Specific effects of antitumor active norspermidine on the structure and function of DNA

Abstract We compared the effects of trivalent polyamines, spermidine (SPD) and norspermidine (NSPD), a chemical homologue of SPD, on the structure of DNA and gene expression. The chemical structures of SPD and NSPD are different only with the number of methylene groups between amine groups, [N-3-N-4-N] and [N-3-N-3-N], respectively. SPD plays vital roles in cell function and survival, including in mammals. On the other hand, NSPD has antitumor activity and is found in some species of plants, bacteria and algae, but not in humans. We found that both polyamines exhibit biphasic effect; enhancement and inhibition on in vitro gene expression, where SPD shows definitely higher potency in enhancement but NSPD causes stronger inhibition. Based on the results of AFM (atomic force microscopy) observations together with single DNA measurements with fluorescence microscopy, it becomes clear that SPD tends to align DNA orientation, whereas NSPD induces shrinkage with a greater potency. The measurement of binding equilibrium by NMR indicates that NSPD shows 4–5 times higher affinity to DNA than SPD. Our theoretical study with Monte Carlo simulation provides the insights into the underlying mechanism of the specific effect of NSPD on DNA.

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Circulating miRNA 887 is differentially expressed in ARDS and modulates endothelial function

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00494.2019 ◽

2020 ◽

Vol 318 (6) ◽

pp. L1261-L1269 ◽

Cited By ~ 1

Author(s):

Andrew J. Goodwin ◽

Pengfei Li ◽

Perry V. Halushka ◽

James A. Cook ◽

Aman S. Sumal ◽

...

Keyword(s):

Gene Expression ◽

Cell Function ◽

In Silico Analysis ◽

Leukocyte Migration ◽

Differentially Expressed ◽

Expression Of Genes ◽

Cell Gene Expression ◽

And Function ◽

The Impact

Circulating microRNAs (miRNAs) can be taken up by recipient cells and have been recently associated with the acute respiratory distress syndrome (ARDS). Their role in host predisposition to the syndrome is unknown. The objective of the study was to identify circulating miRNAs associated with the development of sepsis-related ARDS and examine their impact on endothelial cell gene expression and function. We determined miRNA levels in plasma collected from subjects during the first 24 h of admission to a tertiary intensive care unit for sepsis. A miRNA that was differentially expressed between subjects who did and did not develop ARDS was identified and was transfected into human pulmonary microvascular endothelial cells (HPMECs). RNA sequencing, in silico analysis, cytokine expression, and leukocyte migration assays were used to determine the impact of this miRNA on gene expression and cell function. In two cohorts, circulating miR-887-3p levels were elevated in septic patients who developed ARDS compared with those who did not. Transfection of miR-887-3p into HPMECs altered gene expression, including the upregulation of several genes previously associated with ARDS (e.g., CXCL10, CCL5, CX3CL1, VCAM1, CASP1, IL1B, IFNB, and TLR2), and activation of cellular pathways relevant to the response to infection. Functionally, miR-887-3p increased the endothelial release of chemokines and facilitated trans-endothelial leukocyte migration. Circulating miR-887-3p is associated with ARDS in critically ill patients with sepsis. In vitro, miR-887-3p regulates the expression of genes relevant to ARDS and neutrophil tracking. This miRNA may contribute to ARDS pathogenesis and could represent a novel therapeutic target.

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The Proliferation Enhancing Effects of Salidroside on Schwann Cells In Vitro

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2017/4673289 ◽

2017 ◽

Vol 2017 ◽

pp. 1-10 ◽

Cited By ~ 2

Author(s):

Hui Liu ◽

Peizhen Lv ◽

Huayu Wu ◽

Kun Zhang ◽

Fuben Xu ◽

...

Keyword(s):

Gene Expression ◽

Schwann Cells ◽

Neurotrophic Factors ◽

Underlying Mechanism ◽

Pharmacological Properties ◽

Neuroprotective Effects ◽

Antioxidative Effects ◽

Underlying Mechanisms ◽

And Function

Derived fromRhodiola roseaL., which is a popular plant in Eastern Europe and Asia, salidroside has pharmacological properties including antiviral, anticancer, hepatoprotective, antidiabetic, and antioxidative effects. Recent studies show that salidroside has neurotrophic and neuroprotective effects. However, the effect of salidroside on Schwann cells (SCs) and the underlying mechanisms of the salidroside-induced neurotrophin secretion have seldom been studied. In this study, the effect of salidroside on the survival, proliferation, and gene expression of Schwann cells lineage (RSC96) was studied through the examinations of the cell viability, proliferation, morphology, and expression of neurotrophic factor related genes including BDNF, GDNF, and CDNF at 2, 4, and 6 days, respectively. These results showed that salidroside significantly enhanced survival and proliferation of SCs. The underlying mechanism might involve that salidroside affected SCs growth through the modulation of several neurotrophic factors including BDNF, GDNF, and CDNF. As for the concentration, 0.4 mM, 0.2 mM, and 0.1 mM of salidroside were recommended, especially 0.2 mM. This investigation indicates that salidroside is capable of enhancing SCs survival and function in vitro, which highlights the possibility that salidroside as a drug agent to promote nerve regeneration in cellular nerve scaffold through salidroside-induced neurotrophin secretion in SCs.

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(Pro)renin receptor: another member of the system controlled by angiotensin II?

Journal of the Renin-Angiotensin-Aldosterone System ◽

10.1177/1470320311423280 ◽

2011 ◽

Vol 13 (1) ◽

pp. 1-10 ◽

Cited By ~ 5

Author(s):

Luciana G Pereira ◽

Carine P Arnoni ◽

Edgar Maquigussa ◽

Priscila C Cristovam ◽

Juliana Dreyfuss ◽

...

Keyword(s):

Gene Expression ◽

Angiotensin Ii ◽

Mesangial Cells ◽

At1 Receptor ◽

Receptor Internalization ◽

Prorenin Receptor ◽

Receptor Blockers ◽

And Function ◽

At1 Receptor Blockers

The prorenin receptor [(P)RR] is upregulated in the diabetic kidney and has been implicated in the high glucose (HG)-induced overproduction of profibrotic molecules by mesangial cells (MCs), which is mediated by ERK1/2 phosphorylation. The regulation of (P)RR gene transcription and the mechanisms by which HG increases (P)RR gene expression are not fully understood. Because intracellular levels of angiotensin II (AngII) are increased in MCs stimulated with HG, we used this in vitro system to evaluate the possible role of AngII in (P)RR gene expression and function by comparing the effects of AT1 receptor blockers (losartan or candesartan) and (P)RR mRNA silencing (siRNA) in human MCs (HMCs) stimulated with HG. HG induced an increase in (P)RR and fibronectin expression and in ERK1/2 phosphorylation. These effects were completely reversed by (P)RR siRNA and losartan but not by candesartan (an angiotensin receptor blocker that, in contrast to losartan, blocks AT1 receptor internalization). These results suggest that (P)RR gene activity may be controlled by intracellular AngII and that HG-induced ERK1/2 phosphorylation and fibronectin overproduction are primarily induced by (P)RR activation. This relationship between AngII and (P)RR may constitute an additional pathway of MC dysfunction in response to HG stimulation.

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Simulated microgravity using the Random Positioning Machine inhibits differentiation and alters gene expression profiles of 2T3 preosteoblasts

AJP Cell Physiology ◽

10.1152/ajpcell.00222.2004 ◽

2005 ◽

Vol 288 (6) ◽

pp. C1211-C1221 ◽

Cited By ~ 94

Author(s):

Steven J. Pardo ◽

Mamta J. Patel ◽

Michelle C. Sykes ◽

Manu O. Platt ◽

Nolan L. Boyd ◽

...

Keyword(s):

Gene Expression ◽

Alkaline Phosphatase ◽

Bone Loss ◽

Simulated Microgravity ◽

Expression Profiles ◽

Gene Expression Profiles ◽

Underlying Mechanism ◽

Bone Biology ◽

Random Positioning Machine

Exposure to microgravity causes bone loss in humans, and the underlying mechanism is thought to be at least partially due to a decrease in bone formation by osteoblasts. In the present study, we examined the hypothesis that microgravity changes osteoblast gene expression profiles, resulting in bone loss. For this study, we developed an in vitro system that simulates microgravity using the Random Positioning Machine (RPM) to study the effects of microgravity on 2T3 preosteoblast cells grown in gas-permeable culture disks. Exposure of 2T3 cells to simulated microgravity using the RPM for up to 9 days significantly inhibited alkaline phosphatase activity, recapitulating a bone loss response that occurs in real microgravity conditions without altering cell proliferation and shape. Next, we performed DNA microarray analysis to determine the gene expression profile of 2T3 cells exposed to 3 days of simulated microgravity. Among 10,000 genes examined using the microarray, 88 were downregulated and 52 were upregulated significantly more than twofold using simulated microgravity compared with the static 1-g condition. We then verified the microarray data for some of the genes relevant in bone biology using real-time PCR assays and immunoblotting. We confirmed that microgravity downregulated levels of alkaline phosphatase, runt-related transcription factor 2, osteomodulin, and parathyroid hormone receptor 1 mRNA; upregulated cathepsin K mRNA; and did not significantly affect bone morphogenic protein 4 and cystatin C protein levels. The identification of gravisensitive genes provides useful insight that may lead to further hypotheses regarding their roles in not only microgravity-induced bone loss but also the general patient population with similar pathological conditions, such as osteoporosis.

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The GAGA factor is required in the early Drosophila embryo not only for transcriptional regulation but also for nuclear division

Development ◽

10.1242/dev.122.4.1113 ◽

1996 ◽

Vol 122 (4) ◽

pp. 1113-1124 ◽

Cited By ~ 1

Author(s):

K.M. Bhat ◽

G. Farkas ◽

F. Karch ◽

H. Gyurkovics ◽

J. Gausz ◽

...

Keyword(s):

Gene Expression ◽

Chromatin Remodeling ◽

In Vitro Transcription ◽

Drosophila Embryo ◽

Gaga Factor ◽

Early Drosophila Embryo ◽

Hypersensitive Sites ◽

And Function ◽

Stimulatory Factor

The GAGA protein of Drosophila was first identified as a stimulatory factor in in vitro transcription assays using the engrailed and Ultrabithorax promoters. Subsequent studies have suggested that the GAGA factor promotes transcription by blocking the repressive effects of histones; moreover, it has been shown to function in chromatin remodeling, acting together with other factors in the formation of nuclease hypersensitive sites in vitro. The GAGA factor is encoded by the Trithorax-like locus and in the studies reported here we have used the maternal effect allele Trl13C to examine the functions of the protein during embryogenesis. We find that GAGA is required for the proper expression of a variety of developmental loci that contain GAGA binding sites in their upstream regulatory regions. The observed disruptions in gene expression are consistent with those expected for a factor involved in chromatin remodeling. In addition to facilitating gene expression, the GAGA factor appears to have a more global role in chromosome structure and function. This is suggested by the spectrum of nuclear cleavage cycle defects observed in Trl13C embryos. These defects include asynchrony in the cleavage cycles, failure in chromosome condensation, abnormal chromosome segregation and chromosome fragmentation. These defects are likely to be related to the association of the GAGA protein with heterochromatic satellite sequences which is observed throughout the cell cycle.

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Dynamic changes in chromatin accessibility, altered adipogenic gene expression, and total versus de novo fatty acid synthesis in subcutaneous adipose stem cells of normal-weight polycystic ovary syndrome (PCOS) women during adipogenesis: evidence of cellular programming

Clinical Epigenetics ◽

10.1186/s13148-020-00970-x ◽

2020 ◽

Vol 12 (1) ◽

Author(s):

Karen L. Leung ◽

Smriti Sanchita ◽

Catherine T. Pham ◽

Brett A. Davis ◽

Mariam Okhovat ◽

...

Keyword(s):

Gene Expression ◽

Fatty Acid ◽

De Novo ◽

Polycystic Ovary ◽

Total Content ◽

Chromatin Accessibility ◽

Normal Weight ◽

Triacylglycerol Synthesis ◽

And Function

Abstract Background Normal-weight polycystic ovary syndrome (PCOS) women exhibit adipose resistance in vivo accompanied by enhanced subcutaneous (SC) abdominal adipose stem cell (ASC) development to adipocytes with accelerated lipid accumulation per cell in vitro. The present study examines chromatin accessibility, RNA expression and fatty acid (FA) synthesis during SC abdominal ASC differentiation into adipocytes in vitro of normal-weight PCOS versus age- and body mass index-matched normoandrogenic ovulatory (control) women to study epigenetic/genetic characteristics as well as functional alterations of PCOS and control ASCs during adipogenesis. Results SC abdominal ASCs from PCOS women versus controls exhibited dynamic chromatin accessibility during adipogenesis, from significantly less chromatin accessibility at day 0 to greater chromatin accessibility by day 12, with enrichment of binding motifs for transcription factors (TFs) of the AP-1 subfamily at days 0, 3, and 12. In PCOS versus control cells, expression of genes governing adipocyte differentiation (PPARγ, CEBPα, AGPAT2) and function (ADIPOQ, FABP4, LPL, PLIN1, SLC2A4) was increased two–sixfold at days 3, 7, and 12, while that involving Wnt signaling (FZD1, SFRP1, and WNT10B) was decreased. Differential gene expression in PCOS cells at these time points involved triacylglycerol synthesis, lipid oxidation, free fatty acid beta-oxidation, and oxidative phosphorylation of the TCA cycle, with TGFB1 as a significant upstream regulator. There was a broad correspondence between increased chromatin accessibility and increased RNA expression of those 12 genes involved in adipocyte differentiation and function, Wnt signaling, as well as genes involved in the triacylglycerol synthesis functional group at day 12 of adipogenesis. Total content and de novo synthesis of myristic (C14:0), palmitic (C16:0), palmitoleic (C16:1), and oleic (C18:1) acid increased from day 7 to day 12 in all cells, with total content and de novo synthesis of FAs significantly greater in PCOS than controls cells at day 12. Conclusions In normal-weight PCOS women, dynamic chromatin remodeling of SC abdominal ASCs during adipogenesis may enhance adipogenic gene expression as a programmed mechanism to promote greater fat storage.

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Helicases FANCJ, RTEL1 and BLM Act on Guanine Quadruplex DNA in Vivo

Genes ◽

10.3390/genes10110870 ◽

2019 ◽

Vol 10 (11) ◽

pp. 870 ◽

Cited By ~ 4

Author(s):

Peter Lansdorp ◽

Niek van Wietmarschen

Keyword(s):

Gene Expression ◽

Genome Stability ◽

Cell Function ◽

Historical Context ◽

Telomere Maintenance ◽

Dna Structures ◽

G4 Dna ◽

Guanine Quadruplex

Guanine quadruplex (G4) structures are among the most stable secondary DNA structures that can form in vitro, and evidence for their existence in vivo has been steadily accumulating. Originally described mainly for their deleterious effects on genome stability, more recent research has focused on (potential) functions of G4 structures in telomere maintenance, gene expression, and other cellular processes. The combined research on G4 structures has revealed that properly regulating G4 DNA structures in cells is important to prevent genome instability and disruption of normal cell function. In this short review we provide some background and historical context of our work resulting in the identification of FANCJ, RTEL1 and BLM as helicases that act on G4 structures in vivo. Taken together these studies highlight important roles of different G4 DNA structures and specific G4 helicases at selected genomic locations and telomeres in regulating gene expression and maintaining genome stability.

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Gene Expression in Granulosa Cells From Small Antral Follicles From Women With or Without Polycystic Ovaries

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/jc.2019-00780 ◽

2019 ◽

Vol 104 (12) ◽

pp. 6182-6192 ◽

Cited By ~ 9

Author(s):

Lisa Ann Owens ◽

Stine Gry Kristensen ◽

Avi Lerner ◽

Georgios Christopoulos ◽

Stuart Lavery ◽

...

Keyword(s):

Gene Expression ◽

Granulosa Cells ◽

Polycystic Ovary ◽

Ovarian Tissue ◽

Polycystic Ovaries ◽

Vitro Fertilization ◽

Antral Follicles ◽

Oocyte Aspiration ◽

And Function

Abstract Context Polycystic ovary syndrome (PCOS) is the most common cause of anovulation. A key feature of PCOS is arrest of follicles at the small- to medium-sized antral stage. Objective and Design To provide further insight into the mechanism of follicle arrest in PCOS, we profiled (i) gonadotropin receptors; (ii) characteristics of aberrant steroidogenesis; and (iii) expression of anti-Müllerian hormone (AMH) and its receptor in granulosa cells (GCs) from unstimulated, human small antral follicles (hSAFs) and from granulosa lutein cells (GLCs). Setting GCs from hSAFs were collected at the time of cryopreservation of ovarian tissue for fertility preservation and GLCs collected during oocyte aspiration before in vitro fertilization/intracytoplasmic sperm injection. Participants We collected hSAF GCs from 31 women (98 follicles): 10 with polycystic ovaries (PCO) and 21 without. GLCs were collected from 6 women with PCOS and 6 controls undergoing IVF. Main Outcome Measures Expression of the following genes: LHCGR, FSHR, AR, INSR, HSD3B2, CYP11A1, CYP19, STAR, AMH, AMHR2, FST, INHBA, INHBB in GCs and GLCs were compared between women with PCO and controls. Results GCs in hSAFs from women with PCO showed higher expression of LHCGR in a subset (20%) of follicles. Expression of FSHR (P < 0.05), AR (P < 0.05), and CYP11A1 (P < 0.05) was lower, and expression of CYP19A1 (P < 0.05), STAR (P < 0.05), HSD3B2 (P = NS), and INHBA (P < 0.05) was higher in PCO GCs. Gene expression in GL cells differed between women with and without PCOS but also differed from that in GCs. Conclusions Follicle arrest in PCO is characterized in GCs by differential regulation of key genes involved in follicle growth and function.

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P0658MATRIX AND FLOW MODULATE GENE EXPRESSION IN A 3D VASCULARISED TUBULE MODEL

Nephrology Dialysis Transplantation ◽

10.1093/ndt/gfaa142.p0658 ◽

2020 ◽

Vol 35 (Supplement_3) ◽

Author(s):

Julie Williams ◽

Sanlin Robinson ◽

Babak Alaei ◽

Kimberly Homan ◽

Maryam Clausen ◽

...

Keyword(s):

Gene Expression ◽

Extracellular Matrix ◽

Impact Analysis ◽

Cell Types ◽

Drug Uptake ◽

Shear Stresses ◽

The Matrix ◽

And Function

Abstract Background and Aims Questions abound regarding the translation of in vitro 2D cell culture systems to the human setting. This is especially true of the kidney in which there is a complex hierarchical structure and a multitude of cell types. While it is well accepted that extracellular matrix plays a large part in directing cellular physiology emerging research has highlighted the importance of shear stresses and flow rates too. To fully recapitulate the normal gene expression and function of a particular renal cell type how important is it to completely reconstitute their in vivo surroundings? Method To answer this question, we have cultured proximal tubular (PT) epithelial cells in a 3-dimensional channel embedded within an engineered extracellular matrix (ECM) under physiological flow that is colocalised with an adjacent channel lined with renal microvascular endothelial cells that mimic a peritubular capillary. Modifications to the system were made to allow up to 12 chips to be run in parallel in an easily handleable form. After a period of maturation under continuous flow, both cell types were harvested for RNAseq analyses. RNA expression data was compared with cells cultured under static 2-dimensional conditions on plastic or the engineered ECM. Additionally, the perfusion of glucose through this 3D vascularised PT model has been investigated in the presence and absence of known diabetes modulating agents. Results PCA of RNAseq data showed that a) static non-coated, b) static matrix-coated and c) flow matrix-coated conditions separated into 3 distinct groups, while cell co-culture had less impact. Analysis of transcriptomic signatures showed that many genes were modulated by the matrix with additional genes influenced under flow conditions. Several of these genes, classified as transporters, are of particular importance when using this model to assess drug uptake and safety implications. Co-culture regulated some interesting genes, but fewer than anticipated. Preliminary experiments are underway to monitor glucose uptake and transport between tubules under different conditions. Conclusion We have developed a medium throughput system in which matrix and flow modulate gene expression. This system can be used to study the physiology of molecular cross-talk between cells. Ongoing analysis will further consider relevance to human physiology.

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Evaluation of Lentiviral-Mediated Expression of Sodium Iodide Symporter in Anaplastic Thyroid Cancer and the Efficacy of In Vivo Imaging and Therapy

Journal of Oncology ◽

10.1155/2011/178967 ◽

2011 ◽

Vol 2011 ◽

pp. 1-7 ◽

Cited By ~ 5

Author(s):

Chien-Chih Ke ◽

Ya-Ju Hsieh ◽

Luen Hwu ◽

Fu-Hui Wang ◽

Fu-Du Chen ◽

...

Keyword(s):

Gene Expression ◽

Sodium Iodide ◽

Cell Function ◽

Tumor Imaging ◽

Promising Result ◽

Anaplastic Thyroid Cancer ◽

Anaplastic Thyroid Carcinoma ◽

Sodium Iodide Symporter

Anaplastic thyroid carcinoma (ATC) is one of the most deadly cancers. With intensive multimodalities of treatment, the survival remains low. ATC is not sensitive to131I therapy due to loss of sodium iodide symporter (NIS) gene expression. We have previously generated a stable human NIS-expressing ATC cell line, ARO, and the ability of iodide accumulation was restored. To make NIS-mediated gene therapy more applicable, this study aimed to establish a lentiviral system for transferring hNIS gene to cells and to evaluate the efficacy of in vitro and in vivo radioiodide accumulation for imaging and therapy. Lentivirus containing hNIS cDNA were produced to transduce ARO cells which do not concentrate iodide. Gene expression, cell function, radioiodide imaging and treatment were evaluated in vitro and in vivo. Results showed that the transduced cells were restored to express hNIS and accumulated higher amount of radioiodide than parental cells. Therapeutic dose of131I effectively inhibited the tumor growth derived from transduced cells as compared to saline-treated mice. Our results suggest that the lentiviral system efficiently transferred and expressed hNIS gene in ATC cells. The transduced cells showed a promising result of tumor imaging and therapy.

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