Studies of hTERT DNA methylation assays on the human age prediction

Ye, Xin; Dong, Kaikai; Cao, Fangqi; Tian, Yantao; Sun, Jing; Liu, Wenbin; Shi, Ping

doi:10.1007/s00414-019-02076-3

Cited by 7 publications

(6 citation statements)

References 21 publications

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“…In our cohort, we observed high TERT mRNA expression levels in tumors with pTERT WT, which could be explained by the presence of other alterations such as hypermethylation, chromosomal rearrangements, ampli cations or transcriptional fusions 15,16 . These alterations may not be activated by environmental factors in cell lines or not manifest in young patients, such as pTERT methylation which has been shown to increase with age 23 . Interestingly, if we strati ed patients according to the type of pTERT mutation, we found a signi cant association between the C250T mutation and higher TERT mRNA expression levels compared to the C228T mutation; unlike previous studies in which they observed otherwise 36,37 .…”

Section: Discussionmentioning

confidence: 99%

“…DNA from normal skin tissue was used as the baseline for pTERT methylation. Methylation levels of pTERT were measured according to the protocol described by Xin Y et al 23 . PCR products were analyzed in a 3% agarose gel (Figure S2) and the intensity of the ampli ed bands was measured by densitometry using the ImageJ program (version 1.53e for Windows, NIH, Bethesda, Maryland).…”

Section: Methylation Speci C Pcr (Ms-pcr)mentioning

confidence: 99%

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pTERT C250T mutation: a potential biomarker of poor prognosis in metastatic melanoma

García

Domínguez

Martínez-Illescas

et al. 2022

Preprint

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Background Melanoma is the most aggressive form of skin cancer and the leading cause of death from cutaneous tumors. Several studies have associated alterations in the TERT promoter region (pTERT) with gene overexpression, aggressiveness and poor prognosis of the disease. The aim of this study was to clarify the role of pTERT molecular status in paired samples of primary melanoma and metastasis using tissue and plasma to establish a correlation with disease progression and survival. Methods A total of 88 FFPE tissue samples from 53 patients with advanced melanoma were analyzed. Of these, 35 had paired samples. We also examined cfDNA samples from plasma of 25 patients. pTERT mutational status was analyzed using pyrosequencing and digital droplet PCR (ddPCR). pTERT methylation status was evaluated by methylation-specific PCR (MS-PCR), while TERT mRNA expression levels were measured using reverse transcription quantitative PCR (RT-qPCR). Results and discussion We detected a good correlation between primary tumors and metastases in pTERT mutation and methylation status. We were also able to detect pTERT mutations in plasma samples. Interestingly, the C250T mutation was associated with worse survival and higher TERT mRNA expression, compared to the other most common mutation: C228T. In addition, hyper-methylation of the promoter region seems to be related to the progression of pTERT WT patients. These results suggest that TERT gene alterations plays an important role during tumor progression, with the detection of the C250T mutation in tissue and plasma as a potential biomarker of poor prognosis in patients with advanced melanoma.

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Section: Discussionmentioning

confidence: 99%

Section: Methylation Speci C Pcr (Ms-pcr)mentioning

confidence: 99%

pTERT C250T mutation: a potential biomarker of poor prognosis in metastatic melanoma

García

Domínguez

Martínez-Illescas

et al. 2022

Preprint

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“…S1 ). MS-PCR was carried out according to the protocol described by Xin Y et al [ 25 ] ( Table S3 ). Non-bisulfite-treated DNA, methylated DNA (Merck, Darmstadt, Germany), unmethylated DNA (Qiagen) and NTC were used in each run as controls.…”

Section: Methodsmentioning

confidence: 99%

pTERT C250T mutation: A potential biomarker of poor prognosis in metastatic melanoma

Blanco-García,

Ruano,

Blanco Martínez-Illescas

et al. 2023

Heliyon

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“…Several studies reported the hTERT expressions using immunological methods viz. real-time polymerase chain reaction (PCR), Western blotting, immunoprecipitation, peptide mass spectrometry, gel electrophoresis, immunofluorescence, and confocal microscopy [18,19], but these methods did not report the actual concentration of hTERT. The majority of studies use reverse transcription-PCR (RT-PCR) with a probe on cellular mRNA to detect hTERT, but the presence of telomerase-active mRNA accounts for some discrepancy in results [20,21].…”

Section: Introductionmentioning

confidence: 99%

Biofabricated functionalized graphene quantum dots (fGQDs): unraveling its fluorescence sensing mechanism of human telomerase reverse transcriptase (hTERT) antigen and in vitro bioimaging application

Tade¹,

Patil²

2022

Biomed. Mater.

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Lung cancer (LC) is a deadly malignancy that is posing a serious threat to human health. Therefore, early detection of LC biomarkers is the key to reducing LC-related fatalities. Herein, we present the first fluorescent-based selective detection of LC biomarker human telomerase reverse transcriptase (hTERT) using polyethyleneimine (PEI) functionalized graphene quantum dots (fGQDs). One-pot in situ synthesis of amine-functionalized GQDs was accomplished by hydrothermal carbonization of biowaste-derived cellulose and PEI. Synthesized fGQDs were characterized by various analytical techniques. Synthesized fGQDs not only exhibited enhanced fluorescence life-time but also excellent stability in the different solvents compared to bare GQDs. The surface activation of hTERT-Ab by carbodiimide chemistry (EDC-NHS) resulted in stacking interactions with fGQDs, involving adsorption-desorption as well as competitive mechanisms. The higher inherent affinity of hTERT-Ag (hTERT antigen) for hTERT-Ab (hTERT antibody) resulted in complex formation and recovery of fGQD fluorescence. As a result, this fluorescence sensing demonstrated a greater linear detection range (0.01 ng mL-1 to 100 µg mL-1) as well as a notable low detection limit (36.3 pg mL-1). Furthermore, the fabricated immunosensor (Ab@fGQDs) has excellent stability and performance in real samples, with an average recovery of 97.32%. The results of cytotoxicity and cellular bioimaging study in A549 cells show that fGQDs can be used for additional nanotherapeutics and biological applications.

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Studies of hTERT DNA methylation assays on the human age prediction

Cited by 7 publications

References 21 publications

pTERT C250T mutation: a potential biomarker of poor prognosis in metastatic melanoma

pTERT C250T mutation: a potential biomarker of poor prognosis in metastatic melanoma

pTERT C250T mutation: A potential biomarker of poor prognosis in metastatic melanoma

Biofabricated functionalized graphene quantum dots (fGQDs): unraveling its fluorescence sensing mechanism of human telomerase reverse transcriptase (hTERT) antigen and in vitro bioimaging application

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