• Nutrition Research and Practice
• /
• v.13 no.1
• /
• pp.58-63
• /
• 2019

BACKGROUND/OBJECTIVES: Telomeres are located at the chromosomal ends and progressively shortened during each cell cycle. Telomerase, which is regulated by hTERT and c-MYC, maintains telomeric DNA sequences. Especially, telomerase is active in cancer and stem cells to maintain telomere length for replicative immortality. Recently we reported that walnut phenolic extract (WPE) can reduce cell viability in a colon cancer stem cell (CSC) model. We, therefore, investigated the effect of WPE on telomere maintenance in the same model. MATERIALS AND METHODS: $CD133^+CD44^+$ cells from HCT116, a human colon cancer cell line, were sorted by Fluorescence-activated cell sorting (FACS) and treated with WPE at the concentrations of 0, 10, 20, and $40{\mu}g/mL$ for 6 days. Telomere lengths were assessed by quantitative real-time PCR (qRT-PCR) using telomere specific primers and DNA extracted from the cells, which was further adjusted with single-copy gene and reference DNA ($ddC_t$). Telomerase activity was also measured by qRT-PCR after incubating the PCR mixture with cell protein extracts, which was adjusted with reference DNA ($dC_t$). Transcriptions of hTERT and c-MYC were determined using conventional RT-PCR. RESULTS: Telomere length of WPE-treated cells was significantly decreased in a dose-dependent manner ($5.16{\pm}0.13$ at $0{\mu}g/mL$, $4.79{\pm}0.12$ at $10{\mu}g/mL$, $3.24{\pm}0.08$ at $20{\mu}g/mL$ and $3.99{\pm}0.09$ at $40{\mu}g/mL$; P = 0.0276). Telomerase activities concurrently decreased with telomere length ($1.47{\pm}0.04$, $1.09{\pm}0.01$, $0.76{\pm}0.08$, and $0.88{\pm}0.06$; P = 0.0067). There was a positive correlation between telomere length and telomerase activity (r = 0.9090; P < 0.0001). Transcriptions of both hTERT and c-MYC were also significantly decreased in the same manner. CONCLUSION: In the present cell culture model, WPE reduced telomere maintenance, which may provide a mechanistic link to the effect of walnuts on the viability of colon CSCs.

• Nutrition Research and Practice
• /
• v.11 no.4
• /
• pp.334-339
• /
• 2017

BACKGROUND/OBJECTIVES: Telomere length is a useful biomarker for determining general aging status. Some studies have reported an association between alcohol consumption and telomere length in a general population; however, it is unclear whether the alcohol flush reaction, which is an alcohol-related trait predominantly due to acetaldehyde dehydrogenase deficiency, is associated with telomere length. This cross-sectional study aimed to evaluate the associations of alcohol consumption and alcohol flush reaction with leukocyte telomere length (LTL). SUBJECTS/METHODS: The study included 1,803 Korean adults. Participants provided blood specimens for LTL measurement assay and reported their alcohol drinking status and the presence of an alcohol flush reaction via a questionnaire-based interview. Relative LTL was determined by using a quantitative polymerase chain reaction. Statistical analysis used multiple linear regression models stratified by sex and age groups, and potential confounding factors were considered. RESULTS: Age-specific analyses showed that heavy alcohol consumption (> 30 g/day) was strongly associated with a reduced LTL in participants aged ${\geq}65years$ (P < 0.001) but not in younger participants. Similarly, the alcohol flush reaction was associated with a reduced LTL only in older participants who consumed > 15 g/day of alcohol (P < 0.01). No significant alcohol consumption or alcohol flush reaction associations with LTL were observed in the sex-specific analyses. CONCLUSIONS: The results suggest that older alcohol drinkers, particularly those with the alcohol flush reaction, may have an accelerated aging process.

• Journal of Animal Science and Technology
• /
• v.53 no.3
• /
• pp.195-202
• /
• 2011

Maintenance of adequate telomere length in developing cells is the most important concern to preserve the integrity of the genome. The length of telomere is strictly regulated by numerous telomere-binding proteins and/or interacting factors. Even though the expression of telomerase in the male reproductive tract has been characterized, developmental expressional profiling of telomerase and other telomere-associated proteins has not been determined in detail. The present study was attempted to examine expression patterns of catalytic subunit (Tert) and RNA component (Terc) of telomerase and two telomerase associated factors, telomerase associated protein 1 (Tep1) and TERF1 (TRF1) interacting nuclear factor 2 (Tinf2) in the testis and seminal vesicle of male rat during postnatal development. The real-time PCR analysis was utilized to quantify mRNA expression of molecules. The abundance of Tep1 mRNA in the testis and seminal vesicle was the highest at 5 months of age. Expressional fluctuation of Tinf2 during postnatal development was found in the testis, while expression of Tinf2 in the seminal vesicle was gradually increased until 5 months of age and then significantly decreased later. mRNA level of Tert gene in the testis was significantly increased at the adult and the elder, while the highest expression of Tert gene in the seminal vesicle was found at 5 months of age. Expression of Terc transcript in the testis and seminal vesicle was the highest at 5 months of age, followed by significant reduction at 1 and 2 years of ages. Such differential gene expression of telomere-associated factors and telomerase components in different male reproductive tissues during postnatal development indicates that maintenance of telomere length would be regulated in tissue- and/or age-specific manners.

• Toxicological Research
• /
• v.19 no.3
• /
• pp.161-172
• /
• 2003

Explanted mammalian cells perform a limited number of cell division in vitro and than are arrested in a state known as replicative senescence. Such cells are irreversibly blocked, mostly in the G1 phase of cell cycle, and are no longer sensitive to growth factor stimulation. Thus replicative senescence is defined as a permanent and irreversible loss of replicative potential of cells. For this characteristic, replicative senescence seems to evolve to protect mammalian organism from cancer. However, senescence also contributes to aging. It seems to decrease with age of the cell donor and, as a form of cell senescence, is thought to underlie the aging process. Extensive evidence supports the idea that progressive telomere loss contributes to the phenomenon of cell senescence. Telomeres are repetitive structures of the sequence (TTAGGG)n at the ends of linear chromosomes. It has been shown that the average length of telomere repeats in human somatic cells decreases by 30∼200 bp with each cell division. It is generally believed that when telomeres reach a critical length, a signal is activated to initiate the senescent program. This has given rise to the hypothesis that telomeres act as mitotic clocks to regulate lifespan. One proposes that cumulative oxidative stress, mainly reactive oxygen species generated from mitochondria, may mainly cause telomere shortening, accelerating aging. Here, the biological importance and mechanism of replicative senescence were briefly reviewed. Also it was summarized that how oxidative stress affects replicative senescence and telomere shortening.

• Journal of Breast Cancer
• /
• v.21 no.4
• /
• pp.391-398
• /
• 2018

Purpose: The aim of this study was to investigate the association of telomere length with breast cancer risk. We simultaneously explored the association between telomerase reverse transcriptase gene polymorphisms and telomere length. Methods: We used real-time quantitative polymerase chain reaction to measure relative telomere length (RTL) in genomic DNA extracted from peripheral blood from 183 breast cancer cases and 191 healthy controls. Genotyping was performed using the Sequenom MassARRAY platform. Results: Our results show that breast cancer patients had significantly shorter RTLs than control subjects (p<0.05). When the RTLs were categorized into tertiles, we found that the lowest RTL was significantly associated with increased breast cancer risk compared with the highest RTL (odds ratio [OR], 2.33; 95% confidence interval [CI], 1.40-3.90; p=0.001). Subgroup analyses indicated that risk of breast cancer was also significantly increased in the lowest RTL compared with the highest RTL in age >40 years (OR, 2.41; 95% CI, 1.31-4.43;p=0.005), body mass index ${\leq}24kg/m^2$ (OR, 2.81; 95% CI, 1.55-5.10; p=0.001), and postmenopausal women (OR, 3.94; 95% CI, 1.63-9.51; p=0.002), respectively. In addition, individuals with the AA genotype of rs2853677 have longer telomeres than those of breast cancer patients with the AG genotype (p=0.011). Conclusion: Our results suggest that shorter RTL was associated with an increased risk of breast cancer. An association was found between the AA genotype of rs2853677 and longer RTLs in the case group. Functional studies are warranted to validate this association and further investigate our findings.

• IMMUNE NETWORK
• /
• v.2 no.4
• /
• pp.183-188
• /
• 2002

Gradual attrition of telomere to a critical short length elicits successive cellular response of cellular senescence and crisis. Cancer cells evade this process by maintaining functional telomeres via one of two known mechanisms of telomere maintenance. The first and most frequent mechanism involves reactivation of enzyme activity of telomerase, a ribonucleoprotein complex mainly via transcriptional up-regulation of TERT, a catalytic subunit of telomerase complex. The second mechanism utilizes telomerase-independent way termed ALT (for Alternative Lengthening of Telomere), which possibly involves recombination pathways. Thus master key for cellular immortalization is supposed to possess adequate telomere reserves. Indeed, telomerase can alone induce the immortalization under culture on feeder cell layers without generally known inactivation mechanism of tumor suppressor genes. Including this phenomena, this review will focus on telomerase and telomere-associated proteins, thereby implication of these proteins for cellular immortalization processes.

• Asian Pacific Journal of Cancer Prevention
• /
• v.14 no.2
• /
• pp.795-799
• /
• 2013

Ku70/80 heterodimer is a central element in the nonhomologous end joining (NHEJ) DNA repair pathway, Ku80 playing a key role in regulating the multiple functions of Ku proteins. It has been found that the Ku80 protein located at telomeres is a major contributor to radiosensitivity in some telomerase positive human cancer cells. However, in ALT human osteosarcoma cells, the precise function in radiosensitivity and telomere maintenance is still unknown. The aim of this study was to investigate the effects of Ku80 depletion in the U2OS ALT cell line cell line. Suppression of Ku80 expression was performed using a vector-based shRNA and stable Ku80 knockdown in cells was verified by Western blotting. U2OS cells treated with shRNA-Ku80 showed lower radiobiological parameters (D0, Dq and SF2) in clonogenic assays. Furthermore, shRNA-Ku80 vector transfected cells displayed shortening of the telomere length and showed less expression of TRF2 protein. These results demonstrated that down-regulation of Ku80 can sensitize ALT cells U2OS to radiation, and this radiosensitization is related to telomere length shortening.

• Asian Pacific Journal of Cancer Prevention
• /
• v.16 no.7
• /
• pp.2975-2979
• /
• 2015

Background: Telomere length is closely associated with cellular radiosensitivity and WRAP53 is required for telomere addition by telomerase. In this research we assessed radiosensitivity of laryngeal squamous cell carcinoma Hep-2 cell lines after WRAP53 inhibition, and analyzed the molecular mechanisms. Materials and Methods: phWRAP53-siRNA and pNeg-siRNA were constructed and transfected into Hep-2 cells with lipofectamine. Expression of WRAP53 was analyzed by RT-PCR and Western-blottin, radiosensitivity of Hep-2 cells was assessed colony formation assay, and the relative length of telomeres was measured by QPCR. Results: The data revealed that the plasmid of phWRAP53-siRNA was constructed successfully, and the mRNA and protein levels of WRAP53 were both obviously reduced in the Hep-2 cell line transfected with phWRAP53-siRNA. After Hep-2 cells were irradiated with X-rays, the $D_0$ and $SF_2$ were 2.481 and 0.472, respectively, in the phWRAP53-siRNA group, much lower than in the control group ($D_0$ and $SF_2$ of 3.213 and 0.592) (P<0.01). The relative telomere length in the phWRAP53-siRNA group was $0.185{\pm}0.01$, much lower than in the untreated group ($0.523{\pm}0.06$) and the control group ($0.435{\pm}0.01$). Conclusions: Decreasing the expression of WRAP53 using RNA interference technique can enhance the radiosensitivity of Hep-2 cell lines by influencing the telomere length. WRAP53 is expected to be a new target to regulate the radiosensitization of tumor cells.

• Proceedings of the SCSK Conference
• /
• 2003.09a
• /
• pp.73-84
• /
• 2003

Skin aging process on pre-menopausal women is a problem that needs to be prevented as early as possible. The decrease of oestrogen level which is one of the intrinsic factors of the skin aging process will affect the skin biological process, due to oestrogen receptors on the skin. A number of researches conducted on pre-menopausal women with the allocation of oestrogen hormone resulted in delaying the skin aging process. The administration of soy isoflavon, a phytoestrogen found in daily food, on pre-menopausal women is hoped to be able to prevent skin aging process, even clinically or molecular biologically. This research aims to explain the benefit of administering of soy isoflavon on skin aging process. The design of the research is randomised controlled trial (RCT). As many as 60 pre-menopausal women were collected with simple random sampling method. Soy isoflavon is an independent variable, while skin aging process is a dependent variable assessed from the hydration, sebum level, average roughness, depth of wrinkles, skin clarity, length of the telomere. Analysis was conducted using t and MANDVA tests and.the result showed a significance (F = 10,439; p = 0,001) over the allocation of soy isoflavon to the whole variable dependent, including the telomere length and the skin hydration, meant that allocation of soy isoflavon could delay skin aging process.

• Asian-Australasian Journal of Animal Sciences
• /
• v.23 no.4
• /
• pp.437-443
• /
• 2010

To be economically profitable, the poultry industry demands an increase in stocking density, which could adversely affect chicken welfare. The current study was performed to investigate the effect of stocking density on stress-related, heat shock protein genes (HSP70 and HSP90), 3-hydroxyl-3-methyl-glutaryl coenzyme A reductase (HMGCR) gene and telomere length in broiler chickens. Seven-day-old broiler chickens were housed at High (0.0578 $m^2$/bird), Standard (0.077 $m^2$/bird) and Low (0.116 $m^2$/bird) stocking densities with 8 replicates each until 35 d of age. The growth performance, such as body weight gain and average daily feed intake, was found to be significantly (p<0.05) higher in the Low density group, but these parameters did not show any difference between the High and Standard groups. Other growth performance, such as feed conversion ratio and final feed intake, showed no difference among the treated groups. The expression levels of HSP70 and HMGCR were found to be elevated with the increase of stocking density. The expression level of these genes was significantly (p<0.05) higher in the High density stocked group compared with the other groups, whereas the expression levels were not significantly different between the Low and Standard groups. The expression levels of HSP90 did not show any significant changes among the treated groups. The telomeric length of the birds housed in High density was reduced significantly (p<0.05) when compared to that of the birds in Low density. These results clearly indicate that birds stocked at high density show physiological adaptive changes indicative of stress at gene transcriptional and telomere levels.