Genomic Instability in Patients with Non-Small Cell Lung Cancer Assessed by the Arbitrarily Primed Polymerase Chain Reaction

Markovic, Jasna; Stojšić, Jelena; Zunic, Svetlana; Ruždijić, Sabera; Tanić, Nikola

doi:10.1080/07357900701708385

Cited by 20 publications

(19 citation statements)

References 32 publications

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“…The needle was dipped in the PCR master mix for 2 min and then discarded. The PCR products were reamplified with the same primers used for AP-PCR reactions at high-stringency conditions specific for each particular primer [5]. The reamplified material was administrated on 1.5% agarose gels, purified using DNA Extraction Kit (Fermentas Life Sciences, Lithuania) and cloned with GeneJet TM PCR Cloning Kit (Fermentas Life Sciences, Lithuania) according to manufacturers' instructions.…”

Section: Isolation Cloning and Sequencing Of Variant Electrophoreticmentioning

confidence: 99%

“…Five primers (p53 A, MDR A, E5A p53, GAPDH A, and GAPDH S) produced informative sequence alterations differentiating normal tissue from tumor tissue. Primer sequences (p53 A, MDR A, E5A p53 and GAPDH A), AP-PCR conditions and reaction mixtures are as described previously [5] including one additional primer GAPDHS: 5 -CGG AGT CAA CGG ATT TGG TCG TAT -3 ; AP-PCR low-stringency conditions: 95 • C 30 ; 50 • C 2 ; 72 • C 1 ; AP-PCR high-stringency conditions: 95 • C 30 ; 70 • C 1 ; 72 • C 1 and AP-PCR reaction mixture: 0.4 mM each dNTP; 2.5 mM MgCl 2 ; 5 M of a primer; 1 U of Taq DNA polymerase.…”

Section: Ap-pcr Dna Fingerprintingmentioning

confidence: 99%

“…Following our previous study of genomic instability in NSCLC tissue samples [5] we aimed to identify genes that alter during NSCLC promotion and progression. For the past 10 years this efficient screening method has been used to detect novel genetic alterations in various cancers, including pancreatic and colorectal carcinomas [6,7] lung cancer [8,9], breast cancer [10], hepatocellular carcinoma [11] and cholangiocarcinoma [12].…”

Section: Introductionmentioning

confidence: 99%

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Identification of genes associated with non-small-cell lung cancer promotion and progression

et al. 2010

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Section: Isolation Cloning and Sequencing Of Variant Electrophoreticmentioning

confidence: 99%

Section: Ap-pcr Dna Fingerprintingmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Identification of genes associated with non-small-cell lung cancer promotion and progression

et al. 2010

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“…The high incidence of genomic instability in lung cancers is wellestablished and in some cases it has been associated with poor prognosis (1)(2)(3). To guarantee genomic stability under the conditions of continuous genotoxic stress, a coordinated DNA damage surveillance system and several DNA repair pathways have been evolutionally developed.…”

Section: Introductionmentioning

confidence: 99%

Non–Small Cell Lung Cancer Exhibits Transcript Overexpression of Genes Associated with Homologous Recombination and DNA Replication Pathways

et al. 2009

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Genes involved in DNA repair and replication have been recently investigated as predictive markers of response to chemotherapy in non-small cell lung cancer (NSCLC). However, few data on the expression of these genes in tumor compared with corresponding normal lung are available. The aim of this study was to evaluate differential mRNA levels of 22 DNA repair genes of five different DNA repair pathways: direct, base excision, nucleotide excision (NER), double-strand break (DSBR), and postreplicative repair. In addition, six genes involved in DNA replication (REP) and three telomere maintenance genes were investigated. Total RNAs extracted from fresh-frozen tumors and corresponding normal tissues of 50 consecutive chemo-naïve resected NSCLC patients were analyzed. Transcript levels were quantified by real-time PCR. A significant overexpression was detected in 20 of 30 (67%) genes, mostly belonging to DSBR pathways, whereas others (XPA, XPC, and UBE2N; 10%) were significantly underexpressed. For 7 of 30 (23%) genes, mostly belonging to NER pathway, no significant difference between paired tumor and normal samples was observed. Transcript overexpression of DSBR and REP genes was significantly higher in poorly differentiated carcinomas and DSBR levels were higher in men compared with women. The transcriptional overexpression of four genes (XRCC5, TOP3B, TYMS, and UNG) showed significant correlation with a shorter patients' outcome at the univariate, whereas only stage of disease appeared as an independent factor affecting prognosis, as assessed by multivariate analysis. In conclusion, genes belonging to DNA repair/replication pathways are overexpressed in NSCLC and are associated with a more aggressive phenotype. [Cancer Res 2009;69(8):3390-6]

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“…Studies conducted on the lung cancer have shown frequent genomic imbalances (Markovic et al, 2008), and it is suggested that chromosomal instability plays an important role in its progression (Ninomiya et al, 2006). Structural chromosomal rearrangements and losses/gains from non-disjunctions and multipolar mitoses are thought to result in aneuploidy.…”

Section: Discussionmentioning

confidence: 99%

Inflammatory response to isocyanates and onset of genomic instability in cultured human lung fibroblasts

Mishra¹,

Bhargava²,

Raghuram³

et al. 2009

Genet. Mol. Res.

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ABSTRAcT. Lungs comprise the primary organ exposed to environmental toxic chemicals, resulting in diverse respiratory ailments and other disorders, including carcinogenesis. Carcinogenesis is a multi-stage phenomenon, which involves a series of genetic alterations that begin with genomic instability provoked by certain factors such as inflammation and DNA damage and end with the development of cancer. Isocyanates such as methyl isocyanate are the chief metabolic intermediates in many industrial settings with diverse applications; exposure to them can lead to severe hypersensitive, mutagenic and genotoxic alterations. We examined the molecular mechanisms underlying isocyanate-mediated inflammatory responses and their probable role in the onset of genomic instability in cultured IMR-90 human lung fibroblasts. The isocyanates induced inflammation, resulting in extensive DNA damage, evidenced by increases in ATM, ATR, γH2AX, and p53 expression levels. The apoptotic index also increased. Chromosomal anomalies in treated cells included over-expression of centrosome protein and variable amplification of inter-simple sequence repeats, further demonstrating isocyanate-induced genomic insta- bility. This information could be useful in the design of new approaches for risk assessment of potential industrial disasters.

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Genomic Instability in Patients with Non-Small Cell Lung Cancer Assessed by the Arbitrarily Primed Polymerase Chain Reaction

Cited by 20 publications

References 32 publications

Identification of genes associated with non-small-cell lung cancer promotion and progression

Identification of genes associated with non-small-cell lung cancer promotion and progression

Non–Small Cell Lung Cancer Exhibits Transcript Overexpression of Genes Associated with Homologous Recombination and DNA Replication Pathways

Inflammatory response to isocyanates and onset of genomic instability in cultured human lung fibroblasts

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