Oxidative stress 2014

Sample & Assay Technologies Oxidative Stress and ROS Signaling

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� Overview� Oxidants & antioxidant in biological

systems

� Antioxidant defense mechanism

� Molecular targets of oxidative stress

� Detection of oxidative stress

� Research Solutions @ QIAGEN� Gene expression

� Epigenetics

� Functional studies

� Application Examples

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Sample & Assay Technologies Oxidative stress and ROS signaling

� Oxidative stress : imbalance between reactive oxygen species (ROS) and antioxidants.

� Under normal conditions, cells are able to balance the production of oxidants and antioxidants. Oxidative stress occurs when cells are subjected to excess levels of ROS or as a result of antioxidant depletion.

� ROS: chemically reactive molecules containing oxygen, highly reactive

� Under normal conditions, ROS are natural byproducts. It also can be generated by exogenous sources.

� Major sources of ROS: mitochondria, peroxisome, plasma membrane/cytoplasm.

Role of reactive oxygen species in cell signaling pathways, Biochemical and Biomedical Aspects of Oxidative Modification 2001

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Endogenous sources

MitochondriaPeroxisomes

Cytochrome P450

Exogenous sourcesUV light

Inflammatory cytokines

Pathogens

AntioxidantsVitamin ESeleniumLycopeneGreen tea

SNPsDNA repairOxidative enzymes

ROSOxidative Stress

Damage to DNA, RNA, protein and

lipids

Chromosomal instability, gene

mutations

Altered gene expression

Sample & Assay Technologies The biological importance of ROS

Physiological positive effects of ROS

� Kill microorganism

� Second messenger (H2O2)

� Cellular differentiation & proliferation

� Regulate signal transduction & transcription

� Cytokines and growth factor receptors (EGFR, PDGFR, VEGFR, receptors of TNFα, IFNγ and IL-1β)

� Non-receptor tyrosine kinases (JAK, Src, MAPK and PI3K)

� Protein tyrosine phosphatases

� Serine/Threonine kinases (Akt, PKC and MAPK)

� Nuclear transcription factors (NFκ B, AP-1, NFAT, HIF-1 and Nrf2)

Oxidative Stress and Oxidative Damage in Carcinogenesis, Toxicologic Pathology, 38: 96-109, 2010

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ROS

PLCγ

PI3KAkt ATM

NFkB

ERKJNKp38

NFAT

JAK

PKC

p53C-Jun

Nrf2

Gene expression changes

Sample & Assay Technologies Antioxidant defense mechanism

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� Inhibit excess ROS formation, capture radicals, maintain redox homeostasis

and correct mechanism of destroyed biomolecules

� Mediated through Antioxidant Responsive Element (ARE) sequence

� Controlled by nuclear transcription factors Nrf2, AP-1, NFκB, p53, NFAT

ROS Transcription factors

Transcription

ARE AREAntioxidant response elements (AREs)

Sample & Assay TechnologiesNrf2-mediated oxidative stress response

Nrf2: Nuclear factor-erythroid 2-related

factor is a basic leucine zipper (bZIP)

transcription factor that binds to ARE to

activate transcription.

Keap1: Kelch-like ECH-associated

protein 1. Keap1 interacts with Nrf2 in a

redox-sensitive manner.

Inactive Nrf2 is retained in the

cytoplasm by association with Keap1.

Upon exposure to oxidative stress, Nrf2

is phosphorylated in response to the

PKC, PI3K, and MAPK pathways. After

phosphorylation, Nrf2 is translocated to

the nucleus, binds AREs, and activates

detoxifying enzymes and antioxidant

enzyme.

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Sample & Assay Technologies Antioxidants – maintain oxidative balance

� Non-enzymatic antioxidants

� Alpha tocopherol (vitamin E)

� Ascorbic acid (vitamin C)

� Beta carotene

� Antioxidant enzymes

� Superoxide dismutase (SOD) O2−● + O2

−● + 2H+ � O2 + H2O2

� Catalase (CAT) 2 H2O2 � 2 H2O + O2

� Glutathione peroxidase (GPx) 2 G-SH + R-O-OH � G-S–S-G + R-OH + H2O

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Sample & Assay Technologies- 9 -

Antioxidant Action of Vitamin-C

Vitamin-C is a potent antioxidant and a regulator of redox-signal transduction

Under stress, Vitamin-C acts as a primary antioxidant in plasma and cells

Vitamin-C quenches ROS and serves as a cofactor of enzymes involved in the synthesis of collagen, carnitineAnd neurotransmitters

https://www.qiagen.com/geneglobe/pathwayview.aspx?pathwayID=34&

Sample & Assay Technologies Oxidative damage of ROS

� Aggregation and fragmentation

� Enzyme inhibition

Proteins� DNA strand breaks� Mutations leading

to cancer

DNA� Lipid peroxidation� Damage to

membranes and lipoproteins

Membrane

ROS

Oxidative stress, disease and aging

Review article: “Molecular targets of oxidative stre ss” Biochem. J. (2011) 434, 201

What is the cellular targets of ROS that account fo r the toxicity ?

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Sample & Assay Technologies Overview of oxidative stress & ROS

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Sample & Assay Technologies Diseases associated with oxidative stress

Diabetes

COPD

Neuro-degeneration

Asthma

Cardio-Vasculardisease

Dermatitis

Infertility

Athero-sclerosis

Cancer

OxidativeStress

inflammationChronic inflammation

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Oxidative Oxidative stress

Sample & Assay Technologies Oxidative stress links chronic inflammation & cancer

Oxidative stress, chronic inflammation, and cancer are closely linked

� Free radicals as a link of oxidative stress to inflammation and cancer

Oxidative stressOxidative stress

Carbonylic adducts Hydro & lipoperoxides Imbalance in DNA repair

Imbalance between pro & anti-apoptotic processes, altered gene expression & mutations

Chronic inflammation

Activation of Inflammatory cells

ROS production

LOXs, COXs, PHGPx iNOS

NFkBCytokines

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Sample & Assay Technologies ROS play multiple roles in the hallmarks of cancers

Review article: “Oxidative Stress, Tumor Microenvironment, and Metabolic Reprogramming: A Diabolic Liaison” International Journal of Cell Biology. 2012

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Sample & Assay Technologies

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� Oxidative stress is tightly linked to

mitochondrial dysfunction, and

mitochondria are both generators of

and targets of ROS

� In respond to cellular stress,

necrosis, apoptosis and autophagy

are activated.

� Mitochondrial turnover is dependent

on autophagy

� Dysregulated redox signaling or

mitochondrial dysfunction affect

autophagic activities

Mitochondria are the targets of ROS

Review article: “Autophagy, mitochondria and oxidative stress: cross-talk and redox signaling” Biochem. J. 2012, 441, 523

Sample & Assay Technologies miRNAs are modulators and targets of oxidative stress

“Inflammation and cancer: interweaving microRNA, free radical, cytokine and p53 pathways” Carcinogenesis, 31(1): 37–49, 2010

miRNAs are non-coding RNA molecules that modulate the stability and/or the translational efficiency of target messenger RNAs

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“Oxidative Stress and MicroRNAs in Vascular Diseases” Int. J. Mol. Sci. 2013, 14

Relevant miRNAs in oxidative stress response

ROS source

miRNAs

upregulated Tissue/organ Source Target Functions

H2O2 miR-200c

miR-141 endothelium

miR-200a myoblasts Human ZEB1 apoptosis

miR-200b

miR-429

H2O2 miR-200c ROS accumulation;

miR-141 miR-200a ovarian Human p38α improved response to

miR-200b adenocarcinomas chemotherapy

miR-429

H2O2 miR-200c primary Mouse Unknown Unknown

hippocampal

neurons

Obesity miR-200c miR-141 Heart Rat S6K1 compensatory/adaptive

mechanisms

Sample & Assay Technologies Detection of intracellular oxidative stress

� Direct approach� Detection of ROS / free radicals

� Colorimetric (ROS-dependent reactions), fluorometric (ROS-sensitive probes)

� Challenging: short-lived, highly reactive molecules

� Indirect approach� Antioxidant enzyme activity

� Detect oxidative stress damage products

� Measure cellular antioxidant capacity

� Simple: more stable modifications of macromolecules

� Wide range of techniques

Untreated Cells

Treated Cells

The cell-permeant dye H2-DCF-DA is non-fluorescent in a reduced state and exhibits fluorescence upon oxidation by ROS.

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Sample & Assay Technologies Detection of oxidative damage to macromolecules

� Lipid markers of oxidative stress� 4-Hydroxynonenal (HNE) is a by-product of lipid peroxidation, widely used as a

marker of chronic oxidative stress

� Protein markers of oxidative stress� Protein carbonylation – formation of carbonyl derivatives of Pro, Lys, Arg and Thr

residues – is the most common product of protein oxidation (ELISA assay).

� Advanced oxidation protein products (AOPP) are created when proteins react with chlorinated oxidants.

� DNA markers of oxidative stress� 8-hydroxydeoxyguanosine (8-OHdG)

� Antioxidant assays� Catalase activity, SOD activity

� Total antioxidant capacity

� GSH levels

Avery SV. Molecular targets of oxidative stress. Biochem J. 2011 Mar 1;434(2):201-10.

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� Overview� Oxidants & antioxidant in biological

systems

� Antioxidant defense mechanism

� Molecular targets of oxidative stress

� Detection of oxidative stress

� Research Solutions @ QIAGEN� Gene expression

� Epigenetics

� Functional studies

� Application Examples

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Sample & Assay Technologies Oxidative stress studies: experiment design

Functional studies

Epigenetics

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Gene expression

Sample & Assay Technologies Pathway-focused solution for gene expression analysis

RT2 Profiler PCR Arrays

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• 84 pathway-specific genes of interest

• 5 housekeeping genes

• Genomic DNA contamination control

• Reverse transcription controls (RTC) n=3

• Positive PCR controls (PPC) n=3

B2M, HPRT, RPL13A, GAPDH, HGDC

Sample & Assay Technologies Application 1: gene expression

In response to cellular oxidative stress, pathways including necrosis, apoptosis, and autophagy will be activated.

.How is cell fate determined under oxidative stress?

FASEB J. 2011 October; 25(10): 3366FASEB J. 2011 October; 25(10): 3366FASEB J. 2011 October; 25(10): 3366

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Experiment design

Treat with glucocorticoid

Isolate total RNA

Reverse transcription

(RT)

Data Analysis

cDNA synthesis

qPCR

Swiss-Webster mice MLO-Y4 cells

Application 1: gene expression

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1-2 pm EST, Feb. 3, 10, 17, 24, Monday, qPCR Technol ogy webinar series

https://www2.gotomeeting.com/register/311390954


Pathway-focused RT2 Profiler PCR Arrays

Apoptosis RT 2 Profiler PCR Array: 84 genes involved in programmed cell

death

Include: genes involved in induction of apoptosis, anti-apoptosis, regulation of

apoptosis, DEATH domain proteins, caspases and regulators

Autophagy RT 2 Profiler PCR Array: 84 genes involved in autophagy

Include: autophagy machinery components, regulation of autophagy

Oxidative stress RT 2 Profiler PCR Array: 84 genes involved in oxidative

stress


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human, mouse, rat, and up to 13 species


Oxidative Stress RT2 Profiler PCR Array

- human, mouse, rat, and more

Antioxidants:Glutathione Peroxidases (GPx): GPX1, GPX2, GPX3, GPX4, GPX5, GPX6, GPX7, GSTP1, GSTZ1.

Peroxiredoxins (TPx): PRDX1, PRDX2, PRDX3, PRDX4, PRDX5, PRDX6 (AOP2).

Other Peroxidases: CAT, CYBB, CYGB, DUOX1, DUOX2, EPX, LPO, MGST3, MPO, PTGS1, PTGS2 (COX2), PXDN, TPO, TTN.

Other Antioxidants: ALB, APOE, GSR, MT3, SELS, SOD1, SOD3, SRXN1, TXNRD1, TXNRD2.

Genes Involved in ROS Metabolism:Superoxide Dismutases (SOD): SOD1, SOD2, SOD3.

Other Genes Involved in Superoxide Metabolism: ALOX12, CCS, DUOX1, DUOX2, GTF2I, MT3, NCF1, NCF2, NOS2 (iNOS), NOX4, NOX5, PREX1, UCP2.

Other Genes Involved in ROS Metabolism: AOX1, BNIP3, EPHX2, MPV17, SFTPD.

Oxidative Stress Responsive Genes: APOE, ATOX1, CAT, CCL5 (RANTES), CYGB, DHCR24, DUOX1, DUOX2, DUSP1 (PTPN16), EPX, FOXM1, FTH1, GCLC, GCLM, GPX1, GPX2, GPX3, GPX4, GPX5, GPX6, GPX7, GSR, GSS, HMOX1, HSPA1A, KRT1, LPO, MBL2, MPO, MSRA, NQO1, NUDT1, OXR1, OXSR1, PDLIM1, PNKP, PRDX2, PRDX5, PRDX6 (AOP2), PRNP, RNF7, SCARA3, SELS, SEPP1, SIRT2, SOD1, SOD2, SQSTM1, SRXN1, STK25, TPO, TTN, TXN, TXNRD1, TXNRD2.

Oxygen Transporters: CYGB, MB.


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Results: Excess GC (glucocorticoid) decreased the expression of oxidative stress responsive genes


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Sample & Assay Technologies Application 1: oxidative stress & autophagy

Results:• Activation of the antioxidant genes positively correlated with activation of the autophagic

genes• No significant correlation between the activation of antioxidant and antiapoptosis genes

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Sample & Assay Technologies Application 1: gene expression in oxidative stress

RT2 Profiler PCR Arrays

� Apoptosis

� Autophagy

� Necrosis

� Cell Death Pathwayfinder

� Chemokines & Receptors

� Cytokines & Chemokines

� DNA Damage Signaling

� Hypoxia Signaling

� IL6/STAT3 Signalin

� Inflammatory Cytokines &

Receptors

� Inflammatory Response &

Autoimmunity

� Oxidative Stress

� JAK / STAT Signaling

� MAP Kinase Signaling

� mTOR Signaling

� NFkB Signaling

� TNF Signaling

� Toll-Like Receptor Signaling

� Transcription Factors

� p53 Signaling

� PI3K-AKT Signaling

� VEGF Signaling

� WNT Signaling

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Sample & Assay Technologies Oxidative stress: gene expression

Apoptosis:

Pro-Apoptotic: ABL1, APAF1, ATP6V1G2, BAX, BCL2L11, BIRC2 (c-IAP2), CASP1 (ICE), CASP3, CASP6,

CASP7, CASP9, CD40 (TNFRSF5), CD40LG (TNFSF5), CFLAR (CASPER), CYLD, DFFA, FAS (TNFRSF6),

FASLG (TNFSF6), GADD45A, NOL3, SPATA2, SYCP2, TNF, TNFRSF1A, TNFRSF10A (TRAIL-R), TP53.

Anti-Apoptotic: AKT1, BCL2, BCL2A1 (Bfl-1/A1), BCL2L1 (BCL-X), BIRC3 (c-IAP1), CASP2, IGF1R, MCL1,

TNFRSF11B, TRAF2, XIAP.

Autophagy: AKT1, APP, ATG12, ATG16L1, ATG3, ATG5, ATG7, BAX, BCL2, BCL2L1 (BCL-X), BECN1,

CASP3, CTSB, CTSS, ESR1 (ERa), FAS (TNFRSF6), GAA, HTT, IFNG, IGF1, INS, IRGM, MAP1LC3A,

MAPK8 (JNK1), NFKB1, PIK3C3 (VPS34), RPS6KB1, SNCA, SQSTM1, TNF, TP53, ULK1.

Necrosis: ATP6V1G2, BMF, C1orf159, CCDC103, COMMD4, CYLD, DEFB1, DENND4A, DPYSL4, EIF5B,

FOXI1, GALNT5, GRB2, HSPBAP1, JPH3, KCNIP1, MAG, OR10J3, PARP1 (ADPRT1), PARP2, PVR, RAB25,

S100A7A, SPATA2, SYCP2, TMEM57, TNFRSF1A, TXNL4B.

Cell Death PathwayFinder PCR Array

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Sample & Assay Technologies Applications of oxidative stress PCR Arrays

Publications Citing the Oxidative Stress PCR Array

• Susceptibility to Inhaled Flame-Generated Ultrafine Soot in Neonatal and Adult Rat Lungs, TOXICOLOGICAL SCIENCES 124(2), 472–486 (2011), Jackie K. W. Chan,* Michelle V, etc.

• Adaptive induction of NF-E2-related factor-2-driven antioxidant genes in endothelial cells in response to hyperglycemia, Am J Physiol Heart Circ Physiol 300: H1133–H1140, 2011, Zoltan Ungvari,1 Lora Bailey-Downs, etc.

• Tryptophan from human milk induces oxidative stress and upregulates the Nrf-2-mediated stress response in human intestinal cell lines. J Nutr. 2011 Aug;141(8):1417-23. Elisia I, Tsopmo A, Friel JK, etc.

• Age-related gene response of human corneal endothelium to oxidative stress and DNA damage. Invest Ophthalmol Vis Sci. 2011 Mar 1;52(3):1641-9. Joyce NC, Harris DL, Zhu CC

• Gene expression analysis of oxidative stress and apoptosis in proton-irradiated rat retina. In Vivo. 2010 Jul-Aug;24(4):425-30. Mao XW, Green LM, Mekonnen T, Lindsey N, Gridley DS

• Increased ANG II sensitivity following recovery from acute kidney injury: role of oxidant stress in skeletal muscle resistance arteries. Am J Physiol Regul Integr Comp Physiol. 2010 Jun;298(6):R1682-91. Phillips SA, etc.

• Effects of spaceflight on innate immune function and antioxidant gene expression. J Appl Physiol. 2009 Jun;106(6):1935-42. Baqai FP, Gridley DS, Slater JM, Luo-Owen X, Stodieck LS, Ferguson V, etc.

• Oxidative stress: molecular perception and transduction of signals triggering antioxidant gene defenses. Braz J Med BiolRes. 2005 Jul;38(7):995-1014. Scandalios JG

• Oxidative stress and gene regulation. Free Radic Biol Med. 2000 Feb 1;28(3):463-99. Allen RG, Tresini M

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Transcription factors Nrf2 and Nrf1 bind to the antioxidant response elements (ARE) and regulate genes involved in protecting cells from oxidative damages.

.How to measure the degree of antioxidant response?

Application 2: gene functions

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Sample & Assay Technologies Application 2: reporter assays for oxidative stress

Cignal Reporter Assays & Arrays

■ Dual-luciferase format

■ GFP format

■ DNA and Lenti assay

■ 45 signaling pathways

http://www.qiagen.com/search/cignal-reporter-assay-kits

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45-pathway array & 10-Pathway Arrays

Single Pathway Assay

Antioxidant Response ReporterCignal Finder Stress & Toxicity 10-pathway reporter Kit

Measure the transcriptional activity of the Nrf2 (NF-E2-related factor 2) and Nrf1 transcription factors. Nrf2 and Nrf1 bind to the antioxidant response elements (AREs) and regulate genes involved in protecting cells from oxidative damage.

Application 2: reporter assays for oxidative stress

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Answer two questions:

1. Whether hyperglycemia activates Nrf2 in endothelial cells?

2. Whether induction of Nrf2-regulated ROS detoxification systems protect

endothelial function in type 2 diabetes?

Experiment procedure: • Treat wild type and Nrf2 knockout mouse with a high-fat diet (HFD) and standard diet

• Use reporter assay to measure Nrf2 activity


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Published data using Cignal reporter assay kit


Results & conclusion: • Hyperglycemia induces oxidative stress

• Adaptive activation of the Nrf2/ARE pathway has a critical role in endothelial protection in response

to diabetic condition both in vitro and in vivo.


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Glucose (mmol/L)

Rel

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rf2

indu

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n (f

old

chan

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Glucose (mmol/L)

Rel

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e m

RN

A e

xpre

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old

chan

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Functional genomics:

�Assess RNA interference phenotypes

�Assess overexpression phenotypes

Functional proteomics:

�Analyze the effects of recombinant protein or peptide treatments

Drug discovery:

�Screen and validate small molecule drug candidates

Applications: Cignal reporter assay kits

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Download a white paper “Cignal Reporter Assay Kit: A high-performance tool for assessing the functions of genes, biologics, and small molecule compounds - (EN)”

http://www.qiagen.com/knowledge-and-support/resource-center/resource-download.aspx?id=6fc8144b-010b-43d8-9125-bf541a5d9c9c&lang=en

Benefits�Excellent sensitivity and specificity

�Maximal signal-to-noise ratio

�Minimal background transcription and rapid response

�Reduced experimental variation and greater reliability

Sample & Assay Technologies Application 3: antioxidant response mechanism

miRNAs play a regulatory role in Nrf2/Keap1 pathway

� How is Keap1 regulated?

� What is the mechanism of Nrf2-dependent antioxidant

response in cancer?

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• Dysregulation of the NRF2 signaling pathway is linked to cancer chemotherapy resistance

• Nrf2 may contribute to tumorigenesis• Nrf2 as a miRNA transcription factor• A number of miRNAs are regulated by Nrf2

Sample & Assay Technologies Application 3: miRNA expression profiling

Human breast cancer cell lines MDA-MB-231 and Hs578T, non-tumorigenic MCF-10A cells

� Profiled the expressions of 84 miRNAs using Human miFinder PCR Array (MIHS-001Z)

Experiment design:

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Sample & Assay Technologies miScript miRNA PCR Arrays

� 1 hour

� 2 minutes

� 2 hours

� 15 minutes

miFinder miRNA PCR Array

• Profiles the expression of the 84 most abundantly expressed and best characterized miRNAs in miRBase (www.miRBase.org);

• Maximize the likelihood of discovering miRNAs whose expression patterns correlate with the biological phenotypes;

• Available in Human, Mouse, Rat, Dog, Rhesus Macaque, more

miRNA PCR Array• miFinder• Serum & Plasma• Brain Cancer• Breast Cancer• Ovarian Cancer• Prostate Cancer • Cancer PathwayFinder• Apoptosis• Inflammatory Response and

Autoimmunity

Workflow

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Sample & Assay Technologies Application 3: miRNA expression profiling

� Conclusion: miR-200a regulates the Keap1/Nrf2 pathway in mammary epithelium ; Epigenetic therapy can restore the miR-200a regulation of Keap1 expression, therefore to reactivate Nrf2-dependent antioxidant pathway in breast cancer.

� Measured Nrf2 ARE promoter activity using luciferas e reporter assay

� Epigenetic therapy restored miR-200 expression and reduced Keap1 level

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• miR-200c is a tumor suppressor miRNA, involved in cancers such as breast, ovarian, endometrial, colorectal cancer, and pancreatic cancer

Normal cell: MCF-10A

Bre

ast c

ell l

ine:

MD

A-M

B-2

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Mature miRNAs Fold changes

has-miRNA-141 -869.07

has-miRNA-200c -422.65

has-miRNA-2302c -66.41

has-miRNA-196b -65.95

has-miRNA-143 -40.32

has-miRNA-122 -18.68

has-miRNA-7f -15.07

… …

Sample & Assay Technologies miRNA assays and reagents from QIAGEN

Target Identification Expression

Isolation

Function

miRNA mimics & inhibitors

miScript miRNA PCR Arrays

miRNeasy

miRNAStudies

Feb 6-27, Thursday : 4 part miRNA focused webinar series, monthlyhttps://www2.gotomeeting.com/register/152689794

RT² Profiler™ microRNA Targets PCR Arrays

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Sample & Assay Technologies Oxidative stress and DNA methylation

Stress & Toxicity PathwayFinder DNA Methylation PCR Array

Oxidative Stress: BNIP3, CYP1A1, GPX3, PRDX2, PTGS2 (COX2), SCARA3.

DNA Damage: ATM, BRCA1, CCND1, GADD45A, GADD45G, MLH1, MSH2, TP53, XPC.

Unfolded Protein Response: INSIG1, UBE2G2.

Hypoxia Signaling: CSTB, RARA.

Heat Shock Protein: DNAJC15.

Growth Arrest & Senescence: ATM, CDKN1A (p21CIP1/WAF1), GADD45A, GDF15 (PLAB), MSH2, TP53.

Inflammation: PRDX2, PTGS2 (COX2).

Anti-Apoptosis: BNIP3, CDKN1A (p21CIP1/WAF1), PRDX2.

Proliferation & Carcinogenesis: BRCA1, CCND1, CDKN1A (p21CIP1/WAF1), CYP1A1, PRDX2, PTGS2

(COX2), TP53.

EpiTect Methyl qPCRArray workflow

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Sample & Assay Technologies We provide service – send samples to us & receive results

. Whole Genome� Illumina Gene Expression Profiling� Illumina Genotyping

. Pathway / Focused Panel� Mutation Profiling� Methylation� PCR Array� miRNA PCR Array

. Individual Gene / Locus� Mutation Detection� Methylation� qPCR

. Sample Preparation – DNA, RNA Extraction and Purification� Cells, Tissue or Biofluids� Fixed Tissue� Small Sample

. http://www.qiagen.com/products/catalog/services/

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� Gene Expression� RT2 Profiler PCR Arrays & Assays� Custom PCR Arrays

� Epigenetics� miScript miRNA PCR Arrays & Assays� EpiTect Methyl qPCR Arrays & Assays� qBiomarker Somatic Mutation PCR Arrays� qBiomarker Copy Number PCR Arrays

� Functional Studies� Cignal Reporter Assays� SureSilencing shRNA Plasmid� Flexitube siRNA� Flexiplate siRNA, AllStar Control

Summary

QIAGEN offers many products for oxidative stress studies

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Sample & Assay Technologies Pathway-focused solutions @ QIAGEN

Complete systems: sample prep through data analysis

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http://www.qiagen.com/

http://www.qiagen.com/geneglobe/

Sample & Assay Technologies- 46 -

Thank You for Attending!

Are you ready to try?Contact Technical Support

North America:

• [email protected]• [email protected]

Phone: 1-800-362-7737

International customers:

[email protected]

Webinar related questions: [email protected]

Oxidative stress 2014

Technology

Transcript of Oxidative stress 2014