Research progress in correlation between insulin-like growth factor 2 mRNA binding protein 2 gene and type 2 diabetes mellitus / 中国生物制品学杂志

@#Type 2 diabetes mellitus(T2DM)was a chronic,non-communicable disease with a combination of multiple genetic and environmental factors,of which the main characteristics included insufficient insulin secretion and insulin resistance.Insulin-like growth factor 2 mRNA binding protein 2(IGF2BP2/IMP2),an important insulin secretion-related protein in human body,is mainly expressed in tissues and cells such as pancreas,fat and intestine.It has been confirmed that IGF2BP2 can down-regulate the expression of IGF2 and the function damage of the related islet β cell is an important cause of T2DM and vascular complications.Therefore,IGF2BP2 gene can be used as an important predictor for diabetes mellitus risk.This paper reviews the correlation between IGF2BP2 gene and T2DM.

Role and mechanism of IGF2BP3 in malignant transformation of human gastric epithelial cells induced by MNNG / 环境与职业医学

Background N6-methyladenosine (m6A) RNA methylation may play an important role in the process of malignant transformation of cells induced by environmental carcinogens. However, the specific roles and mechanisms need to be further explored. Objective To explore the role and mechanism of m6A binding protein insulin-like growth factor 2 mRNA-binding protein 3 (IGF2BP3) in the malignant transformation of human gastric mucosal epithelial cells GES-1 induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). Methods Based on the GES-1 malignant transformation cells MC-30, a stable knockdown IGF2BP3 MC-30 cell line (MC30-shIGF2BP3, abbreviated as MC30-shI3) was constructed by lentiviral transfection technology, and a negative control group (MC30-NC) was also prepared. Real-time quantitative polymerase chain reaction (qRT-PCR) and Western blotting were applied to detect the mRNA expression and protein levels of IGF2BP3. RNA binding protein immunoprecipitation (RIP-qPCR) was used to examine the combination between IGF2BP3 protein and MYC mRNA in malignant cells MC-30. Furthermore, the stability of MYC mRNA was detected by actinomycin D assay. CCK-8 and Transwell respectively were employed to detect cell proliferation, migration, and invasion. Western blotting was applied to detect the expression of EMT markers (N-cadherin, Vimentin, α-SMA, and Snail). The role of the downstream target gene MYC was further elucidated by a rescue assay in MC30-shI3 cells transfected with a plasmid overexpressing MYC to observe changes in cellular phenotypes (proliferation, migration, invasion) and expression of key EMT proteins. Results Compared with the control group, the expression of IGF2BP3 mRNA was up-regulated after 5, 10, 20, and 40 μmol·L−1 MNNG infection of GES-1 cells (P<0.05). After 20 μmol·L−1 MNNG infection, the expression level of IGF2BP3 mRNA increased with prolongation of exposure time (P<0.05). Compared with the control group, the mRNA and protein expression levels of IGF2BP3 were up-regulated in the 10th, 20th, and 30th generations of 5 μmol·L−1 MNNG malignant transformation (P<0.05). The results of qRT-PCR and Western blotting showed that, compared with the MC30-NC group, the IGF2BP3 and MYC mRNA expression and protein expression decreased in the MC30-shI3 group (P<0.01). The CCK8 and transwell assay results showed that, compared with the MC30-NC group, the cell proliferation, migration, and invasion abilities significantly reduced in the MC30-shI3 group (P<0.01). The results of the Western blotting showed that, compared with the MC30-NC group, the protein levels of EMT markers N-cadherin, Vimentin, α-SMA, and Snail decreased in the MC30-shI3 group (P<0.01). The results of RIP-qPCR showed that, compared with the IgG group, the mRNA level was higher for the enriched MYC in the IGF2BP3 group (P<0.01); the results of the actinomycin D assay showed that, compared with the MC30-NC group, the stability of MYC mRNA significantly reduced in the MC30-shI3 group (P<0.01). While the rescue experiment showed that, compared with the IGF2BP3 knock-down+vector group, the MYC protein level significantly increased in the IGF2BP3 knock-down + MYC over-expression group (P<0.01), the proliferation, migration, and invasion abilities significantly enhanced (P<0.01), and the EMT key proteins (N-cadherin, Vimentin, α-SMA, Snail) increased in the MC30-shI3+MYC group (P<0.01). Conclusion Exposure to MNNG could result in up-regulation of IGF2BP3 expression in GES-1 cells. IGF2BP3 may enhance the proliferation, migration, and invasion of malignantly transformed human gastric epithelial cells by binding to MYC mRNA and increasing its stability and expression level and thus promoting the EMT process, which in turn affects the progression of malignant transformation.

The mechanism of enriched environment repairing the learning and memory impairment in offspring of prenatal stress by regulating the expression of activity-regulated cytoskeletal-associated and insulin-like growth factor-2 in hippocampus

BACKGROUND@#Prenatal stress can cause neurobiological and behavioral defects in offspring; environmental factors play a crucial role in regulating the development of brain and behavioral; this study was designed to test and verify whether an enriched environment can repair learning and memory impairment in offspring rats induced by prenatal stress and to explore its mechanism involving the expression of insulin-like growth factor-2 (IGF-2) and activity-regulated cytoskeletal-associated protein (Arc) in the hippocampus of the offspring.@*METHODS@#Rats were selected to establish a chronic unpredictable mild stress (CUMS) model during pregnancy. Offspring were weaned on 21st day and housed under either standard or an enriched environment. The learning and memory ability were tested using Morris water maze and Y-maze. The expression of IGF-2 and Arc mRNA and protein were respectively measured by using RT-PCR and Western blotting.@*RESULTS@#There was an elevation in the plasma corticosterone level of rat model of maternal chronic stress during pregnancy. Maternal stress's offspring exposed to an enriched environment could decrease their plasma corticosterone level and improve their weight. The offspring of maternal stress during pregnancy exhibited abnormalities in Morris water maze and Y-maze, which were improved in an enriched environment. The expression of IGF-2, Arc mRNA, and protein in offspring of maternal stress during pregnancy was boosted and some relationships existed between these parameters after being exposed enriched environment.@*CONCLUSIONS@#The learning and memory impairment in offspring of prenatal stress can be rectified by the enriched environment, the mechanism of which is related to the decreasing plasma corticosterone and increasing hippocampal IGF-2 and Arc of offspring rats following maternal chronic stress during pregnancy.

Epigenetic characterization of the H19/IGF2 locus in calf clones placenta / Caracterização epigenética do locus H19/IGF2 na placenta de bezerros clones

Somatic Cell Nuclear Transfer (SCNT-Cloning) is a promising technique in many areas and is based on genetically identical individuals. However, its efficiency is low. Studies suggest that the leading cause is inadequate epigenetic reprogramming. This study aimed to characterize the methylation pattern of the exon 10 regions of the IGF2 gene and the Imprinting Control Region (ICR) of the H19 gene in the placenta of cloned calves. For this study, female and male cloned calves presenting different phenotypes were used. Genomic DNA from these animals' placenta was isolated, then treated with sodium bisulfite and amplified to the ICR/H19 and IGF2 loci. PCR products were cloned into competent bacteria and finally sequenced. A significant difference was found between controls and clones with healthy phenotypes for the ICR/H19 region. In this region, controls showed a hemimethylated pattern, as predicted in the literature due to this region has an imprinted control, while clones were showed less methylated. For the IGF2, no significant differences were found between controls and clones. These results suggest that different genomic regions in the genome may be independently reprogrammed and that failures in reprogramming the DNA methylation patterns of imprinted genes may be one of the causes of the low efficiency of SCNT.(AU)

A Transferência Nuclear de Células Somáticas (TNCS-Clonagem) é uma técnica promissora em várias áreas, e se baseia na produção de indivíduos geneticamente idênticos. No entanto, sua eficiência é baixa. Estudos sugerem que a principal causa seja uma reprogramação epigenética inadequada. O objetivo desse trabalho é caracterizar o padrão de metilação da região éxon 10 do gene IGF2 e da Região Controladora de Imprinting (ICR) do gene H19 na placenta de bezerros clonados. Para a execução do trabalho foram selecionados clones bovinos fêmeas e machos, apresentando diferentes fenótipos. O DNA da placenta desses animais foi extraído, e em seguida foi tratado com bissulfito de sódio e amplificado para os loci ICR/H19 e IGF2. Os produtos da PCR foram clonados em bactérias competentes e, por fim, sequenciados. Foi encontrada uma diferença significativa entre os controles e os clones com fenótipos saudáveis para a região da ICR/H19. Nesta região, os controles tiveram um padrão hemimetilado, como previsto pela literatura, devido essa região ser imprinted. Enquanto os clones encontravam-se menos metilados. Para a região do éxon 10 do IGF2, não foi encontrada diferença significativa entre controles e clones. Estes resultados sugerem que as diferentes regiões do genoma podem se reprogramar independente umas das outras e que falhas na reprogramação do padrão de metilação do DNA de genes imprinted podem ser uma das causas da baixa eficiência da TNCS.(AU)

Association between metabolic syndrome and euthyroid nodular goiter: a case-control study / Asociación entre el síndrome metabólico y el bocio nodular eutiroideo: un estudio de casos y controles

Abstract Background: Metabolic syndrome is a cluster of metabolic abnormalities and abdominal obesity; its pathophysiologic basis, insulin resistance, has been shown to act as agent in thyroid cell proliferation. Few studies analyze the relationship between metabolic syndrome and thyroid nodular disease, with a substantial knowledge gap. Objective: Determine the association between metabolic syndrome and nodular thyroid disease in a region with adequate iodine intake. Methods: Case-control study. A total of 182 patients referred to radiology to undergo thyroid ultrasonography due to suspicion of thyroid disease. Cases had at least one thyroid nodule greater than 3 mm (n= 91). Controls did not have evidence of thyroid nodules (n= 91). Results: Bivariate analysis showed a significant association between metabolic syndrome and the presence of thyroid nodule (OR 2.56, 95% CI: 1.41-4.66, p <0.05). Low levels of HDL (OR 2.81, 95% CI: 1.54-5.12, p <0.05) and impaired fasting glucose (OR 2.05, 95%CI 1.10 to 3.78, p <0.05) were significantly associated with the presence of thyroid nodule, independent of the presence of metabolic syndrome. Multivariate analysis maintained the association between metabolic syndrome and thyroid nodule with an OR of 2.96 (95%CI 1.47 to 5.95, p <0.05); similarly, the associations of low levels of HDL (OR 2.77, 95%CI 1.44 to 5.3, p <0.05) and impaired fasting glucose (OR 2.23, 95%CI 1.14 to 4.34, p<0.05) with thyroid nodule remained significant. Conclusion: The thyroid nodular disease is associated with increased risk of metabolic syndrome, specifically decreased HDL and impaired fasting glucose levels were the factors that increased association was found.

Resumen Antecedentes: el síndrome metabólico es un conjunto de anormalidades metabólicas y obesidad abdominal; Se ha demostrado que su base fisiopatológica, la resistencia a la insulina, actúa como agente en la proliferación de las células tiroideas. Pocos estudios analizan la relación entre el síndrome metabólico y la enfermedad nodular tiroidea, con una brecha de conocimiento sustancial. Objetivo: determinar la asociación entre el síndrome metabólico y la enfermedad tiroidea nodular en una región con una ingesta adecuada de yodo. Métodos: estudio de casos y controles. Un total de 182 pacientes remitidos a radiología para someterse a una ecografía tiroidea debido a la sospecha de enfermedad tiroidea. Los casos tenían al menos un nódulo tiroideo mayor de 3 mm (n = 91). Los controles no tenían evidencia de nódulos tiroideos (n = 91). Resultados: El análisis bivariado mostró una asociación significativa entre el síndrome metabólico y la presencia de nódulo tiroideo (OR 2.56, IC 95%: 1.41-4.66, p <0.05). Los niveles bajos de HDL (OR 2.81, IC 95%: 1.54-5.12, p <0.05) y glucosa en ayunas alterada (OR 2.05, IC 95% 1.10 a 3.78, p <0.05) se asociaron significativamente con la presencia de nódulo tiroideo, independiente de la presencia de síndrome metabólico. El análisis multivariado mantuvo la asociación entre el síndrome metabólico y el nódulo tiroideo con un OR de 2.96 (IC 95% 1.47 a 5.95, p <0.05); de manera similar, las asociaciones de niveles bajos de HDL (OR 2.77, IC 95% 1.44 a 5.3, p <0.05) y glucosa en ayunas alterada (OR 2.23, IC 95% 1.14 a 4.34, p <0.05) con nódulo tiroideo permanecieron significativas. Conclusión: la enfermedad nodular tiroidea se asocia con un mayor riesgo de síndrome metabólico, específicamente la disminución de HDL y los niveles de glucosa en ayunas alterados fueron los factores que aumentaron la asociación.

Marcadores de selección en cerdos Pampa Rocha: comparación con razas autóctonas de España y Portugal / Selection markers in Pampa Rocha piqs: a comparison with autochthonous breeds of Spain and Portugal

RESUMEN Objetivo. El análisis de marcadores de selección permite obtener datos de la vida evolutiva de una raza o línea y permite también evaluar la conveniencia o no de su uso en programas de mejora genética. Hemos evaluado SNPs en cuatro genes (IGF2, MC4R, PRKAG3 y PEPCK-C), que tienen importantes efectos fenotípicos, en cerdos de la raza Pampa Rocha, una raza criolla, y hemos comparado sus frecuencias alélicas con cerdos de diversas razas autóctonas y líneas de España y Portugal no sometidas a selección así como con jabalíes y cerdos de la raza Piétrain. Materiales y métodos. Los SNPs fueron analizados mediante diversas técnicas de RT-PCR. Resultados. Los resultados de los análisis muestran una similitud de frecuencias alélicas entre los cerdos de la raza Pampa Rocha y los cerdos autóctonos de la península ibérica sobre todo en el gen IGF2 y, en menor medida en el gen PEPCK-C. Sin embargo difieren considerablemente en el caso del marcador MC4R y, también en menor medida, en PRKAG3. En el trabajo se discute el uso potencial de los resultados obtenidos para orientar la selección genética de cerdos de la raza Pampa Rocha. Conclusiones. Nuestros resultados demuestran la peculiaridad de la raza Pampa Rocha con respecto a los marcadores estudiados.

ABSTRACT Objective. The analysis of selection markers allows to obtain information about the evolutive story of a particular breed or line and allows also to evaluate the usefulness of those markers for breeding programs. We have analyzed SNPs in four genes of the creole pig breed Pampa Rocha and we have compared their allelic frequencies with the allelic frequencies of diverse autochthonous breeds of Spain and Portugal and also with Piétrain pigs and wild boars. Materials and methods. The SNPs were analyzed using diverse RT-PCR methods. Results. The results of the analysis show that Pampa Rocha pigs have similar allelic frequencies with the autochthonous breeds of Spain and Portugal especially in the case of IGF2 and also, but not so coincident, in the case of PEPCK-C. However, they differ considerably for MC4R, and also, but in a lower extent, for PRKAG3. We discuss in this work the usefulness of our results for breeding of Pampa Rocha pigs. Conclusions. Our results demonstrate the peculiarity of the Pampa Rocha breed regarding the markers studied.

Study of miR-18a/IGF-2/Akt pathway regulating the proliferation, migration and invasion of ovarian cancer cells / 中华内分泌外科杂志

Objective@#To study the role of miR-18a in regulating the proliferation, migration and invasion of ovarian cancer cells through IGF-2/Akt pathway.@*Methods@#Ovarian cancer SKOV3 cells were cultured and grouped. The negative control group were not transfected with the plasmid, the blank plasmid group were transfected with the blank pcDNA3.1 plasmid, the IGF-2 plasmid group was transfected with the pcDNA3.1 plasmid expressing IGF-2, and the IGF-2 plasmid+L294002 group was transfected with pcDNA3.1 plasmid expressing IGF-2 and treated with Akt inhibitor LY294002. The upstream miRNA of IGF-2 was analyzed by bioinformatics method and verified by the double luciferase reporter gene method. The effect of overexpressing miR-18a on the expression of IGF-2 was observed.@*Results@#Transfection of IGF-2 expression plasmid can increase the expression level of IGF-2 in cells and the content of IGF-2 in culture medium (P<0.05) . Cell proliferation in the IGF-2 plasmid group (0.93±0.22, F=9.629, P<0.05) , migration (80.22±13.28, F=12.689, P<0.05) , invasion (72.31±11.38, F=33.845, P<0.05) , the relative expression levels of p-PI3K (1.35±0.22, F=8.321, P<0.05) and p-Akt (0.94±0.18, F=9.612, P<0.05) . The proliferation, migration, invasion viability, in the viability and cells were significantly higher than those in the negative control group and the blank plasmid group. And relative expression levels of p-PI3K and p-Akt in the IGF-2 plasmid+L294002 group were significantly lower than those in the IGF-2 plasmid group (P<0.05) . IGF-2 is a target gene of miR-18a, and overexpression of miR-18a can down-regulate the expression level of IGF-2 (P<0.05) .@*Conclusion@#IGF-2 can promote the proliferation, migration, and invasion of ovarian cancer cells, and this promotion is related to the activation of the Akt pathway, and miR-18a may be an upstream regulatory molecule of IGF-2.

LncRNA MALAT1 promotes proliferation and metastasis of cervical cancer cell via regulating miR-124-3p/IGF2BP1 axis / 中国肿瘤生物治疗杂志

@# Objective: To investigate the mechanism of lncRNA MALAT1 modulating proliferation and metastasis of cervical cancer cells via regulating miR-124-3p/IGF2BP1 axis. Methods: A total of 45 cases of cervical cancer tissues and corresponding paracancerous tissues resected from patients, who were admitted to the Department of Obstetrics and Gynecology of Guiyang Maternal and Child Health Hospital during April 2014 and December 2017, were included in this study; in addition, cervical cancer cell lines SiHa, Caski, HeLa and C33awere also collected for this study. qPCR was applied to detect the expression of MALAT1 in cervical cancer tissues and cell lines. MALAT1-knockdown vectors, miR-124-3p inhibitors and IGF2BP1-overexpression vectors were constructed and used to transfect cervical cancer cells, respectively; the influence of MALAT1 or MALAT1 knockdown on cell proliferation, invasion and epithelial mesenchymal transition (EMT) via miR-124-3p/IGF2BP1 axis were determined by CCK-8 assay, Transwell assay, Wb and immunofluorescence, respectively. The interaction between MALAT1, miR-124-3p, and IGF2BP1 were verified by dual luciferase reporter gene assay. Results: MALAT1 was up-regulated in cervical cancer tissues and cell lines (P<0.05 or P<0.01). Meanwhile, MALAT1 knockdown remarkably inhibited proliferation, invasion and EMT of cervical cancer cells (P<0.05 or P<0.01). Moreover, dual-luciferase reporter gene assay showed that MALAT1 directly interacted with miR-124-3p and down-regulated its expression, while miR-1243p negatively regulated IGF2BP1 expression. Our experiment further validated that MALAT1 knockdown suppressed proliferative, invasion and EMT of cervical cancer cells via inducing the inhibitory effect of miR-124-3p on IGF2BP1 (P<0.05 or P<0.01). Conclusion: MALAT1 promotes the proliferation, invasion and EMT of cervical cancer cells by down-regulating miR-124-3p/IGF2BP1 axis, which provides potential molecular targets for early diagnosis or treatment of cervical cancer.

Study of miR-18a/IGF-2/Akt pathway regulating the proliferation, migration and invasion of ovarian cancer cells / 中华内分泌外科杂志

Objective To study the role of miR-18a in regulating the proliferation, migration and inva-sion of ovarian cancer cells through IGF-2/Akt pathway. Methods Ovarian cancer SKOV3 cells were cultured and grouped. The negative control group were not transfected with the plasmid, the blank plasmid group were transfected with the blank pcDNA3.1 plasmid, the IGF-2 plasmid group was transfected with the pcDNA3.1 plas-mid expressing IGF-2, and the IGF-2 plasmid+L294002 group was transfected with pcDNA3.1 plasmid expressing IGF-2 and treated with Akt inhibitor LY294002. The upstream miRNA of IGF-2 was analyzed by bioinformatics method and verified by the double luciferase reporter gene method. The effect of overexpressing miR-18a on the expression of IGF-2 was observed. Results Transfection of IGF-2 expression plasmid can increase the expres-sion level of IGF-2 in cells and the content of IGF-2 in culture medium (P<0.05). Cell proliferation in the IGF-2 plasmid group(0.93±0.22, F=9.629, P<0.05), migration(80.22±13.28, F=12.689, P<0.05), invasion(72.31±11.38, F=33.845, P<0.05), the relative expression levels of p-PI3K(1.35±0.22, F=8.321, P<0.05) and p-Akt(0.94±0.18, F=9.612, P<0.05). The proliferation, migration, invasion viability, in the viability and cells were significantly higher than those in the negative control group and the blank plasmid group. And relative expression levels of p-PI3K and p-Akt in the IGF-2 plasmid+L294002 group were significantly lower than those in the IGF-2 plasmid group(P<0.05). IGF-2 is a target gene of miR-18a, and overexpression of miR-18a can down-regulate the expres-sion level of IGF-2(P<0.05). Conclusion IGF-2 can promote the proliferation, migration, and invasion of ovari-an cancer cells, and this promotion is related to the activation of the Akt pathway, and miR-18a may be an up-stream regulatory molecule of IGF-2.

SIRT6 regulating IGF2 though recruiting mRNA binding protein IMP1 in 293T cells / 基础医学与临床

Objective To investigate the post-transcriptional regulation mechanism of insulin-like growth factor 2 (IGF2),which is an important tumor regulator,by SIRT6. Methods SIRT6 was overexpressed in 293T cells and IP was used to enrich SIRT6 and mass spectrometry(MS)was used to detect the protein that interacted with SIRT6. Western blot was used to validate the interacted protein and its acetylation/phosphorylation modification status. Results SIRT6 did not change the acetylation modification status of IMP1,but the phosphorylation status of IMP1 was elevated in the presence of SIRT6. Conclusions SIRT6 may regulate IGF2 though promoting the phosphoryla-tion of IMP1 in 293T cells.

Expressão do mRNA para IGF-2 em oócitos e células do cumulus extraídos de folículos antrais e pré-antrais de ovelhas nativas do Estado de Pernambuco / IGF-2 mRNA expression in oocytes and cumulus cells of antral and preantral native sheep follicles in Pernambuco State, Brazil

Objetivou-se avaliar a expressão do mRNA para o gene do fator de crescimento IGF-2 em oócitos e células do cumulus de ovelhas em diferentes estágios do desenvolvimento folicular. Os folículos classificados morfologicamente como antrais (terciários e pré-ovulatórios) foram aspirados manualmente para obtenção dos oócitos e células do cumulus. Os folículos pré-antrais (secundários) foram extraídos do córtex ovariano, por microdissecção, e os oócitos retirados. Nos dois grupos, os oócitos foram desnudados e agrupados em "pools" de dez células cada (Grupo A, n=10; Grupo B, n=10) e dez amostras com grupos de células do cumulus (Grupo A1, n=10, B1, n=10). O mRNA foi extraído e convertido em cDNA utilizando a técnica da RT-PCR, utilizando Oligo DT randômico para o mRNA. A análise da expressão confirmou a expressão gênica para IGF-2 nos grupos de oócitos e células do cumulus. Houve um aumento da expressão relativa do mRNA para IGF-2 nos grupos de oócitos durante a fase mais tardia do desenvolvimento folicular e as diferenças foram consideradas significantes (p<0,05). Não houve variação significante da expressão de IGF2 entre os grupos de células do cumulus. Conclui-se que o fator de crescimento IGF-2 tem níveis mais elevados de expressão em oócitos ovinos, na segunda fase do desenvolvimento folicular, mas expressão semelhante em células do cumulus durante as fases estudadas do desenvolvimento folicular.(AU)

The aim of this study was to analyze the mRNA expression of IGF-2 growth factor in oocytes and cumulus cells from native sheep follicles at different stages of follicular development. The classified morphologically as antral follicles (tertiary preovulatory) were aspirated manually to obtain the oocyte and the cumulus cells. The preantral follicles (secondary) were extracted from the ovarian cortex by microdissection, and oocytes were removed. In both groups, oocytes were denuded and grouped into "pools" of ten cells each (Group A, n=10, Group B, n=10) and ten samples with groups of cumulus cells (Group A1, n=10; B1, n=10). The mRNA was extracted and converted to cDNA using the RT-PCR technique. The expression analysis confirmed the expression of IGF-2 gene for groups of oocyte and the cumulus cells. There was an increase in the relative expression of mRNA for IGF-2 for groups of oocytes during the later stage of follicular development and differences were considered significant (p<0.05). There was no significant variation in the expression of IGF2 between groups of cumulus cells. It is concluded that the growth factor IGF-2 has higher levels of expression in sheep oocytes in the second stage of follicular development in the conditions adopted and similar expression in cumulus cells during various stages of follicular development.(AU)

DNA methylation modulates H19 and IGF2 expression in porcine female eye

Abstract The sexually dimorphic expression of H19/IGF2 is evolutionarily conserved. To investigate whether the expression of H19/IGF2 in the female porcine eye is sex-dependent, gene expression and methylation status were evaluated using quantitative real-time PCR (qPCR) and bisulfite sequencing PCR (BSP). We hypothesized that H19/IGF2 might exhibit a different DNA methylation status in the female eye. In order to evaluate our hypothesis, parthenogenetic (PA) cells were used for analysis by qPCR and BSP. Our results showed that H19 and IGF2 were over-expressed in the female eye compared with the male eye (3-fold and 2-fold, respectively). We observed a normal monoallelic methylation pattern for H19 differentially methylated regions (DMRs). Compared with H19 DMRs, IGF2 DMRs showed a different methylation pattern in the eye. Taken together, these results suggest that elevated expression of H19/IGF2 is caused by a specific chromatin structure that is regulated by the DNA methylation status of IGF2 DMRs in the female eye.

Neuroprotective effects and mechanisms of Cerebroprotein Hydrolysate for Injection (Ⅰ) on vascular dementia in rats / 药物评价研究

Objective To investigate the neuroprotective mechanisms of Cerebroprotein Hydrolysate for Injection (Ⅰ) on vascular dementia in rats.Method The rat vascular dementia model was prepared using an improved two-vessel occlusion method,and the common carotid artery was only isolated but not blocked in sham group.Rats were randomly divided into sham group,model group,Cerebroprotein Hydrolysate for Injection (Ⅰ) groups with low,medium and high dose (5,10,20 mg/kg) and Cerebroprotein Hydrolysate Injection group (Cerebrolysin,Positive drug,10 mg/kg).The drug was administered by iv injection of rat tail vein once a day for two weeks,while the same volume of saline was administered in sham and model group.At the end of administration,the plasma was collected through abdominal aorta to separate serum,and rat cortex was isolated to prepare homogenate.The levels of nerve growth factor (NGF) and insulin-like growth factor 2 (IGF-2) in serum and level of gamma-aminobutyric acid (GABA) in cortex were detected by ELISA.Level of glutamate (Glu) in cortex of VaD rats was detected by colorimetry.Results Compared with model group,levels of NGF and IGF-2 in the serum of VaD rats and level of GABA in cortex were significantly increased,while level of Glu in cortex was significantly decreased after administration of Cerebroprotein Hydrolysate for Injection (Ⅰ).The increased IGF-2 and GABA levels by Cerebroprotein Hydrolysate for Injection (Ⅰ) were significantly higher than that of Cerebrolysin at same dose.Conclusion The mechanisms underlying the increased leaming and memory ability of VaD rats by Cerebroprotein Hydrolysate for Injection (Ⅰ),are possibly related to the increased levels of NGF and IGF-2 in body and a regulation of the balance between excitatory and inhibitory amino acid neurotransmitters.

Neuroprotective effects and mechanisms of Cerebroprotein Hydrolysate for Injection (Ⅰ) on vascular dementia in rats / 药物评价研究

Objective To investigate the neuroprotective mechanisms of Cerebroprotein Hydrolysate for Injection (Ⅰ) on vascular dementia in rats.Method The rat vascular dementia model was prepared using an improved two-vessel occlusion method,and the common carotid artery was only isolated but not blocked in sham group.Rats were randomly divided into sham group,model group,Cerebroprotein Hydrolysate for Injection (Ⅰ) groups with low,medium and high dose (5,10,20 mg/kg) and Cerebroprotein Hydrolysate Injection group (Cerebrolysin,Positive drug,10 mg/kg).The drug was administered by iv injection of rat tail vein once a day for two weeks,while the same volume of saline was administered in sham and model group.At the end of administration,the plasma was collected through abdominal aorta to separate serum,and rat cortex was isolated to prepare homogenate.The levels of nerve growth factor (NGF) and insulin-like growth factor 2 (IGF-2) in serum and level of gamma-aminobutyric acid (GABA) in cortex were detected by ELISA.Level of glutamate (Glu) in cortex of VaD rats was detected by colorimetry.Results Compared with model group,levels of NGF and IGF-2 in the serum of VaD rats and level of GABA in cortex were significantly increased,while level of Glu in cortex was significantly decreased after administration of Cerebroprotein Hydrolysate for Injection (Ⅰ).The increased IGF-2 and GABA levels by Cerebroprotein Hydrolysate for Injection (Ⅰ) were significantly higher than that of Cerebrolysin at same dose.Conclusion The mechanisms underlying the increased leaming and memory ability of VaD rats by Cerebroprotein Hydrolysate for Injection (Ⅰ),are possibly related to the increased levels of NGF and IGF-2 in body and a regulation of the balance between excitatory and inhibitory amino acid neurotransmitters.

Expression of intronic miRNAs and their host gene Igf2 in a murine unilateral ureteral obstruction model

The objective of this study was to determine the expression of miR-483 and miR-483* and the relationship among them, their host gene (Igf2), and other cytokines in a murine model of renal fibrosis. The extent of renal fibrosis was visualized using Masson staining, and fibrosis was scored 3 days and 1 and 2 weeks after unilateral ureteral obstruction (UUO). Expression of miR-483, miR-483* and various cytokine mRNAs was detected by real-time polymerase chain reaction (PCR). Expression of miR-483 and miR-483* was significantly upregulated in the UUO model, particularly miR-483 expression was the greatest 2 weeks after surgery. Additionally, miR-483 and miR-483* expression negatively correlated with Bmp7 expression and positively correlated with Igf2, Tgfβ, Hgf, and Ctgf expression, as determined by Pearson's correlation analysis. Hgf expression significantly increased at 1 and 2 weeks after the surgery compared to the control group. This study showed that miR-483 and miR-483* expression was upregulated in a murine UUO model. These data suggest that miR-483 and miR-483* play a role in renal fibrosis and that miR-483* may interact with miR-483 in renal fibrosis. Thus, these miRNAs may play a role in the pathogenesis of renal fibrosis and coexpression of their host gene Igf2.

A hypomorphic Cbx3 allele causes prenatal growth restriction and perinatal energy homeostasis defects.

Mammals have three HP1 protein isotypes HP1β (CBX1), HP1γ (CBX3) and HP1α (CBX5) that are encoded by the corresponding genes Cbx1, Cbx3 and Cbx5. Recent work has shown that reduction of CBX3 protein in homozygotes for a hypomorphic allele (Cbx3hypo) causes a severe postnatal mortality with around 99% of the homozygotes dying before weaning. It is not known what the causes of the postnatal mortality are. Here we show that Cbx3hypo/hypo conceptuses are significantly reduced in size and the placentas exhibit a haplo-insufficiency. Late gestation Cbx3hypo/hypo placentas have reduced mRNA transcripts for genes involved in growth regulation, amino acid and glucose transport. Blood vessels within the Cbx3hypo/hypo placental labyrinth are narrower than wild-type. Newborn Cbx3hypo/hypo pups are hypoglycemic, the livers are depleted of glycogen reserves and there is almost complete loss of stored lipid in brown adipose tissue (BAT). There is a 10-fold reduction in expression of the BAT-specific Ucp1 gene, whose product is responsible for nonshivering themogenesis. We suggest that it is the small size of the Cbx3hypo/hypo neonates, a likely consequence of placental growth and transport defects, combined with a possible inability to thermoregulate that causes the severe postnatal mortality.

The relationship between serum insulin-like growth factor-2 levels and clinical characteristics in patients with schizophrenia / 中国神经精神疾病杂志

Objective To explore the change of serum insulin-like growth factor-2 (IGF-2) and its relationship with clinical characteristics in patients with schizophrenia. Methods Fifty-one schizophrenic patients were recruited in the present study and 50 healthy volunteers served as controls. The serum IGF-2 level was measured using enzyme linked immunosorbent assay (ELISA). Positive and Negative Syndrome Scale (PANSS) was used to evaluate the psychotic symp?toms of patients. Trail Making Test-A (TMTA), Digit-Symbol Coding Test (DSCT), Continuous Performance Test (CPT) and Stroop Color-Word Test (SCWT) were used to evaluate the cognitive function of both groups. Results There were sig?nificant differences in the results of TMTA, DSCT, CPT and SCWT between patient and control groups. The serum levels of IGF-2 were significantly lower in patients than that in controls [(202.7±40.7) ng/mL vs. (365.9±65.5) ng/mL, P0.05). Furthermore, significant correlations were found between the serum IGF-2 level and the negative symptom sub?scale of PANSS (r=-0.397, P=0.004), CPT score (r=0.378, P=0.006), SCWT-word number (r=0.289, P=0.040), SC? WT-color number (r=0.327, P=0.019) and SCWT-word/color number (r=0.386, P=0.005) in schizophrenic patients. Con?clusion The serum IGF-2 levels of patients with schizophrenia are significantly lower than that of healthy controls, and the IGF-2 level is associated with the severity of negative symptoms and cognitive impairments in patients, indicating that serum IGF-2 might be an indicator of the severity of schizophrenia.

The relationship between IGF2BP3 hypomethylation with colon tumor differentiation / 天津医药

Objective To investigate the relationship between IGF2BP3 hypomethylation with colon tumor differentia?tion. Methods Tissue samples from 41 colorectal cancer were collected from March 2011 to August 2011 in our hospital, among which 19 cases were well-differentiated adenocarcinoma, 12 cases were of moderately differentiated adenocarcinoma and 10 cases were of poorly differentiated adenocarcinoma. Meanwhile biopsy samples from 12 cases of colonic colitis were collected as a control. The expression of IGF2BP3 was assessed by immunohistochemistry and Western blot. An innovate of ELISA technique was used to examine global methylation levels while IGF2BP3 methylation level was tested by methylation specific PCR technique. Results The expressions of IGFBP3 are higher in all colon cancer groups than that in colitis (P<0.05). The expression of IGFBP3 in poorly differentiated adenocarcinoma is higher than that in all the other groups, but there is no significant difference between its expressions in the well differentiated group and in the moderately differentiated adeno?carcinoma group. The global DNA level and IGFBP3 methylation level of every colon cancer group is lower than those in coli?tis (P<0.05). Also, a tendency of decreasing global DNA and IGFBP3 methylation status was observed comparing well differ?entiated towards poorly differentiated carcinomas (P<0.05). Conclusion IGF2BP3 expression in colorectal cancer is asso?ciated with differentiation of colon cancer tissue. A lower global and IGF2BP3 DNA methylation suggest a worse differentia?tion of colon cancer. DNA hypomethylation may therefore play a regulatory role in the expression of IGF2BP3 in colon cancer tissue.

The value of detecting plasma microRNA-125a-3p,IGF-2 on monitoring invasion and metastasis in NSCLC / 重庆医学

Objective To investigate the value of detecting plasma microRNA‐125a‐3p(miRNA‐125a‐3p) ,IGF‐2 on monitoring invasion and metastasis in NSCLC ,and to study the correlation between miR‐125a‐3p and IGF‐2 .Methods miR‐125a‐3p transcripts of 20 controls ,73 NSCLC were performed in plasma by quantitative reverse transcription‐polymerase chain reaction(qRT‐PCR) and PCR data was analyzed by the 2‐ΔΔCT method .The expression of IGF‐2 in plasma was detected by ELISA .Results The expression of miR‐125a‐3p in stage Ⅲ /Ⅳ was lower than stage Ⅰ/Ⅱ and the controls(P=0 .001 ,P=0 .005) .There was no statistical differ‐ence between the stage Ⅰ /Ⅱ patients and the controls(P=0 .776) .The expression of miR‐125a‐3p was related with lymph node metastas ,lower expression in positive lymph node metastasis (P=0 .003) .The expression of IGF‐2 in stage Ⅰ /Ⅱ 、stage Ⅲ /Ⅳ was higher than the controls(P=0 .036 ,P=0 .011) .There was no statistical difference between the stageⅠ/Ⅱ and stage Ⅲ/Ⅳ (P=0 .451) . The expression of IGF‐2 was related with lymph node metastas ,higher expression in positive lymph node metastasis (P=0 .037) .The re‐sults showed a negative correlation between miR‐125a‐3p expression and IGF‐2 in plasma(r= -0 .280 ,P=0 .007) .Conclusion Low ex‐pression of miR‐125a‐3p and high expression of IGF‐2 in plasma may play a role in invasion and metastasis of NSCLC .miR‐125a‐3p may play a negative regulatory role on IGF‐2 .

Effects of parental folate deficiency on the folate content, global DNA methylation, and expressions of FRalpha, IGF-2 and IGF-1R in the postnatal rat liver

We examined the effect of parental folate deficiency on the folate content, global DNA methylation, folate receptor-alpha (FRalpha), insulin-like-growth factor-2 (IGF-2) and -1 receptor (IGF-1R) in the liver and plasma homocysteine in the postnatal rat. Male and female rats were randomly fed a folic acid-deficient (paternal folate-deficient, PD and maternal folate-deficient, MD), or folic acid-supplemented diet (paternal folate-supplemented, PS and maternal-folate-supplemented, MS) for four weeks. They were mated and grouped accordingly: PSxMS, PSxMD, PDxMS, and PDxMD. Pups were killed on day 21 of lactation. The hepatic folate content was markedly reduced in the PDxMD and PSxMD and PDxMS as compared with the PSxMS group. The hepatic global DNA methylation was decreased in the PDxMS and PSxMD groups as much as in the PDxMD group, and all the three groups were significantly lower as compared to the PSxMS group. There were no significant differences in the hepatic FRalpha, IGF-2 and IGF-1R expressions among the groups. Positive correlations were found between the hepatic folate content and global DNA methylation and protein expressions of FRalpha, IGF-2 and IGF-1R, whereas an inverse correlation was found between hepatic folate content and plasma homocysteine level in the 3-week-old rat pup. The results of this study show that both paternal and maternal folate deficiency at mating can influence the folate content and global DNA methylation in the postnatal rat liver.