• Asian Pacific Journal of Cancer Prevention
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• 제14권10호
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• pp.5911-5913
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• 2013

Cyclamen coum is a traditional medicinal plant in the Turkey. Its anticancer properties and whether cyclamen extract induces any cytotoxicity in solid cancer cell lines have not been thoroughly investigated previously. Therefore we examined cytotoxic effects on cervical cells; HeLa and non small cell lung cancer cell, H1299, lines; Cyclamen extract induced cellular death of both HeLa and H1299 cells in a dose dependent manner. We also analyzed the capacity of cyclamen extract to induce apoptosis by the TUNEL method. Here, for the first time we report that the extract of Cyclamen coum, an endemic plant for Turkey, Bulgaria, Georgia and the Middle East can induce cytotoxicity via apoptosis in HeLa and H1299 cells. These results imply that cyclamen extract can be further analyzed to potentially find novel anticancer compounds.

• Clinical and Experimental Reproductive Medicine
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• 제45권3호
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• pp.101-109
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• 2018

In an age when a small quantity of sperm can lead to pregnancy through in vitro fertilization or intracytoplasmic sperm injection, selecting healthy sperm is important. Sperm DNA fragmentation (SDF) is known to be higher in infertile men. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) and the alkaline comet test are SDF tests that directly measure DNA damage and have shown closer correlations with assisted reproduction results than indirect tools such as the sperm chromatin structure assay or the sperm chromatic dispersion test. It is difficult; however, to endorse a single test as the best test overall; instead, it is best to select a testing method based on each patient's clinical condition and goals. In a couple struggling with infertility, if the male partner has a high level of SDF, he should aim to decrease SDF through lifestyle modifications, antioxidant treatment, and ensuring an appropriate duration of abstinence, and physicians need to treat the underlying diseases of such patients. If sperm DNA damage continues despite the patient's and physician's efforts, other methods, such as micromanipulation-based sperm selection or testicular sperm extraction, should be used to select healthy sperm with nuclear DNA integrity.

• 한국동물생명공학회지
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• 제35권2호
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• pp.207-213
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• 2020

Cryopreservation is used for blastocyst preservation of most mammalian embryos and is an important technique for breeding. We aimed to compare the efficiency of the cryopreservation method using the standard Cryotop device and the ReproCarrier device, a domestic product manufactured in Korea. The efficacy of the two devices was analyzed based on the survival rate, intracellular levels of reactive oxygen species (ROS), and apoptosis of the vitrified bovine blastocysts. The survival rates of the vitrified-warmed blastocysts were similar between the ReproCarrier group (58.4 ± 17.7%) and Cryotop group (59.9 ± 14.1%). Intracellular ROS levels and apoptotic index were determined by DCFDA staining and TUNEL assay. Changes in intracellular ROS levels, number of total nuclei, and cellular apoptosis of vitrified blastocysts after cryopreservation were not significantly different between the two groups. These results indicate that the ReproCarrier device method is as effective as the standard Cryotop method for vitrification of bovine blastocysts in vitro.

• 한국식품위생안전성학회지
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• 제25권3호
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• pp.269-277
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• 2010

본 연구에서는 azoxymethane (AOM)과 dextran sodium sulfate (DSS)로 유도된 대장 발암과정에 대한 셀레늄의 방어 효과를 조사하였다. 셀레늄 결핍(0.02 ppm Se), 정상(0.1 ppm Se), 과다(0.5 ppm Se)사료를 12주간 식이로 급여하여 혈액검사와 대장암 발생의 초기단계인 aberrant crypt foci (ACF)수를 측정했으며, 암 발생율을 조사하였다. ICP-AES를 사용하여 간의 셀레늄 농도를 측정하였으며, 또한 셀레늄포함 항산화효소인 glutathione peroxidase (GPx) 활성을 알아보았다. 또한 TUNEL assay와 PCNA, $\beta$-catenin에 대한 면역조직 염색을 수행하였다. ACF 수 및 종양 발생률에 있어서, 셀레늄과다사료를 급여한 군이 정상셀레늄사료를 급여한 군보다 낮았으며, 셀레늄결핍사료를 급여한 군은 오히려 ACF 수 및 종양 발생률이 높았다. GPx 활성은 셀레늄의 섭취가 과다한 군에서 높게 나타났으며, 이 때, TUNEL에서 apoptotic positive cell이 증가하는 것을 확인했다. 또한 셀레늄의 섭취가 과다한 군에서 PCNA와 $\beta$-catenin의 발현이 감소됨을 볼 수 있었다. 본 마우스 모델실험에서 셀레늄은 여러 기전에 의해 대장암 발생을 억제할 수 있을 것으로 사료된다.

• The Korean Journal of Physiology and Pharmacology
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• 제1권6호
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• pp.817-824
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• 1997

Extracellular ATP elicits various biological responses and plays a significant role in physiological regulation. Recently, ATP-induced growth inhibitions were reported in some tumor cell lines, but these effects and mechanisms are not well hewn. This study was conducted to investigate ATP-induced growth inhibition in mouse $leukemic(P388D_1)$ cells. ATP inhibited cell growth in a dose-dependent manner as analyzed by MTS assay$(IC_{50}: 33.1\;{\mu}M)$. Nucleotides other than ATP, such as ADP$(37.5;{\mu}M)$ and AMP$(33.2;{\mu}M)$ had the same effects as ATP but adenosine$(57.8;{\mu}M)$ showed less effect than ATP. ATP attenuated the cells in $G_0/G_l\;and\;G_2/M$ phases but increased those in S phase in flow cytometric analysis. Hypodiploid cells$(A_0)$, the presumptive findings of apoptosis, were found among the ATP-treated cells. ATP induced DNA fragmentation into $180{\mu}200\;bps $as measured by electrophoresis. some apoptotic cells were stained by TUNEL method. ATP increased the intracellular free $Ca^{++}$concentration$([Ca^{++}]_i)$ and the increment of $([Ca^{++}]_i)$ was caused by influx from the extracellular space. These results suggest that extracellular ATP induces growth inhibition through apoptosis.

• 대한한방내과학회지
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• 제27권1호
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• pp.72-83
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• 2006

Objective : This study was designed to identify lipid protection formation in stratum corneum and anti-inflammatory effects of Sopungdojeok-tang(SD) on atopic dermatitis(AD). Materials and Methods : In Vivo, SD extract was orally administered to BALB/c mice at $2.5m{\ell}/kg/day$ for 2 days after 5% sodium dodecyl sulfate evoked atopic dermatitis in abdominal skin. Morphological changes were observed by immunohistochemical stain using monoclonal antibodies(BrdU, ceramide, MIP-2, $NF-{\kappa}B$ p50, IL-4, and STAT6) and TUNEL method. In vitro, the alterations of IL-4 mRNA expression were detected by RT-PCT in SD extract treated EL4 cells after phorbol-12-myristate-13-acetate and 4-tert-Octylphenol induce Th2 skewed condition. Results : SD is used in Oriental Medicine for its potential curative for atopic dermatitis. In this study, we have investigated the anti-inflammatory and lipid lamella repair effects of SD were investigated. SD decreased the number of eosinophil in atopic dermatitis induced mice. In the histological properties, the hyperplasia, edema, infiltration of lymphocytes, damage of intercellular space of stratum corneum, BrdU positive reacted cells in stratum basal, and degranulated mast cells and capillaries in dermal papillae decreased in mice with SD. Treatment of SD also decreased MIP-2, STAT6 and IL-4 in dermal papillae. The IL-4 mRNA expression decreased in a dose-dependant manner in SD treated EL4 cells. In addition, decrease of $NF-{\kappa}B$ p50 and increase of apoptotic cells in dermis were observed in SD treated mice. These data suggest that SD may beneficial for atopic dermatitis. Conclusions : These data suggest that SD is beneficial in treatment of atopic dermatitis, and that SD provides lipid protection in stratum corneum and anti-inflammatory effects on atopic dermatitis.

• Asian-Australasian Journal of Animal Sciences
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• 제24권9호
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• pp.1204-1210
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• 2011

Objective of this study was to examine the effect of sodium nitroprusside (SNP), a nitric oxide (NO) donor on steroid synthesis, growth and apoptosis of buffalo granulosa cells (GCs) in vitro. Follicular fluid of antral follicles (3-5 mm diameter) was aspirated and GCs were cultured in 0 (control), $10^{-3}$, $10^{-5}$, $10^{-7}$, $10^{-9}\;M$ of SNP for 48 h. To evaluate whether this effect was reversible, GCs were cultured in presence of $10^{-5}\;M$ SNP+1.0 mM $N^{\omega}$-nitro-L-arginine methyl ester (L-NAME) a NO synthase (NOS) inhibitor or hemoglobin (Hb, $1.0{\mu}g$) as NO scavenger. Nitrate/nitrite concentration was evaluated by Griess method, progesterone and estradiol concentrations by RIA and apoptosis by TUNEL assay. SNP ($10^{-3}$, $10^{-5}$, $10^{-7}\;M$) significantly (p<0.05) inhibited estradiol and progesterone synthesis, growth, disorganized GCs aggregates and induced apoptosis in a dose dependent manner. However, $10^{-9}\;M$ SNP induced the progesterone synthesis and stimulated GCs to develop into a uniform monolayer. Combination of SNP $10^{-5}$ M+L-NAME strengthened the inhibitory effect while, SNP+Hb together reversed these inhibitory effects. In conclusion, SNP at greater concentrations ($10^{-3}$, $10^{-5}$ and $10^{-7}\;M$) has a cytotoxic effect and it may lead to cell death whereas, at a lower concentration ($10^{-9}\;M$) induced progesterone synthesis and growth of GCs. These findings have important implications that NOS derived NO are involved at physiological level during growth and development of buffalo GCs which regulates the steroidogenesis, growth and apoptosis.

• 대한한방내과학회지
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• 제35권1호
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• pp.79-91
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• 2014

Objectives : To investigate the anti-cancer effect of Palbohoichoon-tang (PBHCT) extracts. Methods : The cell viability was assessed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MMT) assay and cell morphological changes were microscopically analyzed after staining with $10{\mu}M$ 2-[4-amidinophenyl]-6-indolecarbamidine dihydrochloride (DAPI) and TUNEL. We also analyzed expression of Bcl2, $Bcl_{xL}$, Bax, procaspase-3, procaspase-9, and procyclic acidic repetitive protein (PARP) by western blot method. Results : Observations showed that PBHCT induced the apoptotic cell death proved by increased sub-G1 phase cell population, apoptotic body formation and chromatin condensation. Western blot analysis of total cell lysates revealed that the PBHCT induced cleavage of caspase-9, caspase-3 and poly (ADP-ribose) polymerase (PARP). In addition, PBHCT dose-dependently increased the activity of caspase-9, caspase-3 and PARP-1. Furthermore, PBHCT reduced anti-apoptotic Bcl2, $Bcl_{xL}$ expression which contributed to the loss of mitochondrial membrane potential and the activations of caspase-9 and caspase-3. Conclusions : These findings suggest that PBHCT exerts anti-cancer effects on human neuroblastoma SH-SY5Y cells by inducing apoptotic death via down-regulation of anti-apoptotic proteins such as Bcl2 and $Bcl_{xL}$, up-regulation of pro-apoptotic proteins such as Bax, and activation of caspase cascades and PARP-1.

• Asian-Australasian Journal of Animal Sciences
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• 제18권9호
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• pp.1242-1248
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• 2005

This study was designed to investigate the in vitro developmental ability and apoptosis of bovine embryos nucleartransferred (NT) with frozen-thawed or cooled donor cells. Cultured adult bovine ear cells were used as donor cells after sub-culturing to confluence (CC), cooling to 4$^{\circ}C$ for 48 h, or freezing-thawing (FT). Apoptotic cells in blastocysts were evaluated for apoptosis by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) method. Fusion, cleavage and blastocyst rates were 69.0 (167/242), 68.8 (115/167), and 29.9 (50/167) with CC cells, 70.4 (88/125), 69.3 (61/88), and 29.6 (26/88) with cooled cells and 66.1 (117/177), 70.1 (82/117), and 13.7 (16/117) with FT cells, respectively. Blastocyst rates of NT embryos derived from FT cells were significantly lower than those from CC or cooled cells (p<0.05). In addition, NT blastocysts produced by using FT cells showed significantly higher apoptosis rates (6.4${\pm}$4.0%) than those produced by CC (2.8${\pm}$1.7%) or cooled (2.3${\pm}$1.3%) cells. However, cooling of donor cells had no significant adverse effect on blastocyst rate as well as apoptosis rate. Therefore, our results suggest that cooled cells may be used as an alternative to freshly cultured confluent culture cells, as donor cells, for the production of Somatic nuclear cloned cattle.

• Asian Pacific Journal of Cancer Prevention
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• 제15권12호
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• pp.4919-4923
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• 2014

Background: To investigate the effect of celecoxib on telomerase activity and apoptosis in a human laryngeal squamous carcinoma cell line (Hep-2 cells). Materials and Methods: The growth inhibition rate of Hep-2 cells in vitro was measured by MTT assay, and apoptosis by TUNEL assay and flow cytometry (FCM). The TRAP-ELISA method was used to determine telomerase activity in Hep-2 cells. The mRNA expression of human telomerase RNA component(hTR), human telomerase reverse transcriptase (hTERT) and human telomerase-associated protein(hTEP1) was determined by RT-PCR assay. Expression of Bax and Bcl-2 proteins was assessed by Western blotting. Results: Celecoxib can inhibit proliferation and induce apoptosis in a dose- and time-dependent manner, repress telomerase activity, decrease hTERT mRNA and Bcl-2 protein expression and increase Bax protein expression, PGE2 had no effect on telomerase. Conclusions: Celecoxib had the antiproliferative and pro-apoptotic effect in Hep-2 cells. Apoptosis was accompanied by a decrease in telomerase activity which was directly correlated with hTERT mRNA and up-regulation of Bax/Bcl-2. Bcl-2 may thus play an important role in telomerase activity as well as apoptosis.