Insulin-like growth factor-binding protein-3 inhibits IGF-1-induced proliferation of human hepatocellular carcinoma cells

OBJECTIVE Basic fibroblast growth factor(b FGF)and platelet-derived growth factor(PDGF)produced by hepatocellular carcinoma(HCC)cells are responsible for the cell growth.Accumulating evidence shows that insulin-like growth factor-binding protein-3(IGFBP-3)suppresses HCC cell proliferation in both IGF-dependent and independent manners.The present study is to investigate whether treatment with exogenous IGFBP-3 inhibits bF GF and PDGF production and the cell proliferation of HCC cells.METHODS Cell Counting Kit 8 assay were designed to detect HCC cell proliferation,transcription factor early growth response-1(EGR1)involving in IGFBP-3 regulation of b FGF and PDGF were detected by RT-PCR and Western blot assays.Western blot assay was adopted to detect the IGFBP-3 regulating insulin-like growth factor 1 receptor(IGF-1R)signaling pathway.RESULTS The present study demonstrates that IGFBP-3 suppressed IGF-1-induced b FGF and PDGF expression while it does not affect their expression in the absence of IGF-1.To delineate the underlying mechanism,Western-blot and RT-PCR assays confirmed that the transcription factor early growth response protein 1(EGR1)is involved in IGFBP-3 regulation of b FGF and PDGF.IGFBP-3 inhibition of type 1 insulin-like growth factor receptor(IGF1R),ERK and AKT activation is IGF-1-dependent.Furthermore,transient transfection with constitutively activated AKT or MEK partially blocks the IGFBP-3 inhibition of EGR1,b FGF and PDGF expression.CONCLUSION In conclusion,these findings suggest that IGFBP-3suppresses transcription of EGR1 and its target genes b FGF and PDGF through inhibiting IGF-1-dependent ERK and AKT activation.It demonstrates the importance of IGFBP-3 in the regulation of HCC cell proliferation,suggesting that IGFBP-3 could be a target for the treatment of HCC.

Significance of highly phosphorylated insulin-like growth factor binding protein-1 and cervical length for prediction of preterm delivery in twin pregnancies

BACKGROUND A twin pregnancy can carry greater risks than singleton pregnancies.About 60 in 100 twin pregnancies result in spontaneous birth before 37 wk,which is associated with several complications in the premature babies.Clinical detection of biomarkers may help to predict the possibility of premature birth so that corresponding interventions can be given to the pregnant women in a timely manner,in order to reduce the risk of preterm birth and improve the outcomes of the newborn infants.AIM To explore the clinical value of transvaginal ultrasound measurement of cervical length combined with insulin-like growth factor binding protein-1(IGFBP-1)hyperphosphorylation in cervical secretions as predictors of preterm delivery in twin pregnancies.METHODS A total of 254 pregnant women with twin pregnancies,who were admitted to Hainan General Hospital and underwent maternity examination,were selected as the study subjects from January 2015 to December 2018.All participants received transvaginal ultrasound measurement of cervical length and phosphorylated IGFBP-1(phIGFBP-1)test between 24 and 34 wk gestation.The pregnancy outcomes were analyzed.RESULTS Of the women with a positive phIGFBP-1 test result,preterm birth rate was higher in those with a cervical length≤25 mm than those with a cervical length>25 mm(all P<0.05).Similarly,in women with a negative phIGFBP-1 test result,preterm birth rate was higher in those with a cervical length≤25 mm than those with a cervical length>25 mm(all P<0.05).The sensitivity,specificity,and positive and negative predictive values of the phIGFBP-1 test combined with the cervical length test were 95.71%,91.21%,95.12%and 92.22%,respectively,for the prediction of preterm birth.CONCLUSION Cervical length combined with phIGFBP-1 tests is of value for the prediction of outcomes of preterm delivery in twin pregnancies.

Cloning and Sequence Analysis of IGFBP-1 Gene in Sheep

In this study, using Liangshan semi-fine wool sheep as an experimental material, full-length CDS sequence of IGFBP-1 gene was cloned with RT-PCR method and analyzed with bioinformatics methods. The results showed that the full-length CDS sequence of IGFBP-1 gene in Liangshan semi-fine wool sheep was 792 bp in length, encoding 263 amino acids. The CDS sequence shared 97%, 76% and 74% homology with bovine, human and rat, respectively; the amino acid sequence shared 97% , 69% and 71%, respectively. The GenBank accession number was FJ589639.1. The amino acid molecular weight of IGFBP-1 was 27.8 kD, and the theoretical isoelectric point （pl） was 5.99. The result of phylogenetic analysis showed that IGFBP-1 gene in Liangshan semi-fine wool sheep exhibited close phylogenetic relationships with bovine, goat and other mammals, and distant phylogenetic relationships with chicken and fish. IGFBP-1 gene had obvious hy- drophobic and hydrophilic regions, harboring one signal peptide, one transmembrane region, 16 phosphorylation sites, six N-glycesylation sites and eight O-glycosy- lation sites. The result of secondary structure analysis showed that the random coil, a-helix and 13-sheet regions accounted for 61.98%, 24.33% and 13.69%, re- spectively. The result of tertiary structure analysis showed that IGFBP-1 harbors an IGFBP_N domain and a thyrnglobulin type-1 domain. This study laid a solid foundation for further investigating the function of IGFBP-1 gene in sheep.

Possible roles of insulin, IGF-1 and IGFBPs in initiation and progression of colorectal cancer

AIM: To investigate the roles of serum insulin, insulin-like growth factor-1 (IGF-1), and insulin-like growth factor binding proteins (IGFBPs) in the initiation and progression of colorectal cancer.

Combination of insulin-like growth factor-1, IGF binding protein-3, chromogranin A and prostate specific antigen can improve the detection of prostate cancer

Aim:Prostate cancer(PCa)is the second most prevalent male cancer worldwide and designated the sixth most frequent male cancer in Arab countries.Although prostate specific antigen(PSA)has become the best and most valuable biomarker for screening of PCa,elevated levels of PSA can reflect the presence of malignant cells but can overlap with benign prostatic diseases.There is a necessity to develop and improve current tools for early detection and diagnosis of PCa.This study was done to evaluate the validation of serum insulin-like growth factor-1(IGF-1),IGF binding protein-3(IGFBP-3),chromogranin A(CgA)and combination with PSA in treatment of benign prostatic hyperplasia(BPH)and PCa patients.Methods:The study included 72 patients with PCa,70 BPH patients and 56 healthy male subjects of matched age.Full history and clinical data were recorded for all subjects.Results:Serum PSA attained sensitivity of 84%at 82%specificity with an accuracy of 83%,although IGF-1,IGFBP-3 and CgA did not recognize PCa patients.Conclusion:Combinations of IGF-1 and IGFBP-3 biomarkers with PSA were effectively differentiated between PCa and control groups as well as improving the overall value of sensitivity,specificity and diagnostic accuracy of PCa to 85%and 86%for IGF-1/PSA and IGFP-3/PSA respectively.

Cloning and Sequence Analysis of IGFBP-7 Gene in Sheep

In this study, full-length CDS sequence of IGFBP-7 gene was cloned from Liangshan semi-fine wool sheep with RT-PCR method and analyzed with bioinformatics methods. The results showed that the full-length CDS sequence of IGFBP-7 gene in Liangshan semi-fine wool sheep was 846 bp in length, encoding 282 amino acids. The CDS sequence shared 99%, 95% and 90% homology with bovine, human and rat, respectively; the amino acid sequence shared 98%, 93% and 89%, respectively. The GenBank accession number was FJ589640.1. The amino acid molecular weight of IGFBP-7 was 29.0 kD, and the theoretical isoelec- tric point （pl） was 8.25. The result of phylogenetic analysis showed that IGFBP-7 gone in Liangshan semi-fine wool sheep exhibited close phylogenetic relationships with bovine, goat and other mammals, and distant phylogenetic relationships with Danio rerio and Haliotis diversicolor. IGFBP-7 gene had uniformly distributed hy- drophobic and hydrophilic regions, harboring one signal peptide, two transmembrane regions, 16 phosphorylation sites, four N-glycosylation sites and one O-glyco- sylation site. The result of secondary structure analysis showed that the random coil, or-helix and β-sheet regions accounted for 64.89%, 19.86% and 15.25%, respectively. The result of tertiary structure analysis showed that IGFBP-7 harbors an IGFBP_N domain and an Ig-like domain. This study provided scientific basis for further investigating the function of IGFBP-7 gene in sheep.