Molecular cloning of a novel secreted peptide, INM02, and regulation of its expression by glucose

Wang, Xuanchun; Gong, Wei; Yu, Li; Yang, Zhihong; Zhou, Wenbai; Wang, Mei; Yang, Zhen; Wen, Jie; Hu, Renming

doi:10.1677/joe-09-0086

Cited by 11 publications

(20 citation statements)

References 19 publications

(14 reference statements)

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“…Use of those tissues was approved by the Medical Ethics Committee at Huashan Hospital, Fudan University. Northern blotting was performed using a kit (non-isotopic digoxigenin Northern Starter; Roche Diagnostics) as described previously [29].…”

Section: Methodsmentioning

confidence: 99%

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Glucose metabolism-related protein 1 (GMRP1) regulates pancreatic beta cell proliferation and apoptosis via activation of Akt signalling pathway in rats and mice

et al. 2011

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Aims/hypothesis We attempted to elucidate the impacts on and possible mechanisms by which glucose metabolismrelated protein 1 (GMRP1) affects beta cell survival. Methods Adenovirus-mediated GMRP1 overproduction and siRNA-mediated knockdown were performed in INS-1E cells and rat islets, after which cell proliferation or apoptosis were determined, and phosphorylation of Akt and BCL2-associated agonist of cell death (BAD) investigated. INS-1E cells and rat islets were cultured at 5.6 (low) or 25 mmol/l (high) glucose for 24 or 48 h, and cell proliferation or apoptosis and GMRP1 levels were investigated. INS-1E cells were treated for 24 h with 0, 10, 50 and 100 nmol/l insulin, and GMRP1 levels were determined. After INS-1E cells were transfected with siRNA for 72 h, high glucose-induced cell proliferation and insulinstimulated Akt phosphorylation were investigated. Glucose-infused rat models were established and beta cell proliferation and mass were evaluated. Levels of GMRP1, and phosphorylation of Akt and BAD were determined in glucose-infused islets. The GMRP1-mediated Akt pathway was also investigated in db/db mice. Results Overproduction of GMRP1 promoted beta cell proliferation via increased phosphorylation of Akt. Knockdown of Gmrp1 (also known as Btbd10) reduced phosphorylation of Akt with enhanced beta cell apoptosis. High glucose increased GMRP1 levels and cell proliferation in INS-1E cells and islet cells. Knockdown of Gmrp1 decreased high glucose-induced cell proliferation and insulin-stimulated Akt phosphorylation. Increased GMRP1 levels were involved in the enhancement of beta cell proliferation and mass in glucose-infused islets. Decreased GMRP1 levels may participate in beta cell apoptosis of db/db mice. Conclusions/interpretation GMRP1 regulates pancreatic beta cell proliferation and apoptosis via activation of Akt signalling pathway.

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Section: Methodsmentioning

confidence: 99%

“…The recombinant pET32-GMRP1 vector was transformed into E. coli strain BL21. Recombinant GMRP1 protein was produced and purified, and polyclonal antibody against human GMRP1 generated by immunising rabbits as described previously [29].…”

Section: Methodsmentioning

confidence: 99%

Glucose metabolism-related protein 1 (GMRP1) regulates pancreatic beta cell proliferation and apoptosis via activation of Akt signalling pathway in rats and mice

et al. 2011

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“…We, as well as others, have previously suggested that elevated glucose levels can cause altered gene expression in vascular and islets cells (14)(15)(16)(17) (18). He also reported that these genes were differentially regulated by high (22 mM) compared to low glucose (5.5 mM) concentrations.…”

Section: Introductionmentioning

confidence: 64%

Identification of the novel protein FAM172A, and its up-regulation by high glucose in human aortic smooth muscle cells

Dong²,

Leong³

et al. 2010

Int J Mol Med

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Abstract. The family with sequence similarity 172, member A (FAM172A) is a hypothetical protein. We recently cloned the FAM172A gene from normal human aortic tissues. In a previous study we also showed that the FAM172A gene was up-regulated by high glucose levels in macrophages. In the present study, we further identified the FAM172A protein at the level of translation and studied the effects of high glucose levels on its expression in human aortic smooth muscle cells. The FAM172A gene was subcloned into the eukaryotic expression vectors, PDC315 and pEGFP-N2. The cloned sequence shows an open reading frame of 1251 nucleotides encoding a protein of 416 amino acids. We further expressed the recombinant FAM172A protein and generated rabbit anti-human FAM172A polyclonal antibodies. The FAM172A protein was identified for the first time at the translation level by Western blot analysis. Western blotting also demonstrated that the FAM172A protein could be detected in human aortic endothelial, human aortic smooth muscle cells and THP-1-derived macrophages, the highest expression being observed in the human aortic smooth muscle cells. By a combination of bioinformatics and confocal laser scanning microscopy, we found that the FAM172A protein in HEK293 cells, was mainly located in the nucleus, and that there was an Arb2 conserved domain in the FAM172A protein sequence. We also presented evidence that the FAM172 protein expression in human aortic smooth muscle cells was up-regulated by high glucose levels in a concentration-dependent and time-course manner. We speculated that as a novel protein, FAM172A could be involved in the pathogenesis of high glucose-induced vascular damage. IntroductionThe chronic complications of diabetes are generally divided into microvascular and macrovascular complications. The diabetic macrovascular disease or atherosclerosis and its complications, are the leading causes of morbidity and mortality in patients with type 2 diabetes mellitus (1,2). The histopathology of atherosclerosis in patients with diabetes is similar to that in patients without diabetes. However, compared to non-diabetic patients, atherosclerotic vascular damage in diabetic patients is more diffuse, more complicated and with clinically worse outcomes (3). Although insulin resistance, hypertension, dyslipidemia and other risk factors certainly contribute to the progression of diabetic macroangiopathy (3-8), the exact mechanisms of these pathological processes remain to be elucidated.Chronic hyperglycemia is a hallmark characteristic of diabetes and exerts its adverse effects on the cardiovascular system in many ways (7). There is universal agreement that hyperglycemia is a powerful and independent risk factor for diabetic cardiovascular complications (9-13). Therefore, hyperglycemia plays a crucial role in the pathogenesis of diabetic macrovascular complications.Glucose is one of the most important factors by which the expressions of genes in vascular or other cells are regulated. We, as well as others, have previously sugges...

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“…Consequently, h HSS1 defines a new class of secreted factors. Although little is known about h HSS1, there is evidence that h HSS1 is one of the glucose-responsive genes with both mRNA and protein secretion being regulated by glucose [ 1 ]. As such, it is speculated that h HSS1 could be associated with the functions of pancreatic islets, specifically beta-cells [ 1 ].…”

Section: Introductionmentioning

confidence: 99%

“…Although little is known about h HSS1, there is evidence that h HSS1 is one of the glucose-responsive genes with both mRNA and protein secretion being regulated by glucose [ 1 ]. As such, it is speculated that h HSS1 could be associated with the functions of pancreatic islets, specifically beta-cells [ 1 ]. Recently, h HSS1 was identified as endoplasmic reticulum (ER) membrane protein complex subunit 10 (EMC10), one of the components of ER associated degradation (ERAD), an ubiquitin and proteasome dependent process [ 2 ].…”

Section: Introductionmentioning

confidence: 99%

Human hematopoietic signal peptide-containing secreted 1 (hHSS1) modulates genes and pathways in glioma: implications for the regulation of tumorigenicity and angiogenesis

et al. 2014

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BackgroundHuman Hematopoietic Signal peptide-containing Secreted 1 (hHSS1) is a truly novel protein, defining a new class of secreted factors. We have previously reported that ectopic overexpression of hHSS1 has a negative modulatory effect on cell proliferation and tumorigenesis in glioblastoma model systems. Here we have used microarray analysis, screened glioblastoma samples in The Cancer Genome Atlas (TCGA), and studied the effects of hHSS1 on glioma-derived cells and endothelial cells to elucidate the molecular mechanisms underlying the anti-tumorigenic effects of hHSS1.MethodsGene expression profiling of human glioma U87 and A172 cells overexpressing hHSS1 was performed. Ingenuity® iReport™ and Ingenuity Pathway Analysis (IPA) were used to analyze the gene expression in the glioma cells. DNA content and cell cycle analysis were performed by FACS, while cell migration, cell invasion, and effects of hHSS1 on HUVEC tube formation were determined by transwell and matrigel assays. Correlation was made between hHSS1 expression and specific genes in glioblastoma samples in the TCGA database.ResultsWe have clarified the signaling and metabolic pathways (i.e. role of BRCA1 in DNA damage response), networks (i.e. cell cycle) and biological processes (i.e. cell division process of chromosomes) that result from hHSS1effects upon glioblastoma growth. U87-overexpressing hHSS1 significantly decreased the number of cells in the G0/G1 cell cycle phase, and significantly increased cells in the S and G2/M phases (P < 0.05). U87-overexpressing hHSS1 significantly lost their ability to migrate (P < 0.001) and to invade (P < 0.01) through matrigel matrix. hHSS1-overexpression significantly decreased migration of A172 cells (P < 0.001), inhibited A172 tumor-induced migration and invasion of HUVECs (P < 0.001), and significantly inhibited U87 tumor-induced invasion of HUVECs (P < 0.001). Purified hHSS1 protein inhibited HUVEC tube formation. TCGA database revealed significant correlation between hHSS1 and BRCA2 (r = −0.224, P < 0.0005), ADAMTS1 (r = −0.132, P <0.01) and endostatin (r = 0.141, P < 0.005).ConclusionshHSS1-overexpression modulates signaling pathways involved in tumorigenesis. hHSS1 inhibits glioma-induced cell cycle progression, cell migration, invasion and angiogenesis. Our data suggest that hHSS1 is a potential therapeutic for malignant glioblastoma possessing significant antitumor and anti-angiogenic activity.

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Molecular cloning of a novel secreted peptide, INM02, and regulation of its expression by glucose

Abstract: We report the identification of a novel secreted peptide, INM02.

Cited by 11 publications

References 19 publications

Glucose metabolism-related protein 1 (GMRP1) regulates pancreatic beta cell proliferation and apoptosis via activation of Akt signalling pathway in rats and mice

Glucose metabolism-related protein 1 (GMRP1) regulates pancreatic beta cell proliferation and apoptosis via activation of Akt signalling pathway in rats and mice

Identification of the novel protein FAM172A, and its up-regulation by high glucose in human aortic smooth muscle cells

Human hematopoietic signal peptide-containing secreted 1 (hHSS1) modulates genes and pathways in glioma: implications for the regulation of tumorigenicity and angiogenesis

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