BACKGROUNDPatients with single station mediastinal lymph node (N2) non-small cell lung cancer (NSCLC) have a better prognosis than those with multilevel N2. The molecular factors which are involved in disease progression remain largely unknown. The purpose of this study was to investigate gene expression differences between single station and multilevel N2 NSCLC and to identify the crucial molecular factors which are associated with progress and prognosis of stage N2 NSCLC.

METHODSGene expression analysis was performed using Agilent 4×44K Whole Human Genome Oligo Microarray on 10 freshfrozen lymph node tissue samples from single station N2 and paired multilevel N2 NSCLC patients. Real-time reverse transcription (RT)-PCR was used to validate the differential expression of 14 genes selected by cDNA microarray of which four were confirmed. Immunohistochemical staining for these validated genes was performed on formalin-fixed, paraffinembedded tissue samples from 130 cases of stage N2 NSCLC arranged in a high-density tissue microarray.

RESULTSWe identified a 14 gene expression signature by comparative analysis of gene expression. Expression of these genes strongly differed between single station and multilevel N2 NSCLC. Four genes (ADAM28, MUC4, CLDN1, and IGF2) correlated with the results of microarray and real-time RT-PCR analysis for the gene-expression data in samples from 56 NSCLC patients. Immunohistochemical staining for these genes in samples from 130 cases of stage N2 NSCLC demonstrated the expression of IGF2 and CLDN1 was negatively correlated with overall survival of stage N2 NSCLC.

CONCLUSIONSOur results suggest that the expression of CLDN1 and IGF2 indicate a poor prognosis in stage N2 NSCLC. Further, CLDN1 and IGF2 may provide potential targeting opportunities in future therapies.

Adult ; Aged ; Carcinoma, Non-Small-Cell Lung ; metabolism ; mortality ; pathology ; Claudin-1 ; analysis ; genetics ; Female ; Humans ; Immunohistochemistry ; Insulin-Like Growth Factor II ; analysis ; genetics ; Lung Neoplasms ; metabolism ; mortality ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis

PURPOSE: Insulin-like growth factors (IGFs) regulate a wide range of biological functions including cell proliferation, differentiation, and apoptosis through paracrine and autocrine mechanisms. Accordingly, the present study analyzed polymorphisms of IGF genes and their impact on the prognosis for patients with gastrointestinal stromal tumors (GISTs). METHODS: Two hundred-thirteen consecutive patients with GISTs who underwent curative surgery from 5 medical centers were enrolled in the present study. The genomic DNA was extracted from paraffin-embedded tumor tissue, and four IGF-1 (+2995C/A, +533C/T, IVS2-16540A/G, Ex4-177G/C) and one IGF-2 (IVS1+1280A/G) gene polymorphisms were determined using a Sequenom MassARRAY system. RESULTS: With a median follow-up of 18.4 months, the estimated 5-year relapse-free survival and overall survival rates were 69.9% and 86.7%, respectively. In a multivariate analysis including age, gender, primary site of disease, pathology, and risk stratification, no significant association was observed between the polymorphism of the IGF-1 and IGF-2 genes and survival. CONCLUSION: None of the five IGF-1 and IGF-2 gene polymorphisms investigated in this study was found to be an independent prognostic marker for Korean patients with surgically resected GIST. However, further studies on a larger scale are warranted to clarify the role of IGF-1 and IGF-2 gene polymorphisms as a prognostic biomarker for GIST patients.
Apoptosis ; Cell Proliferation ; DNA ; Follow-Up Studies ; Gastrointestinal Stromal Tumors ; Humans ; Insulin-Like Growth Factor I ; Insulin-Like Growth Factor II ; Multivariate Analysis ; Polymorphism, Single Nucleotide ; Prognosis ; Somatomedins ; Survival Rate

AIMUnderstanding the response of mesenchymal stem cells (MSCs) to mechanical strain and their consequent gene expression patterns will broaden our knowledge of the mechanobiology of distraction osteogenesis.

METHODOLOGYIn this study, a single period of cyclic mechanical stretch (0.5 Hz, 2,000 microepsilon) was performed on rat bone marrow MSCs. Cellular proliferation and alkaline phosphatase (ALP) activity was examined. The mRNA expression of six bone-related genes (Ets-1, bFGF, IGF-II, TGF-beta, Cbfa1 and ALP) was detected using real-time quantitative RT-PCR.

RESULTSThe results showed that mechanical strain can promote MSCs proliferation, increase ALP activity, and up-regulate the expression of these genes. A significant increase in Ets-1 expression was detected immediately after mechanical stimulation, but Cbfa1 expression became elevated later. The temporal expression pattern of ALP coincided perfectly with Cbfa1.

CONCLUSIONThe results of this study suggest that mechanical strain may act as a stimulator to induce differentiation of MSCs into osteoblasts, and that these bone-related genes may play different roles in the response of MSCs to mechanical stimulation.

Alkaline Phosphatase ; analysis ; Animals ; Antigens, Surface ; analysis ; Biomechanical Phenomena ; Bone Marrow Cells ; physiology ; Cell Culture Techniques ; Cell Differentiation ; physiology ; Cell Proliferation ; Core Binding Factor Alpha 1 Subunit ; analysis ; Fibroblast Growth Factor 2 ; analysis ; Insulin-Like Growth Factor II ; analysis ; Mesenchymal Stromal Cells ; physiology ; Osteoblasts ; physiology ; Osteogenesis, Distraction ; Pluripotent Stem Cells ; physiology ; Proto-Oncogene Protein c-ets-1 ; analysis ; Rats ; Stress, Mechanical ; Transforming Growth Factor beta ; analysis ; Up-Regulation ; physiology

OBJECTIVE: To explore the different effect of short 7-day gonadotropin releasing hormone agonist (GnRHa) protocol and GnRHa long protocol on the insulin-like growth factor II(IGF-II) and insulin-like growth factor binding protein-4 (IGFBP-4) levels in follicular fluid. METHODS: Eighty-eight infertile patients due to tubal factors were included in this study. They were randomly divided into a short 7-day GnRHa protocol group and a GnRHa long protocol group (n = 44). Follicular fluid was obtained from dominant follicles during oocyte retrieval. Levels of IGF-II and IGFBP-4 in the follicular fluid were detected by radioimmunoassay and enzyme-linked immunosorbent assay respectively. RESULTS: Duration of controlled ovarian stimulation was significantly shorter and the injected dosages of gonadotropin were significantly lower in the short 7-day protocol group. The differences in serum levels of estradiol and estradiol per mature follicle on the day of human chorionic gonadotropin injection between the two groups were not significant. The concentrations of IGF-II and IGFBP-4 in the follicular fluid of the short 7-day protocol group were significantly lower,while the difference of the ratio of IGF-II/IGFBP-4 between the two groups was not significant. Linear correlation analysis showed that IGF-II level in the follicular fluid was positively correlated to the total dose of gonadotropin. CONCLUSION The short 7-day and long GnRHa protocols may affect the concentrations of IGF-II and IGFBP-4 in the follicular fluid. However, changes of IGF-II and IGFBP-4 concentrations do not contribute to different clinical outcomes.
Embryo Transfer ; Female ; Fertilization in Vitro ; methods ; Follicular Fluid ; chemistry ; Gonadotropin-Releasing Hormone ; administration & dosage ; agonists ; Humans ; Insulin-Like Growth Factor Binding Protein 4 ; analysis ; Insulin-Like Growth Factor II ; analysis ; Ovulation Induction ; methods

OBJECTIVETo investigate relationship between expressions of hypoxia inducible factor-1alpha (HIF-1alpha) and insulin-like growth factor-II (IGF-II) as well as their correlation with angiogenesis and prognosis in gastric carcinoma.

METHODSHIF-1alpha and IGF-II mRNA expression were analyzed using in situ hybridization, microvessel density (MVD) was determined by anti-CD34 immunostaining in 118 cases gastric carcinoma.

RESULTSThe positive rates of HIF-1alpha mRNA and IGF-II mRNA were 49.2% and 47.4%, respectively. In stage T3-T4 cases, positive rates of HIF-1alpha and IGF-II mRNA expression, the frequencies of vessel invasion, lymph node and distant metastasis were significantly higher than those in stage T1-T2 cases. The mean MVD in stage T3-T4 tumors, vessel invasion, lymph node metastasis and distant metastasis were significantly more frequent than those in stage T1-T2 tumors. The mean MVD in tumors with positive HIF-1alpha and IGF-II mRNA expression was significantly higher than those in tumors without HIF-1alpha and IGF-II mRNA expression. The expression of HIF-1alpha was positively correlated with IGF-II mRNA. There were positive correlations between MVD and expression of HIF-1alpha and IGF-II mRNA, respectively. The mean survival time and 5-year survival rate in cases with positive HIF-1alpha and VEGF expression and MVD value >/= 41.5 were significantly shorter than those in cases with negative HIF-1alpha and VEGF expression and MVD value < 41.5.

CONCLUSIONSHIF-1alpha and IGF-II play an important role in tumor invasion and metastasis, especially in tumor angiogenesis. So test of the expression of HIF-1alpha and IGF-II may act as a useful index of treatment and prognosis.

Adult ; Aged ; Antigens, CD34 ; analysis ; Female ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Immunohistochemistry ; In Situ Hybridization ; Insulin-Like Growth Factor II ; metabolism ; Male ; Middle Aged ; Neovascularization, Pathologic ; genetics ; metabolism ; pathology ; Prognosis ; RNA, Messenger ; genetics ; metabolism ; Stomach Neoplasms ; blood supply ; genetics ; metabolism ; Survival Analysis

OBJECTIVE: Pathogenesis of the endometriosis is very complex and the etiology is still unclear. Our hypothesis is that there may be some difference in gene expression patterns between eutopic endometriums with or without endometriosis. In this study, we analyzed the difference of gene expression profile with cDNA microarray. METHODS: Endometrial tissues were gathered from patients with endometriosis or other benign gynecologic diseases. cDNA microarray technique was applied to screen the different gene expression profiles from early and late secretory phase endometria of those two groups. Each three mRNA samples isolated from early and late secretory phase of endometrial tissues of control were pooled and used as master controls and labeled with Cy3-dUTP. Then the differences of gene expression pattern were screened by comparing eutopic endometria with endometriosis, which were labeled with Cy5-dUTP. Fluorescent labeled probes were hybridized on a microarray of 4,800 human genes. RESULTS: Twelve genes were consistently overexpressed in the endometrium of endometriosis such as ATP synthase H transporting F1 (ATP5B), eukaryotic translation elongation factor 1, isocitrate dehydrogenase 1 (NADP+), mitochondrial ribosomal protein L3, ATP synthase H+ transporting (ATP5C1) and TNF alpha factor. Eleven genes were consistently down-regulated in the endometriosis samples. Many extracellular matrix protein genes (decorin, lumican, EGF-containing fibulin-like extracellular matrix protein 1, fibulin 5, and matrix Gla protein) and protease/protease inhibitors (serine proteinase inhibitor, matrix metalloproteinase 2, tissue inhibitor of metalloproteinase 1), and insulin like growth factor II associated protein were included. Expression patterns of selected eight genes from the cDNA microarray were confirmed by quantitative RT-PCR or real time RT-PCR. CONCLUSION: The result of this analysis supports the hypothesis that the endometrium from patients with endometriosis has distinct gene expression profile from control endometrium without endometriosis.
Adenosine Triphosphate ; Endometriosis* ; Endometrium* ; Extracellular Matrix ; Female ; Gene Expression* ; Genital Diseases, Female ; Humans ; Insulin-Like Growth Factor II ; Isocitrate Dehydrogenase ; Matrix Metalloproteinase 2 ; Oligonucleotide Array Sequence Analysis ; Peptide Elongation Factor 1 ; Ribosomal Proteins ; RNA, Messenger ; Transcriptome*

OBJECTIVEIntrauterine growth retardation (IUGR) may contribute to the disorder of development of fetal brains. L-arginine has been known to be effective in blood vessel distension and improving the blood circulation of placentas. Recent studies have shown that L-arginine can ameliorate the placental hypoxia and improve the development of fetus. This study aimed to explore the effects of L-arginine on the expression of insulin-like growth factor (IGF)-I, IGF-II, IGF binding protein-3(IGFBP3)and IGF-I mRNA in brains of IUGR rats and the possible mechanisms of L-arginine.

METHODSThirty-six pregnant rats were randomly assigned into four groups: Control, Model, Low dose L-arginine (100 mg/kg) and High-dose L-arginine (200 mg/kg L-arginine) groups (n=9 each). IUGR was induced by passive smoking in rats from the last three groups. L-arginine was administered for the last two groups between days 8 and 20 of gestation. On day 21 of gestation, the pup rats were delivered by cesarean section. The levels of IGF-I, IGF-II and IGFBP3 in the brains of pup rats were measured by enzyme-linked immunoadsordent assay (ELISA) and the expression of IGF-I mRNA was detected by fluorescence quantitative PCR (FQ-PCR).

RESULTSThe levels of IGF-I, IGF-II and IGF-I mRNA expression in the Model group were significantly lower than in the Control group, with the IGF-I levels of 0.789 +/- 0.062 ng/mg vs 0.947 +/- 0.042 ng/mg, the IGF-II levels of 0.270 +/- 0.020 ng/mg vs 0.374 +/- 0.015 ng/mg and the IGF-I mRNA expression of (13.12 +/- 1.39) x 10(4) cps/mug RNA vs (21.28 +/- 3.54) x 10(4) cps/mug RNA (P < 0.01). In contrast, the IGFBP3 levels in the Model group were significantly higher than in the Control group (0.253 +/- 0.011 ng/mg vs 0.089 +/- 0.015 ng/mg; P < 0.01). Low or high dose L-arginine treatment increased significantly the IGF-I levels from 0.789 +/- 0.062 ng/mg (Model group) to 0.937 +/- 0.067 ng/mg (low dose group) or 0.858 +/- 0.077 ng/mg (high dose group), the IGF-II levels from 0.270 +/- 0.020 ng/mg (Model group) to 0.318 +/- 0.018 ng/mg (low dose group) or 0.354 +/- 0.021 ng/mg (high dose group) and the IGF-I mRNA expression from (13.12 +/- 1.39) x 10(4) cps/mug RNA (Model group) to (19.24 +/- 2.48) x 10(4) cps/mug RNA (low dose group) or (17.35 +/- 2.30) x 10(4) cps/mug RNA (high dose group) (P < 0.01). The IGFBP3 levels were significantly reduced after low or high dose L-arginine treatment (0.132 +/- 0.006 ng/mg or 0.146 +/- 0.009 ng/mg) compared with those of the Model group (0.253 +/- 0.011 ng/mg) ( P < 0.01).

CONCLUSIONSL-arginine can increase the levels of IGF-I and IGF-II and the IGF-I mRNA expression, and decrease the IGFBP3 level in the brain of rats with IUGR induced by passive smoking, thereby offering protective effects against IUGR.

Animals ; Arginine ; pharmacology ; therapeutic use ; Female ; Fetal Growth Retardation ; metabolism ; prevention & control ; Insulin-Like Growth Factor Binding Protein 3 ; analysis ; Insulin-Like Growth Factor I ; analysis ; genetics ; Insulin-Like Growth Factor II ; analysis ; Male ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar

OBJECTIVEMany studies have demonstrated that low levels of insulin-like growth factor-I (IGF-I) may be associated with the hypoxic-ischemic brain damage (HIBD) and that IGF-I has a neuroprotective effect. The role of IGF-II, a structurally and functionally homologous polypeptide with IGF-I, is unclear in HIBD. This study was designed to observe the changes of serum and cerebrospinal fluid (CSF) IGF-II levels in neonates with hypoxic-ischemic encephalopathy (HIE) and to investigate its effects on HIE.

METHODSSerum and CSF IGF-II levels in 41 neonates with HIE were measured by radioimmunoassay in the acute phase (postnatal age 12-24 hrs) and the convalescence phase (postnatal age 10-12 days). The 41 HIE neonates included 10 cases of mild, 12 moderate, and 19 severe HIE. Serum samples of 10 normal neonates were used as controls.

RESULTSIn the acute phase, serum IGF-II levels in the Mild HIE group (203.28 +/- 40.09 ng/mL) and the Moderate HIE group (192.33 +/- 39.66 ng/mL) were not significantly reduced, but were obviously reduced in the Severe HIE group (116.72 +/- 39.50 ng/mL) compared with normal controls (229.38 +/- 43.39 ng/mL) (P<0.01). During the convalescence phase, serum IGF-II levels in the Mild HIE group (285.53 +/- 49.44 ng/mL) and in the Moderate HIE group (278.69 +/- 51.34 ng/mL) increased significantly (P < 0.01); CSF IGF-II levels increased in the Mild HIE group from 27.23 +/- 7.82 ng/mL (acute phase) to 81.58 +/- 9.77 ng/mL (convalescence phase) (P < 0.01) and also increased in the Moderate HIE group from 23.43 +/- 7.79 ng/mL (acute phase) to 78.48 +/- 10.44 ng/mL (convalescence phase) (P < 0.01). The patients from the severe HIE group whose neurological symptoms or signs were improved in the convalescence showed higher serum and CSF IGF-II levels than in the acute phase (254.08 +/- 48.50 ng/mL vs 122.21 +/- 46.26 ng/mL; 69.42 +/- 10.20 ng/mL vs 15.05 +/- 7.03 ng/mL; P < 0.01). A positive correlation was found between the serum and CSF IGF-II levels in the HIE group (r=0.69, P < 0.01).

CONCLUSIONSIGF-II levels in serum and CSF are associated with the pathogenesis and the prognosis of neonatal HIE.

Female ; Humans ; Hypoxia-Ischemia, Brain ; metabolism ; Infant, Newborn ; Insulin-Like Growth Factor II ; analysis ; cerebrospinal fluid ; Male

Adult ; Base Sequence ; Carcinoma, Hepatocellular ; metabolism ; Female ; Humans ; Insulin-Like Growth Factor II ; biosynthesis ; genetics ; Liver Neoplasms ; metabolism ; Male ; Middle Aged ; Molecular Sequence Data ; RNA, Messenger ; biosynthesis ; genetics ; Sequence Analysis, DNA

Biomarkers ; blood ; Biomarkers, Tumor ; blood ; Carcinoma, Hepatocellular ; blood ; diagnosis ; genetics ; Early Diagnosis ; Humans ; Insulin-Like Growth Factor II ; analysis ; Liver Neoplasms ; blood ; diagnosis ; genetics ; Protein Precursors ; blood ; Prothrombin ; RNA, Messenger ; analysis ; alpha-Fetoproteins ; metabolism