BACKGROUND: Sex steroid hormones are believed to play an important role in the genesis and growth of uterine myoma. Several studies suggest a possible role of insulin-like growth factor(IGF) as a mediator of estradiol in uterine myama. We have recently demonstrated that some IGF binding proteins(IGFRPs) messenger ribonucleic acid(mRNA) expressions in myoma are dependent on the in vivo esttogen status. The purposes of this study are to evaluate the in vitro effects of sex steroid hormones including estrogen on the IGFBPs gene expression in tissues from uterine myoma and adjacent normal myometrium. METHODS: Tissues from myoma and adjacent normal myometrium of patients with uterine myoma during early proliferative phase of menstrual cycle were cultured in the absence(control) and presence of 17b-estradiol(10M/L) or/and progesterone(10M/L) for 3 days. The IGFBPs mRNA expressions in these explants were analyzed by Nothern blot using specific human complementary deoxyribonucleic acid(cDNA) probes. RESULTS: The addition of 17b-estradiol, progesterone alone and in combination to conditioned media of explants from myoma and adjacent normal myornetrium did not result in any changes in the expression of IGFBP-2, IGFBP-4, IGFBP-5, and IGFBP-6 mRNA. With progesterone addtion, lGFBP-3 rnRNA expression was significantly reduced in myoma explant but not in adjacent ncemal myometrium explant. There was no significant change in the IGFBP-3 mRNA expression with 17b-estradiol and with the combination of both 17b-estradiol and progesterone. CONCLUSION: 17b-estradiol does not affect IGFBPs gene expression in the myoma and adjacent normal myometrium explant regardless of the presence of progesterone in vitro. However progesterone alone induces a decrease in IGFBP-3 synthesis in myoma explant.
Animals ; Culture Media, Conditioned ; Estradiol ; Estrogens ; Female ; Gene Expression ; Gonadal Steroid Hormones ; Humans* ; Insulin-Like Growth Factor Binding Protein 2 ; Insulin-Like Growth Factor Binding Protein 3 ; Insulin-Like Growth Factor Binding Protein 4 ; Insulin-Like Growth Factor Binding Protein 5 ; Insulin-Like Growth Factor Binding Protein 6 ; Insulin-Like Growth Factor Binding Proteins ; Leiomyoma* ; Menstrual Cycle ; Mice ; Myoma ; Myometrium* ; Progesterone ; RNA, Messenger*

BACKGROUND: Uterine leiomyoma is the most common pelvic tumor, occurring in 20-25% of women in reproductive age. Gonadotropin releasing hormone agonist (GnRHa) has been reognized as a temporary medical management for this disorder. The etiology of these tumors is unknown but it has been shown that the insulin-like growth factors (IGF-I, IGF-II) are promoters of growth in nongynecologic tumors. Several recent studies have suggested the possible role of IGFs in human leiomyoma growth. The IGF binding proteins (IGFBPs) are believed to modulate actions of IGF and to have IGF-independent actions. The purpose of this study was to evaluate the type of IGF and IGFBP which may be involved in leiomyoma growth and to investigate a possible IGF related mechanism of action of GnRHa. METHOD: The IGFs and IGFBPs were measured by double antibody radioimmunoassay, western ligand blot and immunoprecipitation in the tissue cytosols of normal uterine myometria (n=15), nontumorous myometria adjacent to a leiomyoma and leiomyoma from patients nontreated (n=15) and treated (n=10) with GnRHa. RESULTS: The mean IGF-I and IGF-II level were significantly higher in leiomyoma from untreated patients than in the adjacent myometrium and normal myometrium but no significant differences in these IGF levels between normal myometrium and adjacent myometrium were noted. The IGFBP-2, IGFBP-3 and 26kDa IGFBP were detected variably but IGFBP-4 was consistently present in all tissues. There were no significant differences in the relative intensity for IGFBP-4 and the frequency of IGFBPs between leiomyoma, adjacent myometrium and normal myometrium from untreated patients. The IGF-I, IGF-II levels and the relative intensity of IGFBP-4 in leiomyoma from GnRHa-treated patients were significantly lower than those in untreated patients, but these levels in the adjacent myometrium were comparable. The frequency of each IGFBP in leiomyoma and the adjacent myornetrium from GnRHa-treated patients did not significantly differ from untreated patients. CONCLUSION: Both IGF-I and IGF-II are involved in the growth of leiomyoma and GnRHa may in part act to decrease size of leiomyoma by regulating the local levels of IGF-I, IGF-II and IGFBP-4.
Animals ; Cytosol ; Female ; Gonadotropin-Releasing Hormone ; Humans ; Immunoprecipitation ; Insulin-Like Growth Factor Binding Protein 2 ; Insulin-Like Growth Factor Binding Protein 3 ; Insulin-Like Growth Factor Binding Protein 4 ; Insulin-Like Growth Factor Binding Proteins ; Insulin-Like Growth Factor I ; Insulin-Like Growth Factor II ; Leiomyoma* ; Mice ; Myometrium ; Radioimmunoassay ; Somatomedins*

PURPOSE: PTEN/MMAC1, a novel tumor suppressor gene, is mutated in a variety of advanced and metastatic cancers. It acts as a phosphatase, and thereby, regulates the PI-3 kinase/Akt pathway. In this study, we examined to evaluate the new function of anti-tumor effects of PTEN/MMAC1 through the regulation of the IGFs-IGFBPs in gastric cancer cells. METHODS: PTEN/MMAC1 was expressed in an adenovirus-mediated gene delivery system and introduced into gastric cancer cells(SNU-484 & SNU-668) in vitro. The effect of cell growth and the expression of IGFs and IGFBPs after Ad/PTEN infection was analyzed by MTT assay, RT-PCR and Western immunoblot. RESULTS: Ad/PTEN infected cells were inhibited in cell growth compared with moak cells and Ad/ LacZ infected cells. Overexpression of PTEN/MMAC1 induced decrease in expression of IGF-I, -II and IGF-I receptors which are known as growth prompt molecules in a variety of cancers. Of the six IGFBPs, the expressions of IGFBP-4 and IGFBP-6 were decreased in Ad/PTEN infected cells. In contrast, IGFBP-3 expression was markedly increased by up to 3-fold in Ad/PTEN infected cells. Overexpression of PTEN/MMAC1 inhibited the activation of Akt/PKB pathway, but had no effect on the MAPK pathway. CONCLUSION: These findings suggest that the tumor suppressor function of PTEN/MMAC1 is, at least in part, mediated through the down-regulation of IGF-I abd IGF-II, and up-regulation of IGFBP-3 in gastric cancer cells by the inhibition of PI-3 kinase pathway.
Blotting, Western ; Down-Regulation ; Gene Transfer Techniques ; Genes, Tumor Suppressor ; Insulin-Like Growth Factor Binding Protein 3 ; Insulin-Like Growth Factor Binding Protein 4 ; Insulin-Like Growth Factor Binding Protein 6 ; Insulin-Like Growth Factor Binding Proteins* ; Insulin-Like Growth Factor I ; Insulin-Like Growth Factor II ; Phosphatidylinositol 3-Kinase ; Phosphatidylinositol 3-Kinases ; Receptor, IGF Type 1 ; Stomach Neoplasms ; Up-Regulation

The insulin-like growth factor(IGF) system consists of IGFs, their receptor and binding proteins(IGFBPs). The IGFs are important growth factors in the regulation of fetal growth. Since IGFBPs control IGF actions, the IGFBPs themselves may also be important in fetal growth and development. The goals of this study are to investigate the profiles of IGFBPs in cord sera of appropriate-for-gestatinal age(AGA, n=27), small-for-gestatinal age(SGA, n=14), large-for-gestatinal age(LGA, n=10) infants and preterm(PT, n=14) infants and to evaluate the relationship between these IGFBP levels and gestational weeks and birth weight and between total IGFBP levels in cord sera and paired maternal sera(n=65). The IGFBPs were analyzed by Western ligand blot and immunoprecipitation. In cord sera of AGA infants IGFBPs with molecular weight with 37/43 kilocatons(kDa; IGFBP-3), 31 kDa(IGFBP-2), 26 kDa(IGFBP-1), 24 kDa(IGFBP-4) were detected. In cord sera of LGA infants there was a significant increase in IGFBP-3 levels and a reduction of IGFBP-1 and IGFBP-4 levels compared with those in AGA infants. SGA infants had significantly higher IGFBP-1 and IGFBP-2 levels in cord sera than AGA infants. There was a similiar trend in IGFBP-1 levels in cord sera of PT infants. The relative proportion of IGFBP-4 in cord sera of SGA and PT infants was significantly higher than that of AGA infants. There was no significant correlation beween total IGFBP levels in cord sera and paired maternal sera. The ratios of total IGFBP in cord sera to that in maternal sera to that in maternal sera were significantly higher in SGA and PT infants than in AGA infants. The IGFBP-1 and IGFBP-2 levels correlated with birth weight but did not correlate with gestational weeks. These data suggest that there is an unique profile of IGFBPs in cord sera of infants according to their weight, and that IGFBPs may play a major role in the control of fetal growth.
Birth Weight ; Fetal Development ; Gestational Age* ; Humans ; Immunoprecipitation ; Infant* ; Infant, Newborn ; Infant, Premature* ; Insulin-Like Growth Factor Binding Protein 1 ; Insulin-Like Growth Factor Binding Protein 2 ; Insulin-Like Growth Factor Binding Protein 3 ; Insulin-Like Growth Factor Binding Protein 4 ; Insulin-Like Growth Factor Binding Proteins ; Intercellular Signaling Peptides and Proteins ; Molecular Weight

PURPOSE: Insulin-like growth factor (IGF)-I and II are potent mitogens, postulated to exert autocrine and paracrine effects on growth regulation in human gastric cancer. In this study, we evaluated the expression of IGF-I, -II and IGFBPs in a panel of human gastric cancer cell lines. We also evaluated whether high expression of IGFBP-3 in human gastric cancer cells may increase the sensitivity to the anti-proliferative agents. MATERIALS AND METHODS: 10 human korean gastric cancer ceIl lines and 1 Caucacian gastric adenocarcinoma cell line were used for this study. IGF and IGFBP expressions were evaluated by RT-PCR. IGFBP proteins in conditioned media were detected by Western Ligand Blot. Cell survival after treatment of anti-proliferative agents was assessed by MTT assay. RESULTS: IGF-I and II were expressed in all gastric cancer cell lines. In addition, IGF-I and II stimulated the proliferation of gastric cancer cells. The expression of IGFBP-2 was found in all gastric cancer cell lines. IGFBP-4 was expressed in the most of cell lines. IGFBP-3, -4 and -6 were expressed in about 50% of cell lines. The growth inhibition of IGFBP-3 expressing cells by anti- proliferative agents was more significant than that of IGFBP-3 nonexpressing cells. Cell growth inhibition with treatment of these agents was accompanied by increased IGFBP-3 mRNA level. CONCLUSION: These data confirm that IGF-I, -II, and certain IGFBPs were expressed in gastric cancer cells, and gastric cancer cells show the differential growth inhibition by anti-proliferative agents. The differential growth inhibitory effect of anti-proliferative agents is, at least in part, mediated through up-regulation of IGFBP-3 expression.
Adenocarcinoma ; Carrier Proteins* ; Cell Line* ; Cell Survival ; Culture Media, Conditioned ; Humans ; Insulin-Like Growth Factor Binding Protein 2 ; Insulin-Like Growth Factor Binding Protein 3* ; Insulin-Like Growth Factor Binding Protein 4 ; Insulin-Like Growth Factor Binding Proteins ; Insulin-Like Growth Factor I ; Mitogens ; RNA, Messenger ; Stomach Neoplasms* ; Up-Regulation

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.
Animals ; Carrier Proteins* ; Cell Proliferation ; Culture Media, Conditioned ; Culture Media, Serum-Free ; Immune Sera ; Insulin-Like Growth Factor Binding Protein 4 ; Insulin-Like Growth Factor Binding Proteins ; Insulin-Like Growth Factor I ; Intercellular Signaling Peptides and Proteins ; Lung Neoplasms* ; Lung* ; Mice*

OBJECTIVES: To evaluate changes in serum insulin-like growth factor binding protein(IGFBP)s profiles during the controlled hyperstimulation cycle and to compare IGFBPs profiles and their protease activity in follicular fluid(FF) from polycystic ovary and preovulatory follicle containing mature oocyte. METHODS: IGFBPs and their protease activity were measured by western ligand blot, immunoprecipitation and mixing test in fasting blood samples obtained throughout ovarian hyperstimulation cycle from 33 patients undergoing in vitro fertilization (IVF)-embryo transfer, and in FF from polycystic ovary(n=10) and IVF follicle(n=30) containing mature oocyte. Serum 17 -estradiol was also determined by radioimmunoassay. RESULTS: Type and molecular weight of serum IGFBP did not changed during the controlled hyperstimulation cycle. No significant variations in the relative proportion, level of IGFBPs and their protease activity were found throughout the stimulated cycle. IGFBP-4 levels in FF from polycystic ovary were significantly higher than in FF from IVF follicle and IGFBP -2 levels also showed a similar trend, but no significant differences in other IGFBP profiles and their protease activity were noted. IGFBP-4 protease activity was significantly higher in the latter follicle than in the former follicle. There were significant correlations between serum IGFBP-1, IGFBP-3 and 17 -estradiol levels. Correlation between serum and FF IGFBP except IGFBP-1 was not significant. CONCLUSIONS: IGFBPs have regulatory functions in ovary through an paracrine and autocrine rather than endocrine mechanism during the controlled hyperstimulation cycle. IGFBP-4 and its protease activity in FF may be involved in the follicle growth.
Fasting ; Female ; Fertilization in Vitro ; Follicular Fluid* ; Humans ; Immunoprecipitation ; Insulin-Like Growth Factor Binding Protein 1 ; Insulin-Like Growth Factor Binding Protein 3 ; Insulin-Like Growth Factor Binding Protein 4 ; Insulin-Like Growth Factor Binding Proteins ; Molecular Weight ; Oocytes ; Ovary ; Pregnancy-Associated Plasma Protein-A ; Radioimmunoassay

Cows with different Insulin-like Growth Factor-I (IGF-I) concentrations showed comparable expression levels of hepatic growth hormone receptor (GHR). Suppressor of cytokine signaling 2 (SOCS2), could be responsible for additional inhibition of the GHR signal cascade. The aims were to monitor cows with high or low antepartal IGF-I concentrations (IGF-I(high) or IGF-I(low)), evaluate the interrelationships of endocrine endpoints, and measure hepatic SOCS2 expression. Dairy cows (n = 20) were selected (240 to 254 days after artificial insemination (AI)). Blood samples were drawn daily (day -17 until calving) and IGF-I, GH, insulin, thyroid hormones, estradiol, and progesterone concentrations were measured. Liver biopsies were taken (day 264 +/- 1 after AI and postpartum) to measure mRNA expression (IGF-I, IGFBP-2, IGFBP-3, IGFBP-4, acid labile subunit (ALS), SOCS2, deiodinase1, GHR1A). IGF-I concentrations in the two groups were different (p < 0.0001). However, GH concentrations and GHR1A mRNA expression were comparable (p > 0.05). Thyroxine levels and ALS expression were higher in the IGF-I(high) cows compared to IGF-I(low) cows. Estradiol concentration tended to be greater in the IGF-I(low) group (p = 0.06). It was hypothesized that low IGF-I levels are associated with enhanced SOCS2 expression although this could not be decisively confirmed by the present study.
Animals ; Cattle ; Estradiol/blood ; Female ; Growth Hormone/blood ; Insulin/blood ; Insulin-Like Growth Factor Binding Protein 2/analysis ; Insulin-Like Growth Factor Binding Protein 3/analysis ; Insulin-Like Growth Factor Binding Protein 4/analysis ; Insulin-Like Growth Factor I/*analysis/physiology ; Liver/chemistry ; Pregnancy/metabolism/physiology ; Pregnancy, Animal/*metabolism/physiology ; Progesterone/blood ; Suppressor of Cytokine Signaling Proteins/analysis ; Thyroid Hormones/blood

Gonadotropin releasing hormone agonist(GnRHa) has been used as a temporaruy medical management for uterine myoma. Insulin-like growth factors(IGFs) has been supposed to play an etiological role in myoma growth and their action is modulated by IGF-binding proteins(IGFBPs). The purpose of this study is to evaluate the effects of GnRHGa on the IGFBPs production in the explant culture of adjacent myometrium and uterine myoma. The adjacent myometrium and myoma explants from patients treated(n=10) and nontreated(n=16) with GnRHa were cultured under serum free condition.Some explants from untreated patients were also cultured in the presence of GnRHa(10(-7) M/L). Western ligand blot and immunoprecipitation showed the presence of 276 kilodalton IGFBP, IGFBP-2, IGFBP-3 and IGFBP-4 in culture media of these explants and IGFBP-4 only was consistently present in culture media of all explants. There were no significant differences in the frequency of each IGFBP between adjacent myometrium and myoma explant cultures regardless of the type of GnRHa exposure. The relative IGFBP-4 level in explant cultures of myoma from patients treated with GnRHa was significantly higher than that from untreated patients.Addition of GnrHa in the media of myoma explants did not result in an significant change in the relative IGFBP-4 level compared with in vitro untreated explant cultures. Our data suggest that Gnrha may act through indirect involvement in the EGFBP-4 production in myoma tissue.
Animals ; Culture Media ; Female ; Gonadotropin-Releasing Hormone* ; Gonadotropins* ; Humans* ; Immunoprecipitation ; Insulin-Like Growth Factor Binding Protein 2 ; Insulin-Like Growth Factor Binding Protein 3 ; Insulin-Like Growth Factor Binding Protein 4 ; Insulin-Like Growth Factor Binding Proteins ; Leiomyoma* ; Mice ; Myoma ; Myometrium*

Gonadotropin releasing hormone agonist(GnRHa) has been used as a temporaruy medical management for uterine myoma. Insulin-like growth factors(IGFs) has been supposed to play an etiological role in myoma growth and their action is modulated by IGF-binding proteins(IGFBPs). The purpose of this study is to evaluate the effects of GnRHGa on the IGFBPs production in the explant culture of adjacent myometrium and uterine myoma. The adjacent myometrium and myoma explants from patients treated(n=10) and nontreated(n=16) with GnRHa were cultured under serum free condition.Some explants from untreated patients were also cultured in the presence of GnRHa(10(-7) M/L). Western ligand blot and immunoprecipitation showed the presence of 276 kilodalton IGFBP, IGFBP-2, IGFBP-3 and IGFBP-4 in culture media of these explants and IGFBP-4 only was consistently present in culture media of all explants. There were no significant differences in the frequency of each IGFBP between adjacent myometrium and myoma explant cultures regardless of the type of GnRHa exposure. The relative IGFBP-4 level in explant cultures of myoma from patients treated with GnRHa was significantly higher than that from untreated patients.Addition of GnrHa in the media of myoma explants did not result in an significant change in the relative IGFBP-4 level compared with in vitro untreated explant cultures. Our data suggest that Gnrha may act through indirect involvement in the EGFBP-4 production in myoma tissue.
Animals ; Culture Media ; Female ; Gonadotropin-Releasing Hormone* ; Gonadotropins* ; Humans* ; Immunoprecipitation ; Insulin-Like Growth Factor Binding Protein 2 ; Insulin-Like Growth Factor Binding Protein 3 ; Insulin-Like Growth Factor Binding Protein 4 ; Insulin-Like Growth Factor Binding Proteins ; Leiomyoma* ; Mice ; Myoma ; Myometrium*