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Ponchel, Frédérique; Toomes, Carmel; Bransfield, Kieran; Leong, Fong T; Douglas, Susan; Field, Sarah; Bell, Sandra; Combaret, Valérie; Puisieux, Alain; Mighell, Alan J.; Robinson, Philip A.; Inglehearn, Chris F.; Isaacs, John D.; Markham, Alex F.

doi:10.1186/1472-6750-3-18

Cited by 288 publications

(106 citation statements)

References 27 publications

(34 reference statements)

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“…This phenomena is similar to Mamedova, et al's report, the antenna effect may happen between Taq enzyme and CdTe quantum dots, may result in a significant increase of mQDs emission [29][30].…”

Section: Effects Of Taq Enzyme On Pl Property Of Mqdssupporting

confidence: 74%

Real time PCR based on Fluorescent Quenching of Mercaptoacetic Acid-Modified CdTe Quantum Dots for Ultrasensitive Specific Detection of Nucleic Acids

Cui¹,

Li²,

Huang³

et al. 2010

Nano BioMed ENG

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A ultra-sensitive, highly specific, real-time polymerase chain reaction system based on mercaptoacetic acid-modified CdTe nanocrystals(mQDs) is reported. With the addition of 3 nm mQDs into the PCR reagent, the photoluminescent(PL) intensities of mQDs decreased gradually as the DNA templates and PCR cycles increased, in an approximate negative linear relation to the DNA concentration logarithm or cycles, the PL peaks exhibited red-shifts synchronously. Mg 2+ ions decreased the PL intensity of mQDs in a dose-dependent means, and Taq DNA polymerase enhanced the PL intensity of mQDs in a dose-dependent means. Real-time PCR based on mQDs showed an increased sensitivity at least 103 fold higher than that based on SYBR Green I. The specificity of PCR was enhanced in the PCR reagent with less than 1.33mg/mL mQDs. The potential mechanism is also discussed. This novel PCR system based on mQDs has great potential in applications such as ultra-sensitive specific DNA or RNA detection, dynamic molecular imaging, and photoelectric biosensors.

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Section: Effects Of Taq Enzyme On Pl Property Of Mqdssupporting

confidence: 74%

Real time PCR based on Fluorescent Quenching of Mercaptoacetic Acid-Modified CdTe Quantum Dots for Ultrasensitive Specific Detection of Nucleic Acids

Cui¹,

Li²,

Huang³

et al. 2010

Nano BioMed ENG

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“…A real-time PCR (SYBR-Green-I based fluorescence, Bio-Rad, Hercules, CA) assay was developed where the threshold cycle number (C t ) is proportional to the logarithm of the initial amount of template used in the reaction (molecules per reaction). Thus, the number of molecules in an unknown sample can be calculated by comparing with a standard curve generated from samples in which the amount of DNA is known (20). First, a RT-PCR primer set 1 (B-actin forward, 5Ј-GCCTTTTATGGTAATAATGAGAG; and GFP reverse, 5Ј-GTGAACAGCTCCTCGCCCTTGC) was designed to amplify a 296-bp fragment that detects the nonrecombined B-actinlox-EGFP-lox-kRASG12D, whereas primer set 2 (B-actin forward, 5Ј-GAAGTTGACTCCAGATGGTCAC; and RAS reverse, 5Ј-CTACGCCATCAGCTCCAACTAC) amplifies a 193-bp fragment found only after recombination (SI Fig.…”

Section: Discussionmentioning

confidence: 99%

“…Thus, this line was used in all subsequent analysis. To evaluate the efficiency and dynamics of the heat shockinducible Cre/Lox recombination approach, a real-time PCR-based assay (20) was designed to access recombination efficiency in line 25A at the genomic DNA level ( Fig. 1J and SI Fig.…”

Section: Human Krasg12d Expression Can Be Induced In Transgenic Zebramentioning

confidence: 99%

Heat shock-inducible Cre/Lox approaches to induce diverse types of tumors and hyperplasia in transgenic zebrafish

Langenau

Keefe

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

166

125

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RAS family members are among the most frequently mutated oncogenes in human cancers. Given the utility of zebrafish in both chemical and genetic screens, developing RAS-induced cancer models will make large-scale screens possible to understand further the molecular mechanisms underlying malignancy. We developed a heat shock-inducible Cre/Lox-mediated transgenic approach in which activated human kRASG12D can be conditionally induced within transgenic animals by heat shock treatment. Specifically, double transgenic fish Tg(B-actin-LoxP-EGFP-LoxP-kRASG12D; hsp70-Cre) developed four types of tumors and hyperplasia after heat shock of whole zebrafish embryos, including rhabdomyosarcoma, myeloproliferative disorder, intestinal hyperplasia, and malignant peripheral nerve sheath tumor. Using ex vivo heat shock and transplantation of whole kidney marrow cells from double transgenic animals, we were able to generate specifically kRASG12D-induced myeloproliferative disorder in recipient fish. This heat shock-inducible recombination approach allowed for the generation of multiple types of RAS-induced tumors and hyperplasia without characterizing tissue-specific promoters. Moreover, these tumors and hyperplasia closely resemble human diseases at both the morphologic and molecular levels.myeloproliferative disorder ͉ RAS ͉ rhabdomyosarcoma ͉ intestine ͉ malignant peripheral nerve sheath tumor R AS genes encode a family of 21-kDa proteins that switch between inactive GDP-bound (RAS-GDP) and active GTPbound (RAS-GTP) conformations. Once in its activated GTPbound form, RAS interacts with downstream effectors to modulate diverse cellular responses, including proliferation, differentiation, and survival (1). Point mutations within RAS family members often occur at codon 12, 13, or 61 (2), which abolish RAS-GTP hydrolysis and lead to constitutive activation of downstream signaling pathways. These activating mutations are common in human malignancies; for example, Ͼ90% of pancreatic adenocarcinomas, 50% of colorectal cancers, 25-50% of lung cancers, 5-35% of rhabdomyosarcomas (RMS), and 25-50% of myeloid leukemia have mutational activation of RAS family members. Among the three different human RAS genes (H-, N-,

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“…RT-PCR/Quantitative PCR-Transcript levels were measured by quantitative real-time PCR using previously described methods (22) using the following primer sequences: IL-6, 5Ј-CCTTCTCCACAAGCGCCTTC-3Ј (forward) and 5Ј-GGCA-AGTCTCCTCATTGAATC-3Ј (reverse); IL-8, 5Ј-ATGACTT-CCAAGCTGGCCGT-3Ј (forward) and 5Ј-CCTCTTCAAAA-ACTTCTCCACACC-3Ј (reverse); COX-2, 5Ј-TTCAAATGA-GATTGTGGAAAAAT-3Ј (forward) and 5Ј-AGATCATCTC-TGCCTGAGTATCTT-3Ј (reverse); ␤-actin, 5Ј-TCCGGAGA-CGGGGTCA-3Ј (forward) and 5Ј-CCTGCTTGCTGAT-CCA-3Ј (reverse). Quantitative PCRs were performed in triplicate using an iCycler (Bio-Rad).…”

Section: -(45-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4mentioning

confidence: 99%

Nuclear Factor-κB (NF-κB) Mediates a Protective Response in Cancer Cells Treated with Inhibitors of Fatty Acid Synthase

Lemmon¹,

Woo²,

Tully

et al. 2011

Journal of Biological Chemistry

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Overexpression of fatty acid synthase (FAS)4 is common in aggressive human cancers, and blocking FAS inhibits growth and leads to apoptosis in these cancer cells (1, 2). Although it is now well accepted that increased activity of this enzyme plays a key role in maintaining the metabolic stability of cancer cells, the molecular consequences of inhibiting FAS are still not well understood.One central molecular network commonly altered in cancer pathogenesis and also affected by cancer treatment is that regulated by the transcription factor NF-B. NF-B signaling is constitutively activated in many epithelial solid tumors and hematologic malignancies (3-5), and this activation appears to generally confer resistance to apoptosis as well as enhance growth properties of cancer cells. Consequently, numerous inhibitors of NF-B are being investigated for potential use in treating cancer (6, 7), and bortezomib, a proteosome inhibitor widely used for treating myeloma, is thought to act largely through inhibition NF-B by stabilization of the IB␣ subunit (8, 9). Interestingly, NF-B activity is also inhibited by a number of naturally occurring lipid compounds that have recognized anti-neoplastic properties, including curcumin, resveratol, and coix seed extract (10 -12), providing additional evidence to suggest that NF-B activity supports or promotes the malignant phenotype.NF-B activity does not uniformly contribute to malignancy, however, and in some situations, increased NF-B activity may actually suppress malignant characteristics of cells (13). For example, it has been shown that induction of p53 leads to activation of NF-B, correlating with the ability of p53 to induce apoptosis (14). Thus, at least in some cellular settings, inhibition or loss of NF-B activity abrogates p53-induced apoptosis, indicating that NF-B can be functional in p53-mediated cell death.The role of NF-B signaling in the response of cancer cells to chemotherapy also appears to depend on variables of the particular situation. In many circumstances, activation of NF-B by therapeutic agents appears to inhibit apoptosis and thus attenuates the response to these agents (15-17). However, activation of NF-B by cancer therapeutic agents appears to mediate cell death in other circumstances, including treatment with UV light (18), doxorubicin (19), and paclitaxel (20). In light of the general importance of NF-B to cellular physiology and response to stress and the expectation that manipulations of lipid metabolic pathways could affect NF-B signaling, we investigated the effects of inhibiting FAS on NF-B and the role of NF-B signaling in the response of lung cancer cells to this inhibition. 4 The abbreviations used are: FAS, fatty acid synthase; mIB␣, mutant IB␣; MTS, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium; PGE2, prostaglandin E2; PMA, phorbol 12-myristate 13-acetate; CE, cloning efficiency. EXPERIMENTAL PROCEDURES

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Untitled

Cited by 288 publications

References 27 publications

Real time PCR based on Fluorescent Quenching of Mercaptoacetic Acid-Modified CdTe Quantum Dots for Ultrasensitive Specific Detection of Nucleic Acids

Real time PCR based on Fluorescent Quenching of Mercaptoacetic Acid-Modified CdTe Quantum Dots for Ultrasensitive Specific Detection of Nucleic Acids

Heat shock-inducible Cre/Lox approaches to induce diverse types of tumors and hyperplasia in transgenic zebrafish

Nuclear Factor-κB (NF-κB) Mediates a Protective Response in Cancer Cells Treated with Inhibitors of Fatty Acid Synthase

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