Increased Levels Of Telomerase Research Articles

The role of ceranib-2 and its nanoform on the decrease of telomerase levels in human non-small cell cancer.

Ceranib-2, an acid ceramidase (AC) inhibitor, can inhibit cancer cell proliferation and tumor development. However, poor water solubility and low cellular bioavailability limit its efficacy in cancer treatment. This study aimed to investigate the cell death induced by ceranib-2 and its solid lipid nanoformulation (ceranib-2-SLN) produced by the hot homogenization technique and the synergistic relationship between ceramide and telomerase in vitro and in silico. Furthermore, this study proved the possible mechanism of ceranib-2-induced AC inhibition by in silico studies. The effective cytotoxic concentrations of ceranib-2, telomerase level, and changes in ceramide levels were measured by MTT colorimetric cytotoxicity assay, ELISA, and LC/MS/MS methods, respectively. TEM results showed that ceranib-2-SLN was 13-fold smaller than the size of ceranib-2. Ceranib-2 and ceranib-2-SLN had IC50 concentrations of 31.62 (± 2.1) and 27.69 (± 1.75) µM in A549, and 48.79 (± 1.56) and 67.98 (± 2.33) in Beas-2B cells. These compounds simultaneously increased ceramide levels and decreased telomerase levels in A549 cells. Ceranib-2 increased telomerase levels while decreasing ceramide levels in Beas-2B cells. It was shown how the synergistic impact of ceranib-2-induced ceramide production and ceramide-induced telomerase level reduction on cytotoxicity in A549 cells. Ceranib-2-SLN was discovered to be more cytotoxic on cancer cells than ceranib-2, suggesting that it could be a promising option for the development of a new anti-cancer agent.

Berberine and RNAi-Targeting Telomerase Reverse Transcriptase (TERT) and/or Telomerase RNA Component (TERC) Caused Oxidation in Colorectal Cancer Cell Line, HCT 116: An Integrative Approach using Molecular and Metabolomic Studies.

Colorectal cancer (CRC) is the most common cancer in both men and women and is associated with increased telomerase levels and activity. The potential downstream effects of TERT and/or TERC downregulation by berberine (a telomerase inhibitor) or RNA interference (RNAi) on various target RNAs, proteins, relative telomerase activity (RTA), relative telomere length (RTL), hydrogen peroxide concentration [H2O2], percentage of cell cycle distribution, cell size and granularity as well as cellular metabolites were explored in HCT 116 cell line. Knockdown of TERT decreased TERC. The downregulation of TERT and/or TERC caused increment of [H2O2], G0/G1 phase arrest in addition to decreased S and G2/M phases, as well as diminished cell size. RTL was later reduced as a result of TERT, TERT and/or TERC downregulation which decreased RTA. It was discovered that xanthine oxidase (XO) was significantly and positively correlated at FDR-adjusted p value < 0.05 with RTA, TERT, TERT, TERC, and RTL. HCT 116 with decreased RTA was closely clustered in the Principal Component Analysis (PCA) indicating similarity of the metabolic profile. A total of 55 metabolites were putatively annotated in this study, potentially associated with RTA levels. The Debiased Sparse Partial Correlation (DSPC) Network revealed that RTA was directly correlated to TERT. There were 4 metabolic pathways significantly affected by low level of RTA which include (1) purine metabolism, (2) glycine, serine, and threonine metabolism, (3) glyoxylate and dicarboxylate metabolism, and (4) aminoacyl-tRNA biosynthesis. The Gene-Metabolite Interaction Network implied that reduced RTA level was related to the mechanism of oxidative stress. This study reveals the linkages between RTA to various selected RNAs, proteins, metabolites, oxidative stress mechanism and subsequently phenotypic changes in HCT 116 which is valuable to understand the intricate biological interactions and mechanism of telomerase in CRC.

Kadmiyum toksisitesi oluşturulan ratlarda arı sütünün farklı dozlarının oksidatif stres ve telomeraz enzimi üzerine etkileri

Purpose: Cadmium (Cd) is a toxic metal that seriously threatens human health due to environmental pollution, is widely used in industry and agriculture, and causes oxidative stress and tissue damage. This study aims to examine the effect of royal jelly (RJ) on oxidative status and telomerase enzyme activity in tissue damage induced by Cd.&#x0D; Materials and Methods: The experimental design was made with 6 rats in each group. A total of 6 groups were created: control group, Cd group, 250 mg/kg RJ group, Cd + 250 mg/kg RJ group, 400 mg/kg RJ group, Cd + 400 mg/kg RJ group. In the study, total oxidant status and total antioxidant status in blood serum were investigated by colorimetric method, and telomerase enzyme activity in ovarian tissue was investigated by ELISA method.&#x0D; Results: Cd caused an increase in oxidative capacity (23.80 ± 2.4) and a significant decrease was determined after RJ applications compared to the control group. After RJ application, the best total antioxidant response was observed in the 250 mg/kg RJ and Cd + 250 mg/kg RJ groups. Cd significantly reduced telomerase enzyme activity (0.90 ± 0.13). RJ administered for treatment after Cd application increased telomerase levels up to the control level (1.40 ± 0.05). The best treatment response was observed in the Cd + 250 mg/kg RJ group (1.42 ± 0.05).&#x0D; Conclusion: Cd causes oxidative stress and that RJ may have curative effects by increasing the antioxidant capacity and telomerase enzyme activity RJ is a promising natural product and can contribute to recovery.

Telomere length, telomerase reverse transcriptase promoter mutations, and melanoma risk.

Telomere repeats at chromosomal ends, critical for genomic integrity, undergo age-dependent attrition and telomere length has been associated with different disorders including cancers. In this study, based on 1469 patients and 1158 healthy controls, we show a statistically significant (P = 6 × 10-10 ) association between increased telomere length and melanoma risk. Mendelian randomization, using 5 telomere length-associated polymorphisms, ruled out confounding factors or reverse causality and showed association between increased telomere length and melanoma risk with odds ratio of 2.66 (95% confidence interval: 2.07-3.25). Age-dependent telomere attrition was faster in melanoma cases than controls (P = .01). The carriers of a highly penetrant germline -57A>C TERT promoter mutation, in a previously reported melanoma family, had longer telomeres than the noncarriers. The mutation causes increased TERT and telomerase levels through creation of a binding motif for E-twenty six (ETS) transcription factors and the carriers develop melanoma with an early age of onset and rapid progression to metastasis. In analogy, we hypothesize that increased telomere length in melanoma patients reflects stochastic increased telomerase levels due to common genetic variation. Paradoxically, we observed shorter telomeres (P = 1 × 10-5 ) in primary tumors from unrelated melanoma patients with (121) than without (170) somatic TERT promoter mutations that similar to the germline mutation, also create binding motifs for ETS transcription factors. However, the age-dependent telomere attrition was faster in tumors with the TERT promoter mutations than in those without such mutations. Besides a robust association between increased telomere length and risk, our data show a perturbed telomere homeostasis in melanoma.

Abstract 3408: Telomere length and TERT promoter mutations in cutaneous melanoma

Abstract Telomeres at chromosomal ends are comprised of multiple short repeat sequences. In humans TTAGGG repeats account for telomere length ranging 10-15 kb. Telomere sequences are mainly double stranded that end in a single stranded G-rich tail of 150-200 nucleotides. Telomeres in somatic cells undergo gradual shortening due to inherent limitations of DNA replication and limited levels of specialized enzyme telomerase that adds the repeats at chromosomal ends to maintain homeostasis. We previously reported somatic mutations in the core promoter of the telomerase reverse transcriptase (TERT) gene that lead to increased transcription of catalytic subunit and tumor specific telomerase reactivation. Telomere length per se is associated with risk in different cancers. In this study, we measured leukocyte telomere length using real-time PCR in 1469 melanoma patients and compared with that in 1158 matched healthy controls. The melanoma patients had statistically significantly longer telomeres than matched controls (t-test; P 6X10^-10). Mendelian randomization, carried out using two polymorphisms represented by rs1317082 and rs7726159 that associated with telomere length in genome wide association studies, showed association between increased telomere length and melanoma risk with an odds ratio of 2.3 (95% confidence interval 1.8-2.8). When measured in blood tissues from a melanoma family with the germline -57A&amp;gt;C TERT promoter mutation, the carriers had longer telomeres (median 1.12) than the non-carriers (median 0.87). The TERT promoter mutations create binding motifs for E-twenty six (ETS) transcription factors and in stem cell the presence of the promoter mutations resulted in continued TERT expression and telomerase activity following differentiation into adult cells. Individuals with the germline TERT promoter mutations develop melanoma with an early age of onset and rapid progression to metastases; two individuals in the family who lived past median age of onset developed several other malignancies, besides melanoma. Thus, dysregulated telomerase leads to a severe phenotype. In an analogy, we hypothesize that association of longer rather than shorter telomeres with an increased risk of melanoma reflects stochastic increased telomerase levels due to common genetic variation. In contrast, the telomere length was shorter in tumors from unrelated melanoma patients with (121) than without (170) somatic TERT promoter mutations (P 1X10^-5), which reflects the selection of the mutations at telomere crisis. Thus, a dynamic but controlled system evolved around telomere homeostasis when dysregulated leads to an increased cancer risk and affects tumor progression. Citation Format: Sivaramakirishna Rachakonda, Barbara Heidenreich, Eduardo Nagore, Rajiv Kumar. Telomere length and TERT promoter mutations in cutaneous melanoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3408. doi:10.1158/1538-7445.AM2017-3408

Endogenous Telomerase Reverse Transcriptase N-Terminal Tagging Affects Human Telomerase Function at Telomeres In Vivo.

Telomerase action at telomeres is essential for the immortal phenotype of stem cells and the aberrant proliferative potential of cancer cells. Insufficient telomere maintenance can cause stem cell and tissue failure syndromes, while increased telomerase levels are associated with tumorigenesis. Both pathologies can arise from only small perturbation of telomerase function. To analyze telomerase at its low endogenous expression level, we genetically engineered human pluripotent stem cells (hPSCs) to express various N-terminal fusion proteins of the telomerase reverse transcriptase from its endogenous locus. Using this approach, we found that these modifications can perturb telomerase function in hPSCs and cancer cells, resulting in telomere length defects. Biochemical analysis suggests that this defect is multileveled, including changes in expression and activity. These findings highlight the unknown complexity of telomerase structural requirements for expression and function in vivo.

Functional Assessment of Pharmacological Telomerase Activators in Human T Cells

Telomeres are structures at the ends of chromosomes that shorten during cell division and eventually signal an irreversible state of growth arrest known as cellular senescence. To delay this cellular aging, human T cells, which are critical in the immune control over infections and cancer, activate the enzyme telomerase, which binds and extends the telomeres. Several different extracts from the Astragalus membranaceus root have been documented to activate telomerase activity in human T cells. The objective of this research was to compare two extracts from Astragalus membranaceus, TA-65 and HTA, for their effects on both telomerase and proliferative activity of human CD4 and CD8 T cells. Our results demonstrate that, TA-65 increased telomerase activity significantly (1.3 to 3.3-fold relative to controls) in T cell cultures from six donors tested, whereas HTA only increased telomerase levels in two out of six donors. We also demonstrate that TA-65 activates telomerase by a MAPK- specific pathway. Finally, we determine that during a three-day culture period, only the T cells treated with the TA-65 extract showed a statistically significant increase in proliferative activity. Our results underscore the importance of comparing multiple telomerase activators within the same experiment, and of including functional assays in addition to measuring telomerase activity.

Suppression of Proliferative Defects Associated with Processing-defective Lamin A Mutants by hTERT or Inactivation of p53

Hutchinson-Gilford progeria syndrome (HGPS) is a rare, debilitating disease with early mortality and rapid onset of aging-associated pathologies. It is linked to mutations in LMNA, which encodes A-type nuclear lamins. The most frequent HGPS-associated LMNA mutation results in a protein, termed progerin, with an internal 50 amino acid deletion and, unlike normal A-type lamins, stable farnesylation. The cellular consequences of progerin expression underlying the HGPS phenotype remain poorly understood. Here, we stably expressed lamin A mutants, including progerin, in otherwise identical primary human fibroblasts to compare the effects of different mutants on nuclear morphology and cell proliferation. We find that expression of progerin leads to inhibition of proliferation in a high percentage of cells and slightly premature senescence in the population. Expression of a stably farnesylated mutant of lamin A phenocopied the immediate proliferative defects but did not result in premature senescence. Either p53 inhibition or, more surprisingly, expression of the catalytic subunit of telomerase (hTERT) suppressed the early proliferative defects associated with progerin expression. These findings lead us to propose that progerin may interfere with telomere structure or metabolism in a manner suppressible by increased telomerase levels and possibly link mechanisms leading to progeroid phenotypes to those of cell immortalization.

Telomerase activity levels in the surgical margin and tumour distant tissue of the squamous cell carcinoma of the head-and-neck.

The survival of patients with a head‐and‐neck squamous cell carcinoma is determined by loco‐regional recurrence and second primary carcinomas. As a complement to histopathology, molecular changes of tumour marginal and tumour distant tissue may confirm curative surgical tumour extirpation. We tested telomerase activity with PCR‐ELISA kits. 20 tumour margin biopsies were chosen by the surgeon from 20 patients. In addition, 3 tissue samples were taken from each of 20 additional patients, one from the carcinoma centre, the tumour margin and one distant from the tumour. 50% of the carcinoma centres were telomerase‐positive. Thirteen of the 40 tumour margin samples showed increased telomerase levels, and in 3 of these residual carcinoma was histopathologically detected. Six of the 20 tumour distant tissues revealed increased telomerase levels. Telomerase positivity in carcinoma‐free tumour margins correlated with a good prognosis. Confirmation of the results in a larger patient group is needed.