pediatrics, Hanyang University Hospital from November, 1992 to July, 1995, we selected 32 GH deficiency-suspected children on the base of their growth data and bone age. After GH stimulation with clonidine(100-150mug/m2) and L-dopa(200-250 mg/m2), we measured their peak GH levels by the immunoradiometric assay(IRMA) kit(Immunodiagnostic system, UK), IGFBP-3 level by radioimmuno assay(RIA) kit(Diagnostic system labortories, USA). RESULTS: 1) The mean IGFBP-3 levels of eight normal short stature(NSS) in Tanner stage I is 2.4+/-1.5mug/ml and their stimulated mean peak GH level is 18.7+/-7.5ng/ml. However, one child in Tanner stage I with nutritional deficiency, IGFBP-3 level is 0.717mug/ml and stimulated peak GH level is 12.2ng/ml. And the mean IGFBP-3 and peak GH levels of two Tanner stage II NSS are 2.2+/-1.2mug/ml and 14.3+/-5.2ng/ml, respectively. 2) The mean IGFBP-3 level of five partial GH deficient(PGHD) children in Tanner stage I is 1.8+/-1.1mug/ml, and their stimulated mean peak GH level is 8.2+/-1.3ng/ml. The mean IGFBP-3 level of five PGHDs in Tanner stage II is 2.2+/-0.8mug/ml, and their stimulated mean peak GH level is 7.5+/-1.5ng/ml. 3) The mean IGFBP-3 level of six complete GH deficient(CGHD) children in Tanner stage I is 0.7+/-0.6mug/ml, and their stimulated peak GH level is 1.0+/-1.2ng/ml. The mean IGFBP-3 level of three complete GH deficient(CGHD) children in Tanner stage II is 2.2+/-0.2mug/ml, and their stimulated peak GH level is 2.5+/-1.4ng/ml. Only one CGHD child in Tanner stageIII, IGFBP-3 level is 5.943mug/ml, and his stimulated peak GH level is 3.3ng/ml. 4) There is no significant difference of mean IGFBP-3 levels between NSS and PGHD in Tanner stage I, but the mean IGFBP-3 level is significant difference between NSS and CGHD in Tanner stage I(p<0.05). 5) The sensitivity of IGFBP-3 for CGHD and PGHD less than 9 years old is 83%, 75% and for all age is 80%, 55%, respectively. The sensitivity of IGFBP-3 for GH deficiency in less than and older than 9 years is 80%, 67%, respectively. The specificity of IGFBP-3 for NSS is 64%. CONCLUSIONS: Because serum IGFBP-3 levels may increased during puberty due to mechanisms independent of the GH-IGF-I axis, it is difficult to distinguish GH deficiency from NSS in older children, but CGHD in Tanner stage I, we may use the basal plasma IGFBP-3 level as a screening test for diagnosing GH deficiency.