Growth Inhibitor Gene Research Articles

Identification of novel polymorphism in mammary-derived growth inhibitor gene of water buffalo and its expression analysis in the mammary gland

Mammary-derived growth inhibitor (MDGI), a member of the lipophilic family of fatty acid-binding proteins, plays an important role in the development, regulation, and differentiation of the mammary gland. The aim of the study was to identify polymorphism in the MDGI gene and its expression analysis in the mammary gland at various stages of lactation, in Indian buffalo. Nucleotide sequence analysis of MDGI gene in different breeds of riverine and swamp buffaloes revealed a total of 16 polymorphic sites and one Indel. Different transcription factor binding sites were predicted for buffalo MDGI gene promoter sequence, using online tools and in-silico analysis indicating that the SNPs in this region can impact the gene expression regulation. Phylogenetic analysis exhibited the MDGI of buffalo being closer to other ruminants like cattle, yak, sheep, and goats. Further, the expression analysis revealed that buffalo MDGI being highly expressed in well-developed mammary glands of lactating buffalo as compared to involution/non-lactating and before functional development to start the milk production stage in heifers. Stage-specific variation in expression levels signifies the important functional role of the MDGI gene in mammary gland development and milk production in buffalo, an important dairy species in Southeast Asia.

Longer or shorter spines: Reciprocal trait evolution in stickleback via triallelic regulatory changes in Stanniocalcin2a

Vertebrates have repeatedly modified skeletal structures to adapt to their environments. The threespine stickleback is an excellent system for studying skeletal modifications, as different wild populations have either increased or decreased the lengths of their prominent dorsal and pelvic spines in different freshwater environments. Here we identify a regulatory locus that has a major morphological effect on the length of stickleback dorsal and pelvic spines, which we term Maser (major spine enhancer). Maser maps in a closely linked supergene complex that controls multiple armor, feeding, and behavioral traits on chromosome IV. Natural alleles in Maser are differentiated between marine and freshwater sticklebacks; however, alleles found among freshwater populations are also differentiated, with distinct alleles found in short- and long-spined freshwater populations. The distinct freshwater alleles either increase or decrease expression of the bone growth inhibitor gene Stanniocalcin2a in developing spines, providing a simple genetic mechanism for either increasing or decreasing spine lengths in natural populations. Genomic surveys suggest many recurrently differentiated loci in sticklebacks are similarly specialized into three or more distinct alleles, providing multiple ancient standing variants in particular genes that may contribute to a range of phenotypes in different environments.

Ultrasound therapy with optimal intensity facilitates peripheral nerve regeneration in rats through suppression of pro-inflammatory and nerve growth inhibitor gene expression.

BackgroundTherapeutic ultrasound (US) is a promising physical therapy modality for peripheral nerve regeneration. However, it is necessary to identify the most effective US parameters and clarify the underlying mechanisms before its clinical application. The intensity of US is one of the most important parameters. However, the optimum intensity for the promotion of peripheral nerve regeneration has yet to be determined.ObjectivesTo identify the optimum intensity of US necessary for the promotion of peripheral nerve regeneration after crush injuries in rats and to clarify the underlying mechanisms of US by mRNA expression analysis.MethodsWe inflicted sciatic nerve crush injuries on adult Lewis rats and performed ultrasound irradiation using 4 different US intensities: 0 (sham stimulation), 30, 140, and 250 mW/cm2 with frequency (5 days/week) and duration (5 min/day). We evaluated peripheral nerve regeneration by quantitative real-time PCR one week after injury. Histomorphometric analyses and motor function analysis were evaluated 3 weeks after injury.ResultsUS stimulation enhanced re-myelination as well as sprouting of axons, especially at an intensity of 140 mW/cm2. mRNA expression revealed that US suppressed the expression of the inflammatory cytokines TNF and IL-6 and the axonal growth inhibitors SEMA3A and GSK3β.ConclusionsAn intensity of 140 mW/cm2 was optimal to support regeneration of the sciatic nerve after a crush injury in rats by, in part, the suppression of pro-inflammatory and nerve growth inhibitor gene expression.

Exercise modulates the levels of growth inhibitor genes before and after multiple sclerosis

Multiple sclerosis (MS) is a neurodegenerative disease of the central nervous system. The animal model of MS, experimental autoimmune encephalomyelitis (EAE), is commonly used for studies of human inflammatory demyelinating diseases and has been shown to be suitable for studying the effects of exercise on MS pathophysiology. The present study was conducted to determine the impact of forced swimming and voluntary running wheel exercises before and after the induction of EAE on expression of Nogo-A, NgR, and ROCK genes in the brain tissue. A total of 96 C57BL/6 mice were randomly divided into two groups, namely exercises before (EXb, n = 48) and after (EXa, n = 48) induction of EAE. Each group was divided into four subgroups: Forced Swimming + EAE (n = 12), Voluntary Running Wheel + EAE (n = 12), NoEX-EAE (n = 12), and Control group (n = 12). Animals performed either swimming exercise for 30 min per day or running wheel for one hour per day, five days per week for four weeks. Results of Luxal Fast Blue (LFB) staining demonstrated that the degree of demyelination was significantly less in the experimental exercised compared to NoEX-EAE groups (P < .05). Amazingly, both modes of exercise reduced the severity of MS symptoms in mice exposed to swimming and wheel running, evaluated as body weight, clinical scores, degree of demyelination, and gene expressions, regardless of whether the exercise was performed before or after EAE induction.

Shared Molecular Features Linking Endometriosis and Obstetric Complications

Recent epidemiological research has shown the increased risk of adverse pregnancy outcomes in women with endometriosis compared with the general population. The aim of this review is to explore common pathophysiologic mechanisms between endometriosis and obstetric complications. A computerized literature search was performed to identify relevant studies. The search covered the period between January 2008 and October 2018. One of the potential mechanisms driving the initiation and progression of endometriosis is the accumulation of a variety of epigenetic changes in endometrial cells. Epigenetic control of gene expression which is considered to be responsible for the development of endometriosis is commonly seen in patients with preeclampsia, small for gestational age (SGA), or preterm birth. DLX5 and GATA3, paternally imprinted genes, and CDKN1C, a maternally imprinted gene, were aberrantly expressed in placenta tissues of the preeclampsia; CDKN1C, the growth inhibitor gene, was upregulated in human SGA placentas; and hypomethylation of PTGER2 would be associated with preterm birth. Preeclampsia, SGA, or preterm birth may share common epigenetic alterations with endometriosis, which raises the possibility that the occurrence of two conditions might be nonrandom. To date, however, there is a lack of evidence that links endometriosis and other obstetric complications, such as postpartum hemorrhage or placental abruption, at the epigenetic level. In conclusion, epigenetic changes may be a common hallmark of two conditions: endometriosis and obstetrical complications, such as preeclampsia, SGA, or preterm birth.

Nutrition, epigenetic markers and growth in preterm infants

Background and aim Maternal diet and early nutrition of newborns may affect the phenotype later in adulthood. Susceptibility of epigenetic mechanisms to the nutritional environment is a critical element in neonatal development. Epigenetic mechanisms could be considered as a bridge between environmental stimuli and long lasting phenotype. IC2, a key region on 11p15, is involved in the control of growth and regulates CDKN1C, PHLDA2 and KCNQ1, growth inhibitor genes. Our aim was to investigate the relationship between epigenetic markers, nutrition and postnatal growth. Methods We enrolled 37 newborns (gestational age at birth was <34 weeks) admitted to Neonatal Intensive Care Unit at University Hospital of Pisa. Results We observed a relationship between reduced protein and lipid intake and IC2 hypermethylation (p = .003 and p = .001 respectively) and we also investigated the correlation between growth pattern and IC2 methylation. Conclusion The reduced growth, in part related to a reduced intake of nutrients (lipids and proteins), might be due to IC2 hypermethylation, causing an increased expression of growth inhibitor genes. IC2 hypermethylation could be a marker of reduced infants’ growth and may guides us to nutritional interventional strategies for a precocious prevention of extrauterine growth restriction (EUGR).

A case-control study about the association between vascular endothelial growth inhibitor gene polymorphisms and breast cancer risk in female patients in Northeast China.

Objective The inhibition of the neovascularization in tumors is a potential therapeutic target of cancer. Vascular endothelial growth inhibitor (VEGI) is a member of the TNF superfamily which has the ability to suppress the formation of new vessels in tumors. In order to study the association between VEGI gene polymorphisms and breast cancer risk, a case-control study was conducted in Chinese Han women in Northeast China. Methods Our study involved 708 female breast cancer patients and 685 healthy volunteers. Four SNPs of VEGI gene were analyzed through the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The association between VEGI gene polymorphisms and breast cancer risk was analyzed in our study. The relation between VEGI gene variants and clinical features of breast cancer including lymph node (LN) metastasis, estrogen receptor (ER), progestrogen receptor (PR), tumor protein 53 (p53), human epidermal growth factor receptor 2 (Her-2) and triple negative (ER-/PR-/Her-2-) status was analyzed as well.Results We found that the CT genotype and T allele of rs6478106 were more frequent in patients than in controls. There was also a statistical difference in the distribution of Crs6478106Grs4263839 haplotype between patients and controls. In addition, SNP rs6478106 and rs4979462 were related with the Her-2 status. Conclusions Our results suggest that VEGI gene variants may be related to the breast cancer risk and the clinical features of breast cancer in Chinese Han women in Northeast China.

Inhibition of corneal neovascularization by vascular endothelia growth inhibitor gene.

To evaluate the effect of Effectene™ lipofectine mediated plasmids encoding human pcDNA4-vascular endothelia growth inhibitor (pcDNA(4)-VEGI) gene on corneal neovascularization (CNV). Forty New Zealand albino rabbits were sutured by 5-0 silk on the superior cornea to establish the animal model and divided into 4 random group, ten per each group: group A: transfected by pcDNA(4)-VEGI gene mediated by Effectene™ lipofectine transfection, group B: by Plasmid pcDNA(4), group C: by Effectene™, and group D: by normal saline. Length and area of CNV were measured under slit lamp every day after transfection, immunohistochemistry was used to detected the expression of VEGI protein in cornea at 3, 7, 14 and 21 days. Average occurrence of CNV in the pcDNA(4)-VEGI gene transfected group (group A) was 6.3 days, in plasmid pcDNA(4) control group (group B) was 3.1 days, in Effectene™ lipofectine control group (group C) was 3.2 days, in normal saline control group (group D) was 3.2 days. Differences between groups A and B, C, D were statistically significant (P<0.01), while differences in groups B, C and D were meaningless (P>0.05). Lenth and average area of CNV in each period in group A was meaningful different from that in groups B, C, and D (P<0.01), while differences in group B, C and D were meaningless (P>0.05). Immunohistochemistry result: VEGI positive cells could be seen in epithelium, stroma, endothelium and the cliff of CNV in group A at 3 days after transfection. VEGI cells changed with the decrease of CNV. None positive cells were in the control groups (groups B, C and D) all the time. Effectene™ lipofectine transfection technique can be effectively used in transfecting pcDNA(4)-VEGI gene into rabbit cornea and the lenth and areas of CNV can be inhibited by VEGI gene.

Inhibition of corneal neovascularization by vascular endothelia growth inhibitor gene.

To evaluate the effect of Effectene™ lipofectine mediated plasmids encoding human pcDNA(4)-vascular endothelia growth inhibitor (pcDNA(4)-VEGI) gene on corneal neovascularization (CNV). Forty New Zealand albino rabbits were sutured by 5-0 silk on the superior cornea to induce CNV and divided into 4 random teams, ten per each team: team A: transfected by pcDNA(4)-VEGI gene mediated by Effectene(™) lipofectine transfection; team B: by plasmid pcDNA(4); team C: by Effectene(™), and team D: by normal saline. Length and area of CNV were observed under slit lamp every day after transfection. Immunohistochemistry was performed to detect the expression of VEGI protein in corneas at day 3, 7, 14 and 21. 1) Average occurrence of CNV was 6.3 days in team A, 3.1 days in team B, 3.2 days in team C, and 3.2 days in team D. Difference was significant between A and other teams (P<0.01); 2) Length and average area of CNV in each period in team A was significantly different from those in team B, C and D (P<0.01); 3) VEGI expressions were observed in epithelium, stroma, endothelium and the cliff of CNV in team A at 3 days after transfection by immunohistochemical staining. None VEGI positive cells were found in the control teams (team B, C and D) all the time. Effectene(™) lipofectine transfection technique can effectively transfect pcDNA(4)-VEGI gene into rabbit cornea and the length and CNV areas can be inhibited by VEGI gene.

Inhibition of corneal neovascularization by vascular endothelial growth inhibitor gene

To evaluate the effect of Effectene lipofectene mediated plasmids encoding human pcDNA4-vascular endothelia growth inhibitor (pcDNA4-VEGI) gene on corneal neovascularization (CNV). It was an experimental study. Forty healthy New Zealand albino rabbits (40 eyes) were divided into 4 teams according to the random digits table: team A (10 rabbits) was the team which transfected by lipofectine mediated plasmids encoding human pcDNA4-vascular endothelia growth inhibitor (pcDNA4-VEGI) gene; team B (10 rabbits) was the team transfected by empty vector; team C (10 rabbits) was the team transfected by lipofectine; team D (10 rabbits) was the empty control group which was transfected by saline. (1) Human VEGI gene fragment was connected with expressional vector plasmid pcDNA4 to construct the pcDNA4-VEGI gene expression vector. Computer automatic sequence analysis was used to identify the gene. (2) New Zealand albino rabbits were sutured by 5 - 0 silk in the superior cornea to establish the animal model and were transfected by pcDNA4-VEGI gene mediated by Effectene lipofectene transfection. Length and area of CNV were measured by slit lamp every day after transfection, immunohistochemistry was used to detected the expression of VEGI protein in cornea at the time 3, 7, 14 and 21 d. (1) Computer automatic sequence analysis confirmed the correct recombination of pcDNA4-VEGI gene. (2) Average occurrence of CNV in the pcDNA4-VEGI gene transfected team was 6.3 days while that in the control teams were 3.1, 3.2, 3.2 days (F = 39.838, P < 0.01). Average length and area of CNV were also different in the VEGI team and the control teams (q between team A and B on average length of CNV was 17.386, 20.944 and 8.892 on 7, 14 and 21 d; q between team A and C on average length of CNV was 19.488, 19.795 and 7.483 on 7, 14 and 21 d; q between team A and C on average length of CNV was 19.583, 20.413 and 8.941 on 7, 14 and 21 d, and P were all < 0.01. And q between team A and B on average area of CNV was 30.238, 57.820 and 35.543 on 7, 14 and 21 d; q between team A and C on average area of CNV was 32.607, 57.843 and 36.653 on 7, 14 and 21 d; q between team A and C on average area of CNV was 33.873, 57.590 and 34.724 on 7, 14 and 21 d, and P were all < 0.01). Integrated epithelium, light stroma edema, less CNV and less inflammation could be seen in the gene-therapy team. And immunohistochemistry results showed VEGI positive cells in epithelium, stroma, endothelium and the cliff of CNV in the gene therapy team. PCR, enzyme cutting, recombination and other genetic techniques could be used to construct expressional pcDNA4-VEGI gene. Effectene lipofectene transfection technique could be effectively used in transfecting pcDNA4-VEGI gene into rabbit cornea to inhibit CNV.

Isolation and Sequence Characterization of Mammary Derived Growth Inhibitor Gene of Riverine Buffalo (Bubalus Bubalis)

In this study, attempts have been made to identify and characterize water buffalo (Bubalus bubalis) mammary derived growth inhibitor (MDGI) gene, isolated from a mammary gland cDNA library of lactating buffalo. The complete MDGI cDNA was of 698 nucleotides, consisting 61 nucleotides in 5′ UTR, coding region of 402 nucleotides, and 235 nucleotides representing the 3′ UTR. Comparison of nucleotide and deduced amino acid sequence data with that of MDGI//fatty acid binding protein (FABP) of other species shows three buffalo specific nucleotide changes while seven nucleotide changes were common to cattle and buffalo. Buffalo and cattle MDGI had 100% amino acid sequence similarity, which also shared three amino acid changes: 34 (Ala-Gly), 109 (Leu-Met), and 132 (Glu-Gln) as compared to other species. Comparison with FABPs reported from other cattle tissues revealed highest amino acid sequence similarity with FABP-heart (100%) and least with FABP-liver (20.5%). Phylogenetic analysis revealed cattle MDGI to be closest to buffalo, while mouse MDGI was distantly placed, whereas different tissue derived FABPs of cattle showed FABP-heart closest and FABP-epidermis most distantly placed from buffalo MDGI. This report also differs from the earlier findings that MDGI is intermediate of FABP-heart and adipose.

Characterization of cis-regulatory elements of the vascular endothelial growth inhibitor gene promoter

VEGI (vascular endothelial growth inhibitor), a member of the tumour necrosis factor superfamily, has been reported to inhibit endothelial cell proliferation, angiogenesis and tumour growth. We identified and cloned approx. 2.2 kb of the VEGI promoter from mouse cerebral endothelial cells. The promoter contained an atypical TATA-box-binding protein sequence TAAAAAA residing at -32/-26 relative to the transcription initiation site (+1), 83 bp upstream from the ATG start codon. To investigate critical sequences in the VEGI promoter, a series of deleted and truncated segments were constructed from a 2300 bp promoter construct (-2201/+96) linked to a luciferase reporter gene. Transient transfection of cerebral microvascular cells (bEND.3) and rat C6 glioma cells demonstrated that a 1700 bp deletion from the -2201 to -501 did not significantly affect promoter activity; however, a truncated construct (-501/+96) lacking the region between -312 and -57 resulted in nearly 90% loss of promoter activity. A consensus NF-kappaB (nuclear factor kappaB) and several SP1 (specificity protein-1)-binding sequences were identified within the deleted segment. Supershift analysis revealed that NF-kappaB subunits, p50 and p65, interacted with the VEGI promoter. Exposure of cerebral endothermic cells to the pro-inflammatory cytokine, tumour necrosis factor-alpha, increased VEGI mRNA levels and DNA-binding activities, whereas an NF-kappaB inhibitor attenuated this increase. In addition, p65 overexpression enhanced, whereas p50 overexpression decreased, the luciferase activity. Furthermore, mutation of the NF-kappaB DNA binding site blocked this p65- and tumour necrosis factor-alpha-induced luciferase activity. These findings suggest that the transcription factor NF-kappaB plays an important role in the regulation of VEGI expression.

Packaging and identification of recombinant adenovirus carrying endostatin-soluble vascular endothelial growth inhibitor gene

Packaging and identification of recombinant adenovirus carrying endostatin-soluble vascular endothelial growth inhibitor gene

Suppression of tumorigenicity in human ovarian carcinoma cell line SKOV-3 by microcell-mediated transfer of chromosome 11

To determine the pathogenic role of chromosomes 11 and 17 in the carcinogenesis of human ovarian cancers, neo R-tagged chromosome 11 or 17 was transferred from cell lines A9H11 or A9H17, respectively, into the ovarian carcinoma cell line SKOV-3 using microcell-mediated chromosome transfer. The chromosome transfer was verified by polymerase chain reaction detection of the neo R gene, fluorescence in situ hybridization detection of an extra chromosome 11, and microsatellite polymorphism detection of an exogeneous chromosome 11. Five SKOV-3/A9H11 hybrids and five SKOV-3/A9H17 hybrid clones were generated. For the chromosome 11 transfer, complete suppression of tumorigenicity was observed in four clones, (11)9-8 and 11(H)7-2, 11(H)8-3, and 11(H)7-2, 100 days post implantation. For the chromosome 17 transfer, no complete suppression of tumorigenicity was observed. However, an increased latency period ranging from 25 to 49 days in contrast to 7 days for the SKOV-3 parental line, and a significant reduction in tumor size was observed. There was no correlation between the in vitro growth rate and the tumorigenicity or length of latency period. Our results demonstrate functionally that chromosome 11 may carry a tumor suppressor gene(s) while chromosome 17 may carry a tumor growth-inhibitor gene(s) for the ovarian carcinoma cell line, SKOV-3.

Reduced growth rate and transformation phenotype of the prostate cancer cells by an interferon-inducible protein, p202.

Interferons (IFNs) can exert cytostatic and immunomodulatory effects on carcinoma cells. In particular, growth inhibition of human prostate carcinoma by IFNs has been demonstrated both in vitro and in vivo. p202 is a 52 kd nuclear phosphoprotein known to be induced by IFNs. In this report, we showed that the expression of p202 was associated with an anti-proliferative effect on human prostate cancer cells. More importantly, cells that expressed p202 showed reduced ability to grow in soft-agar, indicating a loss of transformation phenotype. Our data suggest that p202 is a growth inhibitor gene in prostate cancer cells and its expression may also suppress transformation phenotype of prostate cancer cells.

Suppression of mammary-derived growth inhibitor gene expression by growth hormone and insulin-like growth factor I.

When introduced into MCF-7 breast cancer cells, the mammary-derived growth inhibitor (MDGI) gene causes them to revert to a more normal behavior. MDGI is silenced in several human breast cancer cell lines and in most breast tumors. Antiestrogens (tamoxifen and ICI 182780), which are commonly used in breast cancer treatment, stabilize MDGI mRNA. Insulin-like growth factors (IGFs) are well characterized mitogenic and anti-apoptotic factors involved in mammary gland physiology. We demonstrate that MDGI gene expression was inversely correlated with IGF-II gene expression. In the mammary gland of growth hormone releasing hormone receptor mutant (Ghrhrlit/Ghrhrlit) mice, the MDGI gene was overexpressed. Administration of IGF-I or GH to Ghrhrlit/Ghrhrlit mice suppressed MDGI mRNA levels in a dose-dependent manner. Administration of the somatostatin analogue octreotide to pituitary intact rats in a manner previously shown to acutely suppress the GH/IGF-I axis, up-regulated mammary gland MDGI expression in a dose-dependent fashion. The data document a previously unrecognized role of IGF-I in the regulation of the tumor suppressor gene MDGI, in the mammary gland, and may aid in the design of new physiological approaches to breast cancer prevention and/or treatment.

An update on the cellular and molecular biology of brain tumors.

Patients with tumors of the central nervous system (CNS) remain difficult to treat despite recent advances in surgical, chemotherapeutic and radiotherapeutic techniques. A better understanding of the molecular and cellular biology of neoplasia is providing neuroscientists with a framework on which to devise novel therapies for these patients. It thus becomes imperative that neurologists and neurosurgeons be aware of these advances in basic science that may eventually have a positive impact on patient management. This paper reviews our present knowledge of the process of CNS oncogenesis and the roles that chemicals, viruses, oncogenes, growth inhibitor genes, and growth factors play in the process.