Highly sensitive detection of telomerase activity in tumor cells by cascade isothermal signal amplification based on three-way junction and base-stacking hybridization

Zhao, Yongxi; Lin, Qi; Chen, Feng; Zhao, Yue; Fan, Chunhai

doi:10.1016/j.bios.2012.10.009

Cited by 59 publications

(24 citation statements)

References 42 publications

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“…Beyond this point further division induces damage in the coding regions of the chromosome and causes cell senescence. Almost all human cancers and immortalized cell lines have highly active telomerase (40,41). The E7 protein of HPV-16 is a small, nuclear polypeptide of 100 amino acids.…”

Section: Hpv Virologymentioning

confidence: 99%

Relevance of infection with human papillomavirus: The role of the p53 tumor suppressor protein and E6/E7 zinc finger proteins

Ruttkay-Nedecký

Jiménez

Nejdl

et al. 2013

International Journal of Oncology

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Human papillomaviruses (HPV) are small circular, double-stranded DNA viruses infecting epithelial tissues. HPV types can be classified both as high-risk or low-risk. Of the more than 120 different identified types of HPV, the majority are involved in infections of the genital tract, cancer of the cervix, vulva, vagina and penis, and of non-anogenital localizations, such as the head and neck areas. From the point of view of the infection, human papillomaviruses have developed several molecular mechanisms to enable infected cells to suppress apoptosis. This review provides a comprehensive and critical summary of the current literature that focuses on cervical carcinoma and cancer of the head and neck caused by HPV. In particular, we discuss HPV virology, the molecular mechanisms of carcinogenesis, the role of the tumor suppressor protein p53 and the E6/E7 zinc finger proteins. Classification of HPV according to diagnosis is also described.

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Section: Hpv Virologymentioning

confidence: 99%

Relevance of infection with human papillomavirus: The role of the p53 tumor suppressor protein and E6/E7 zinc finger proteins

Ruttkay-Nedecký

Jiménez

Nejdl

et al. 2013

International Journal of Oncology

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“…Accordingly,t elomerase is regarded as ac ommon cancer marker for early diagnosis and prognosis.W eu sed telomerase substrate (TS) primert op erform telomerase-catalyzed extension reactions. [42][43][44][45] As mall number of TS primers are extended with 3' telomericr epeatsi nt he presence of telomerase (Scheme 1B). These extension products can act as templates for DNA amplification by zero-background HDA.…”

Section: Resultsmentioning

confidence: 99%

Zero‐Background Helicase‐Dependent Amplification and Its Application to Reliable Assay of Telomerase Activity in Cancer Cell by Eliminating Primer–Dimer Artifacts

Chen

Zhang

et al. 2016

ChemBioChem

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Primer-dimer artifacts resulting from unintended template-independent primer-primer interactions often hinder the specific amplification of nucleic acids. We demonstrate, for the first time, zero-background helicase-dependent amplification (HDA), with low concentrations of both ATP and dNTPs. This strategy achieved the reliable evaluation of telomerase activity in cancer cells by eliminating primer-dimer artifacts, which have plagued many previous methods with reduced specificity. We found that the performance of the telomerase assay by zero-background HDA was negatively affected by highly concentrated cellular proteins. This inhibitory effect is attributed to the binding of DNA templates to proteins, thus making them unavailable for polymerases. However, gold nanoparticles were demonstrated to highly attenuate such inhibition by abundant proteins, and to enhance the assay sensitivity and reliability when the reaction was performed with concentrated cell extracts.

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“…Recently, some highly sensitive and ultrafast detection methods of telomerase activity by isothermal amplification have reported . PLLBA may also be applicable to isothermal amplification such as loop‐mediated isothermal amplification (LAMP).…”

Section: Discussionmentioning

confidence: 99%

Development of a novel telomerase assay using the PPDK–luciferin–luciferase detection system

2013

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Telomerase participates in malignant transformation or immortalization of cells, and has attracted attention as an anticancer drug screening and diagnostic tumor marker. We developed a novel telomerase assay called the PPDK-luciferin-luciferase system bioluminescence assay (PLLBA) using pyruvate phosphate dikinase (PPDK). In this assay, pyrophosphate produced by the telomerase reaction and polymerase chain reaction (PCR) is converted to ATP by PPDK, and ATP is detected by the firefly luciferin-luciferase reaction. In this work, telomerase substrate was obtained in accordance with the telomeric repeat amplification protocol (TRAP). Telomerase-positive (500 cells/assay), -inactive (heated for 10 min at 85 °C) and -negative (only Chaps lysis buffer) samples were used. As a result, the findings clearly showed that the signal-to-noise (S/N) ratio of the positive cells was 39.5. After the telomerase reaction and PCR, PLLBA was completed ~ 120 s later. A high level of reproducibility was obtained with - coefficient of variation (CV) of 4.1% (positive cells). The detection limit for cells using telomerase was one cell per assay. This assay for telomerase activity was also shown to be adaptable to human cancer-derived cell lines.

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Highly sensitive detection of telomerase activity in tumor cells by cascade isothermal signal amplification based on three-way junction and base-stacking hybridization

Cited by 59 publications

References 42 publications

Relevance of infection with human papillomavirus: The role of the p53 tumor suppressor protein and E6/E7 zinc finger proteins

Relevance of infection with human papillomavirus: The role of the p53 tumor suppressor protein and E6/E7 zinc finger proteins

Zero‐Background Helicase‐Dependent Amplification and Its Application to Reliable Assay of Telomerase Activity in Cancer Cell by Eliminating Primer–Dimer Artifacts

Development of a novel telomerase assay using the PPDK–luciferin–luciferase detection system

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