References of "Turtoi, Andrei"
     in
Bookmark and Share    
Full Text
Peer Reviewed
See detailNovel comprehensive approach for accessible biomarker identification and absolute quantification from precious human tissues
Turtoi, Andrei ULg; Dumont, Bruno ULg; Greffe, Yannick et al

in Journal of Proteome Research (2011), 10(7), 3160-82

The identification of specific biomarkers obtained directly from human pathological lesions remains a major challenge, because the amount of tissue available is often very limited. We have developed a ... [more ▼]

The identification of specific biomarkers obtained directly from human pathological lesions remains a major challenge, because the amount of tissue available is often very limited. We have developed a novel, comprehensive, and efficient method permitting the identification and absolute quantification of potentially accessible proteins in such precious samples. This protein subclass comprises cell membrane associated and extracellular proteins, which are reachable by systemically deliverable substances and hence especially suitable for diagnosis and targeted therapy applications. To isolate such proteins, we exploited the ability of chemically modified biotin to label ex vivo accessible proteins and the fact that most of these proteins are glycosylated. This approach consists of three successive steps involving first the linkage of potentially accessible proteins to biotin molecules followed by their purification. The remaining proteins are then subjected to glycopeptide isolation. Finally, the analysis of the nonglycosylated peptides and their involvement in an in silico method increased the confident identification of glycoproteins. The value of the technique was demonstrated on human breast cancer tissue samples originating from 5 individuals. Altogether, the method delivered quantitative data on more than 400 potentially accessible proteins (per sample and replicate). In comparison to biotinylation or glycoprotein analysis alone, the sequential method significantly increased the number (≥30% and ≥50% respectively) of potentially therapeutically and diagnostically valuable proteins. The sequential method led to the identification of 93 differentially modulated proteins, among which several were not reported to be associated with the breast cancer. One of these novel potential biomarkers was CD276, a cell membrane-associated glycoprotein. The immunohistochemistry analysis showed that CD276 is significantly differentially expressed in a series of breast cancer lesions. Due to the fact that our technology is applicable to any type of tissue biopsy, it bears the ability to accelerate the discovery of new relevant biomarkers in a broad spectrum of pathologies. [less ▲]

Detailed reference viewed: 53 (25 ULg)
Full Text
Peer Reviewed
See detailInnovative Proteomics for the Discovery of Systemically Accessible Cancer Biomarkers Suitable for Imaging and Targeted Therapies
Turtoi, Andrei ULg; De Pauw, Edwin ULg; Castronovo, Vincenzo ULg

in American Journal of Pathology (2011), 178(1), 12-18

The discovery of biomarkers that are readily accessible through the circulating blood and are selectively overexpressed in pathological tissues has become a major research objective, particularly in the ... [more ▼]

The discovery of biomarkers that are readily accessible through the circulating blood and are selectively overexpressed in pathological tissues has become a major research objective, particularly in the field of oncology. Indisputably, this group of molecules has a high potential to serve as an innovative tool for effective imaging and targeted cancer therapy approaches. In this attractive therapeutic concept, specific cancer proteins are reached by intravenously administered ligands that are coupled to cytotoxic drugs. Such compounds are able to induce cancer destruction while sparing normal tissues. Owing to the performance of mass spectrometry technology, current high-throughput proteomic analysis allows for the identification of a high number of proteins that are differentially expressed in the cancerous tissues. However, such approaches provide no information regarding the effective accessibility of the biomarkers and, therefore, the possibility for these discovered proteins to be targeted. To bypass this major limitation, which clearly slows the discovery of such biomarkers, innovative methodological strategies have been developed to enrich the clinical specimens before the mass spectrometry analysis. The focus is laid on the group of proteins that are necessarily located either at the exterior face of the plasma membrane or in the extracellular matrix. The present review addresses the current technologies meant for the discovery and analysis of accessible antigens from clinically relevant samples. [less ▲]

Detailed reference viewed: 39 (14 ULg)
Full Text
Peer Reviewed
See detailProteomic and genomic modulations induced by gamma-irradiation of human blood lymphocytes.
Turtoi, Andrei ULg; Sharan, Rajesh; Srivastava, Alok et al

in International Journal of Radiation Biology (2010), 86(10), 888-904

PURPOSE: Quantitative evaluation of early response proteins (ERPRO) and early response genes (ERG) following γ-irradiation of human lymphocytes; identification of specific proteins and genes as candidate ... [more ▼]

PURPOSE: Quantitative evaluation of early response proteins (ERPRO) and early response genes (ERG) following γ-irradiation of human lymphocytes; identification of specific proteins and genes as candidate biomarkers for the development of a novel biodosimeter. MATERIALS AND METHODS: Human peripheral blood lymphocytes were exposed to clinically relevant doses (1, 2 and 4 Gy) of γ-radiation ex-vivo. Analyses of protein and gene expression modulation were conducted 2 h post-irradiation. Global modulations were monitored using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and DNA microarray analyses of the samples originating from one human donor. On the proteome level, both phosphorylated and non-phosphorylated proteins were considered. Proteins and genes of specific interest were further targeted using Western blot (WB) and real-time quantitative polymerase chain reaction (RT-qPCR) techniques, employing samples from several human donors (n=3). RESULTS: A set of ERPRO and ERG showing significant alterations 2 h post-γ-irradiation have been identified in human lymphocytes. The most radiation responsive genes and proteins indicated alterations of cellular structure (ß-actin, talin-1 [TLN1], talin-2, zyxin-2), immune and defence reactions (major histocompatibility complex binding protein-2 [MBP2], interleukin-17E and interferon-γ), cell cycle control (cyclin-dependent kinase inhibitor-1A [CDKN1A], mouse double minute-2, annexin-A6 [ANXA6], growth arrest and DNA-damage-inducible protein-α [GADD45A], proliferating cell nuclear antigen [PCNA], dual specificity phosphatase-2 and 8 [DUSP8]) as well as detoxification processes (peroxin-1) and apoptosis (B-cell lymphoma-2 binding component-3 [BBC3]). SUMMARY: The estimations of protein concentration modulation of TLN1 and CDKN1A, phosphorylation status of ANXA6 (dose range 0-2 Gy) and MBP2 as well as the alterations in the level of gene expressions of BBC3, DUSP8, GADD45A and PCNA appears to be of potential value for future biodosimetric applications. [less ▲]

Detailed reference viewed: 31 (3 ULg)
Full Text
Peer Reviewed
See detailNovel post-digest isotope coded protein labeling method for phospho- and glycoproteome analysis
Fleron, Maximilien ULg; Greffe, Yannick ULg; Musmeci, Davide ULg et al

in Journal of Proteomics (2010), 73(10), 1986-2005

In the field of proteomics there is an apparent lack of reliable methodology for quantification of posttranslational modifications. Present study offers a novel post-digest ICPL quantification strategy ... [more ▼]

In the field of proteomics there is an apparent lack of reliable methodology for quantification of posttranslational modifications. Present study offers a novel post-digest ICPL quantification strategy directed towards characterization of phosphorylated and glycosylated proteins. The value of the method is demonstrated based on the comparison of two prostate related metastatic cell lines originating from two distinct metastasis sites (PC3 and LNCaP). The method consists of protein digestion, ICPL labeling, mixing of the samples, PTM enrichment and MS-analysis. Phosphorylated peptides were isolated using TiO(2), whereas the enrichment of glycosylated peptides was performed using hydrazide based chemistry. Isolated PTM peptides were analyzed along with non enriched sample using 2D-(SCX-RP)-Nano-HPLC-MS/MS instrumentation. Taken together the novel ICPL labeling method offered a significant improvement of the number of identified (∼600 individual proteins) and quantified proteins (>95%) in comparison to the classical ICPL method. The results were validated using alternative protein quantification strategies as well as label-free MS quantification method. On the biological level, the comparison of PC3 and LNCaP cells has shown specific modulation of proteins implicated in the fundamental process related to metastasis dissemination. Finally, a preliminary study involving clinically relevant autopsy cases reiterated the potential biological value of the discovered proteins. [less ▲]

Detailed reference viewed: 48 (8 ULg)
Full Text
Peer Reviewed
See detailGene expression profile of human lymphocytes exposed to (211)At alpha particles
Turtoi, Andrei ULg; Brown, Ian; Schläger, Martin et al

in Radiation Research (2010), 174

In this study, the Whole Human Genome 44K DNA microarray assay was used for the first time to obtain gene expression profiles in human peripheral blood lymphocytes 2 h after exposure (in suspension) to 6 ... [more ▼]

In this study, the Whole Human Genome 44K DNA microarray assay was used for the first time to obtain gene expression profiles in human peripheral blood lymphocytes 2 h after exposure (in suspension) to 6.78 MeV mean energy alpha particles from extracellular (211)At. Lymphocytes were exposed to fluences of 0.3-9.6 x 10(6) alpha particles/cm(2) [corresponding to mean absorbed alpha-particle doses (D(alpha)) of 0.05-1.60 Gy] over 30 min. Significantly modulated expression was identified in 338 early-response genes. Up-regulated expression was evident in 183 early-response genes, while the remaining 155 were down-regulated. Over half of the up-regulated genes and 40% of the down-regulated genes had a known biological process related primarily to cell growth and maintenance and cell communication. Genes associated with cell death were found only in the up-regulated genes and those with development only in the down-regulated genes. Eight selected early-response genes that displayed a sustained up- or down-regulation (CD36, HSPA2, MS4A6A, NFIL3, IL1F9, IRX5, RASL11B and SULT1B1) were further validated in alpha-particle-irradiated lymphocytes of two human individuals using the TaqMan(R) RT-qPCR technique. The results confirmed the observed microarray gene expression patterns. The expression modulation profiles of IL1F9, IRX5, RASL11B and SULT1B1 genes demonstrated similar trends in the two individuals studied. However, no significant linear correlation between increasing relative gene expression and the alpha-particle dose was evident. The results suggest the possibility that a panel of genes that react to alpha-particle radiation does exist and that they merit further study in a greater number of individuals to determine their possible value regarding alpha-particle biodosimetry. [less ▲]

Detailed reference viewed: 21 (2 ULg)
Full Text
See detailNovel Relative ICPL Based Quantitative Phospho- and Glycoproteome Analysis Method
Fleron, Maximilien ULg; Greffe, Yannick ULg; Massart, Anne-Cécile ULg et al

Poster (2010, April 16)

Large scale proteomic analysis remains challenging partially because proteins are inhomogeneous and often influenced by a variety of structural modifications. In particular, these specific chemical ... [more ▼]

Large scale proteomic analysis remains challenging partially because proteins are inhomogeneous and often influenced by a variety of structural modifications. In particular, these specific chemical modifications called posttranslational modifications (PTM) are crucial determinants for the protein function and biological role. Up to now there have been a growing number of studies describing the enrichment and identification of PTM. However, a significant dearth of data offering a reliable methodology for PTM quantification does exist. The present work aims at developing a label based protein PTM quantification strategy and demonstrating its value on comparative analysis of cells originating from two distinct prostate metastasis sites. PC3 and LNCaP cells isolated from bone and lymph node prostate cancer metastasis sites respectively, were lysed and spiked with three non-human proteins serving as internal standards. Following this, the samples were reduced and alkylated, digested with trypsin and subjected to peptide ICPL (isotope coded protein label) labeling. The two peptide containing samples were joined together followed by the affinity isolation of phospho- (using TiO2 metal affinity chromatography) and glycopeptides (oxidized glycans were bound on hydrazide resin). The enriched fraction as well as the flow-through were analyzed on a 2D-(SCX and C18-RP)-nano-HPLC system. The peptide identification and quantification was conducted using electrospray ion-trap mass spectrometer (Bruker, HCT-ultra). Validation of the differentially modulated proteins was conducted in several biological and technical replicates using the label free MSe based quantification strategy. This PTM based, novel relative protein quantification using post-digest ICPL has detected over 598 individual proteins. Of these more than 95 % have been successfully quantified. PTM enrichment methodologies allowed an isolation rate of 91 % and 50 % for phosphorylated and glycosylated proteins respectively. The detailed comparison of PC3 and LNCaP cells has shown specific overexpression of selected proteins indicating differences between these two prostate metastatic cell lines. Several of these modulated proteins have been previously described to be related to prostate cancer (e.g. annexin A2 and vimentin) while others could be considered as potentially novel. These proteins might be implicated in the fundamental process related to metastasis dissemination. However, because of the known discrepancy between cell systems and clinical material, the present study can be regarded only as a step towards elucidation of these complex interactions. [less ▲]

Detailed reference viewed: 24 (4 ULg)
Full Text
Peer Reviewed
See detailProteomic analysis of human pancreas cancers for the identification of targetable biomarkers
Castronovo, Vincenzo ULg; Wang, Ying Hong; Musmeci, Davide et al

(2010)

Detailed reference viewed: 20 (2 ULg)
Full Text
Peer Reviewed
See detailIsotope coded protein label quantification of serum proteins--comparison with the label-free LC-MS and validation using the MRM approach.
Turtoi, Andrei ULg; Mazzucchelli, Gabriel ULg; De Pauw, Edwin ULg

in Talanta (2010), 80(4), 1487-95

Protein quantification based upon mass spectrometry is gaining ground in diverse applications of biological and clinical relevance. The present article focuses on one of the most complex biological fluids ... [more ▼]

Protein quantification based upon mass spectrometry is gaining ground in diverse applications of biological and clinical relevance. The present article focuses on one of the most complex biological fluids - serum - and provides a novel ICPL based quantification protocol. The results are compared to a label-free (data independent alternate scanning) absolute quantification method. The validation is performed using MRM based protein quantification technique. Regarding the ICPL approach, serum samples used in this study were depleted of high abundant proteins, labeled with ICPL and fractionated according to their respective pI (3-5, 5-7 and 7-12). The samples were further subjected to tryptic digestion followed by treatment with the Glu-C enzyme. The peptides were analyzed on a 2D-nano-LC system using four different concentrations of salt injections (45, 75, 150 and 500 mM ammonium acetate). The LC system was connected on-line with the electrospray ion-trap mass spectrometer. For the label-free quantification the serum samples were depleted and digested with trypsin. A proteome-wide comparison was performed using highly reproducible LC and data independent alternate scanning in conjunction with a high mass accuracy orthogonal time-of-flight mass spectrometer. Selected proteins, found by both methods, were validated using the MRM approach. For this purpose non-depleted tryptically digested serum samples were analyzed by LC coupled with a triple-quadrupole MS. The relative protein quantification using ICPL and mass spectrometry allowed for the detection of approximately 200 proteins, whereas about 2/3 of those contained the ICPL label and could therefore be quantified. Label-free approach used no fractionation, less sample and was able to identify and quantify over 110 proteins. The identified proteins covered generally 3-4 orders of magnitude of protein concentration in human serum. Changes in relative abundance of eight proteins were validated using MRM. This study, for the first time, shows the ability of the relative protein quantification based upon ICPL and 2D-LC-MS/MS to quantify serum biomarkers. It provides two additional label-free approaches that could validate and bring additional value to the label-based results, offering a starting point for comprehensive proteomics studies aiming at revealing biomarkers of clinical relevance. [less ▲]

Detailed reference viewed: 194 (49 ULg)
Full Text
Peer Reviewed
See detailVersican overexpression in human breast cancer lesions: Known and new isoforms for stromal tumor targeting.
Kischel, Philippe ULg; Waltregny, David ULg; Dumont, Bruno ULg et al

in International Journal of Cancer = Journal International du Cancer (2010), 126(3), 640-50

Proteoglycans play a key role in cancer development and progression by participating in the constitution of a specific fertile tumor microenvironment. As they are largely overexpressed in the malignant ... [more ▼]

Proteoglycans play a key role in cancer development and progression by participating in the constitution of a specific fertile tumor microenvironment. As they are largely overexpressed in the malignant stroma, proteoglycans provide a reservoir of potential new targets for anticancer therapies, because they can serve to convey toxic payloads in the close proximity of cancer cells and subsequently destroy them. In this context, versican, a proteoglycan largely overexpressed in several solid cancers, bears the potential to be such an ideal target. As 4 main versican isoforms have been characterized, we sought to determine which isoform could represent the best target in human breast cancer. We used a series of 10 primary breast cancer lesions that were characterized as overexpressing the versican protein, when compared with matched normal breast tissues, using shotgun mass spectrometry and immunohistochemistry experiments. Quantitative polymerase chain reaction and western-blotting experiments were used to evaluate versican isoform expression in breast cancer/normal tissue pairs for which ARN quality was excellent. All known isoforms were significantly overexpressed in the malignant lesions, both at the mRNA and at the protein levels. In the course of this study, we also identified and cloned a new alternatively spliced versican isoform, referred to as V4, which was also found to be upregulated in human breast cancer. This study provides for the first time a comprehensive mRNA and protein analysis of versican isoforms expression in human breast tissues, and offers insights into which therapeutic strategy would be best suited to target versican in human breast cancer lesions. [less ▲]

Detailed reference viewed: 146 (51 ULg)
Full Text
See detailSecretion and maturation of toxins in the venom duct of Conustextile
Dobson, Rowan ULg; Corbesier, Corine; Collodoro, Mike et al

Conference (2009, December 02)

Detailed reference viewed: 19 (6 ULg)
See detailLe ciblage therapeutique: vers une guerre propre et efficace contre le cancer
Castronovo, Vincenzo ULg; Waltregny, David ULg; Detry, Olivier ULg et al

Scientific conference (2009, October)

Detailed reference viewed: 124 (15 ULg)
Full Text
Peer Reviewed
See detailRelative serum protein quantification based upon ICPL and 2D-LC-MS identifies potential frailty biomarkers in elderly patients
Turtoi, Andrei ULg; Mazzucchelli, Gabriel ULg; Dobson, Rowan ULg et al

Poster (2009, June 01)

This study shows the ability of the ICPL and nano-HPLC-MS/MS to perform relative quantification and identification of serum protein biomarkers. Frailty is a geriatric syndrome that is commonly associated ... [more ▼]

This study shows the ability of the ICPL and nano-HPLC-MS/MS to perform relative quantification and identification of serum protein biomarkers. Frailty is a geriatric syndrome that is commonly associated with the decline in multisystemic reserve, cognition and sensory capabilities. It is negatively influencing the outcome of a disease prolonging the patient’s recovery. The discrepancy between the actual and the biological age brings the uncertainty of predicting frailty in a given individual. This study is addressing the problem of finding suitable biomarkers that bear the ability to objectively predict frailty in elderly patients. It furthermore provides a robust method for reliable relative quantification of serumproteins. Serum samples used in this study were divided into six groups regarding the patient’s disease (hip-fracture, infection and cardiac decompensation) and frailty status (frail or robust). The individual sera were pooled and a volume of 20 µL was depleted of high abundantproteins. After labeling with ICPL (isotope coded protein label), serum proteins were fractionated according to their respective pI (0-3, 4-7 and 8-12). The samples were further subjected to tryptic digestion followed by the treatment with the Glu-C enzyme. The peptides were analyzed on the 2D-nano-HPLC system (Ulimate 3000®) using four different concentrations of salt injections (45, 75, 150 and 500 mM ammonium acetate). The HPLC system was connected on-line with the electrospray ion-trap mass spectrometer Esquire HCT ultra®. The relative protein quantification using ICPL and mass spectrometry allowed for comparison of six patient groups with respect to a standard sample. The latter represented a group of healthy old subjects. This technique allowed for the detection of approx. 200 proteins, whereas about 50 % of those contained the ICPL label and could therefore be quantified. The identified proteins covered 3 – 4 orders of magnitude ofprotein concentration in human serum. Several proteins displayed a significant modulation allowing for some preliminary conclusions to be drawn. At this point it can be stated that significantly elevated levels of C-reactive protein (factor 12) and alpha-1-antichimotrypsin (f. 4) proved to be potentially good indicators of frailty. Increased concentrations of alpha-1-microglobulin (f. 4) and alpha-2HS-glycoprotein (f. 2) have been found in the robust patients, whereas no significant concentration alteration could be detected in the frail groups. These results refer to the acute phase response since the samples were collected immediately after patient hospitalization. Current investigations addressing the later sampling times should shed more light on the suitability of these markers to predict frailty in elderly patients. At this stage it is obvious that although several markers are found to be in common for all the frail or robust patients, the disease status additionally complicates the biomarker signature. Therefore a more individualized approach should also be considered, where depending on the age and clinical findings a more defined group of markers should be selected to address the problem of frailty. [less ▲]

Detailed reference viewed: 51 (5 ULg)
Full Text
Peer Reviewed
See detailLe ciblage thérapeutique : vers une guerre propre et efficace contre le cancer
Castronovo, Vincenzo ULg; Waltregny, David ULg; Detry, Olivier ULg et al

in Revue Médicale de Liège (2009), 64

One promising avenue towards the development of more selective, better anticancer drugs consists in the targeted delivery of bioactive compounds to the tumor environment by means of binding molecules ... [more ▼]

One promising avenue towards the development of more selective, better anticancer drugs consists in the targeted delivery of bioactive compounds to the tumor environment by means of binding molecules specific for tumor-associated biomarkers. Eligibility of such markers for therapeutic use implies ideally three criteria : (i) accessibility from the bloodstream, (ii) expression at sufficient level and (iii) no (or much lower) expression in normal tissues. Most current discovery strategies (such as biomarker searching into body fluids) provide no clue as to whether proteins of interest are accessible, in human tissues, to suitable high-affinity ligands, such as systemically delivered monoclonal antibodies. Innovative proteomic technologies are able to identify such accessible biomarkers and represent a key step in the clinical development of such target therapies. [less ▲]

Detailed reference viewed: 142 (35 ULg)
Full Text
Peer Reviewed
See detailEffect of 211At alpha-particle irradiation on expression of selected radiation responsive genes in human lymphocytes
Turtoi, Andrei ULg; Schneeweiss, Frank H.A.

in International Journal of Radiation Biology (2009), 85(5), 403-12

Detailed reference viewed: 19 (8 ULg)
Full Text
See detailUntersuchung frühzeitiger Reaktionen lymphozytärer Proteine auf γ-Bestrahlung humanen Vollbluts und deren Dosisabhängigkeit – Voraussetzung für die Entwicklung eines individuellen strahlenbiologischen Dosimeters
Turtoi, Andrei ULg

Book published by Forschungszentrum Jülich (2008)

The present thesis is concerned with the issues involved in obtaining reliable experimental data permitting a retrospective assessment of radiation-induced doses at the time of application or ... [more ▼]

The present thesis is concerned with the issues involved in obtaining reliable experimental data permitting a retrospective assessment of radiation-induced doses at the time of application or contamination. In order to provide prompt medical treatment of those injured in accidents with ionizing radiation, biological procedures that can be implemented swiftly and at an early stage are required both to determine the radiation dose originally received as well as to assess the course of the dose-dependent biological reactions on the basis of individual sensitivity to radiation. To this end, in the present thesis the lymphocyte proteins (phosphoproteins and total proteins) in blood taken from test subjects who had been exposed to γ-radiation (applied dose: 0-4 Gy) were analysed just 15 minutes after completing irradiation by means of 2D gel electrophoresis. Only those early-response proteins (ERPROs) that displayed a significant radiation-induced change were identified by nano-HPLC-MS/MS. For validation purposes, the dose-dependent gene expression of some of these proteins was determined by RT-qPCR. The following ERPROs displayed pronounced early reactions in the form of changes of concentration in comparison to unirradiated control samples: talin-1, talin-2, β-actin, mutant β-actin, peroxin-1 and also the phosphoproteins annexin-A6, MHC-binding protein-2, zyxin-2, interleukin-17E and phosphoglycerate kinase-1. The majority of the lymphocyte ERPROs represent proteins responsible for changes to the cytoskeleton, proliferation and cell cycle, modulation of immunoreactions as well as protein degradation and energy production. Other cellular processes may not have been determined due to the sensitivity restrictions of the 2D-PAGE and MS methods, but cannot be excluded. Gene expression studies revealed that a combination of methods, comprising RT-qPCR and 2D-PAGE as well as DNA microarray and Western blot, may in future be able to overcome these restrictions. The slopes of the curves from concentration measurements of various early response proteins after doses had been applied in the 0 and 4 Gy range yielded a characteristic arrangement or pattern representative of an individual. In case of contamination, this pattern prepared ex vivo serves as a reference for identifying the originally unknown dose, thus creating the necessary condition for applying an individual radiation biodosimeter. This thus provides for the first time an experimental means of biologically quantifying in retrospect radioactive doses in the 0 and 4 Gy range after a relatively short time. [less ▲]

Detailed reference viewed: 26 (1 ULg)
Full Text
Peer Reviewed
See detailEarly gene expression in human lymphocytes after gamma-irradiation-a genetic pattern with potential for biodosimetry.
Turtoi, Andrei ULg; Brown, Ian; Oskamp, Dominik et al

in International Journal of Radiation Biology (2008), 84(5), 375-87

PURPOSE: Identification of early radiation response genes (ERG) in human lymphocytes after gamma-irradiation by using the whole-human-genome DNA-microarrays and the evaluation of their possible role in ... [more ▼]

PURPOSE: Identification of early radiation response genes (ERG) in human lymphocytes after gamma-irradiation by using the whole-human-genome DNA-microarrays and the evaluation of their possible role in rapid radiation biodosimetry by applying real-time quantitative polymerase chain reaction (RT-qPCR) methodology for validation in a small group of human individuals. MATERIALS AND METHODS: Whole blood from a healthy human donor was exposed at 37 degrees C to 137Cs gamma-radiations (absorbed dose: 1-4 Gy). Fifteen minutes following irradiation the lymphocytes were isolated from the blood (for 2 h at 20 degrees C) and their gene expression was investigated using the DNA-microarrays. Subsequently, 14 genes were selected and validated using the TaqMan probes based upon the RT-qPCR assay within a group of 6 human donors. RESULTS: A dose-related relative change in quantitative gene expression using the DNA-microarray assay was demonstrated in 24 of 102 genes. Up-regulation of expression was observed in 15 genes: CD69 (CD69 molecule), CDKN1A (cyclin-dependent kinase inhibitor 1A), EGR1 (early growth response 1), EGR4 (early growth response 4), FLJ35725 (chromosome 4 ORF 23), hCG2041177 (hCG - human Celera Genome), hCG1643466.2, IFN-gamma (interferon-gamma), ISG20L (interferon stimulated exonuclease gene 20 kDa - like 1), c-JUN (jun oncogene), MDM2 (mouse double minute 2), MUC5B (mucine), PLK2 (polo-like kinase 2), RND1 (rho-family GTPase 1) and TNFSF9 (tumour necrosis factor superfamily member 9). Down-regulation of expression was found in the remaining nine genes: GRIK3 (glutamate receptor ionotropic kainate 3), hCG1985174, hCG1998530, hCG2038519, OCLN (occludin), RPL10A (ribosomal protein L10a), SERHL2 (serine hydrolase-like 2), SGK3 (serum/glucocorticoid regulated kinase 3) and STARD13 (START domain containing 13). CONCLUSION: A significant correlation between absorbed radiation dose and change in relative gene expression was particularly evident for EGR1, EGR4, IFN-gamma, c-JUN and TNFSF9 (p < or = 0.05). Results warrant the further investigation of these ERG as potential biodosimetric markers. [less ▲]

Detailed reference viewed: 40 (5 ULg)
Full Text
Peer Reviewed
See detailEarly response of lymphocyte proteins after gamma-radiation
Turtoi, Andrei ULg; Srivastava, Alok; Sharan, Rajesh et al

in Journal of Radioanalytical & Nuclear Chemistry (2007), 274(2), 435-39

Detailed reference viewed: 17 (0 ULg)