• Journal of Life Science
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• v.18 no.1
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• pp.120-128
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• 2008

The death ligand tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)/ Apo1L is a cytokine that activates apoptosis through cell surface death receptors. TRAIL has sparked growing interest in oncology due to its reported ability to selectively trigger cancer cell death. Euphorbiae humifusae Wind has been used in traditional Oriental medicine as a folk remedy used for the treatment of cancer. However, the mechanism responsible for the anticancer effects of E. humifusae not clearly understood. Here, we show that treatment with subtoxic doses of water extract of E. humifusae (WEEH) in combination with TRAIL induces apoptosis in TRAIL-resistant human gastric carcinoma AGS cells. Combined treatment with WEEH and TRAIL induced chromatin condensation and sub-G1 phase DNA content. These indicators of apoptosis were correlated with the induction of caspase activity that resulted in the cleavage of poly (ADP-ribose) polymerase. Combined treatment also triggered the loss of mitochondrial membrane potential. Furthermore, co-treatment with WEEH and TRAIL down-regulated the protein levels of the anti-apoptotic proteins such as Bcl-2, Bcl-xL, XIAP and cIAP-1. Although more study will be needed to examine the detailed mechanisms, this combined treatment may offer an attractive strategy for safely treating gastric adenocarcinomas and the results provide important new insights into the possible molecular mechanisms of the anticancer activity of E. humifusae.

• Journal of Life Science
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• v.23 no.3
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• pp.389-398
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• 2013

Platycodin D is a major constituent of triterpene saponins, which is found in the root of Platycodon grandiflorum, Platycodi Radix, which is widely used in traditional Oriental medicine for the treatment of many chronic inflammatory diseases. Several pharmacological effects of this compound have been reported recently, such as anti-inflammation, immunogenicity, anti-adipogenesis, lowered cholesterol, and anti-cancer activity. However, the mechanism by which this action occurs is poorly understood. In this study, we found that platycodin D greatly increased the potential of the anti-proliferative effect in various cancer cell lines. Our data revealed that platycodin D treatment resulted in a time- and concentration-response growth inhibition of U937 cells by inducing apoptosis, as evidenced by the formation of apoptotic bodies, chromatin condensation, and the accumulation of cells in the sub-G1 phase. Apoptosis induction of U937 cells by platycodin D correlated with an increase in the Bax/Bcl-2 ratio and caused the down-regulation of IAP family members. In addition, platycodin D treatment resulted in proteolytic activation of caspase-3, the concomitant degradation of poly(ADP-ribose) polymerases, and the collapse of the mitochondria membrane potential (${\Delta}{\Psi}_m$). However, the cytotoxic effects induced by platycodin D treatment were significantly inhibited by z-DEVD-fmk, a caspase-3 inhibitor, which demonstrated the important role that caspase-3 played in the observed cytotoxic effect. These findings suggest that platycodin D may be a potential chemotherapeutic agent for use in the control of human leukemia U937 cells. These findings also provided important new insights into possible molecular mechanisms of the anti-cancer activity of platycodin D.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.7
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• pp.975-982
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• 2015

Piceatannol (trans-3,4,3',5'-trihydroxystilbene), a natural stilbene, is an analogue of resveratrol. In the present study, possible mechanisms by which piceatannol exerts its pro-apoptotic action in cultured human oral cancer YD-15 cells were investigated. To investigate whether or not piceatannol has effects on cancer cell viability, human oral YD-15 cells were treated with piceatannol (0, 50, and $100{\mu}M$). Piceatannol treatment ($100{\mu}M$) showed the strongest inhibition of cell proliferation and reduced cell viability in a dose-dependent manner. Chromatin condensation detected by DAPI staining significantly increased in a concentration-dependent manner, indicating apoptosis. Piceatannol treatment activated initiator Bax (pro-apoptotic) and cPARP in a concentration-dependent manner. Further, piceatannol induced down-regulation of Bcl-2 (anti-apoptotic). We also evaluated the activity of piceatannol against oral cavity cancer tumors in mice. Piceatannol-treated nude mice bearing YD-15 xenograft tumors exhibited significantly reduced tumor volume and weight due to the potent effect of piceatannol on tumor cell apoptosis, as determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay. Immunohistochemistry staining showed elevated expression of cleaved-caspase-3 as well as reduced expression of Ki-67 in the piceatannol-treated group. Therefore, piceatannol can be developed as a cancer preventive medicine due to its growth inhibitory effects and induction of apoptosis in human oral cancer cells.

• KSBB Journal
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• v.24 no.4
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• pp.403-409
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• 2009

H2AX, a crucial component of chromatin, is implicated in DNA repair, cell cycle check point and tumor suppression. The aim of this study was to identify direct binding partners of H2AX to regulate cellular responses to above mechanisms. Literature reviews and bioinformatical tools were attempted intensively to find binding partners of H2AX, which resulted in identifying two potential proteins, breast cancer-1 (BRCA1) and BRCA1-associated RING domain 1 (BARD1). Although it has been reported in vivo that BRCA1 co-localizes with H2AX at the site of DNA damage, their biochemical mechanism for H2AX were however only known that the complex monoubiquitinates histone monomers, including unphosphorylated H2AX in vitro. Therefore, it is important to know whether the complex directly interacts with H2AX, and also which regions of these are specifically mediated for the interaction. Using in vitro GST pull-down assay, we present here that BRCA1 and BARD1 directly bind to H2AX. Moreover, through combinational approaches of domain analysis, fragment clonings and in vitro binding assay, we revealed molecular details of the BRCA1-H2AX and BARD1-H2AX complex. These data provide the potential evidence that each of the BRCA1 nuclear localization signal (NLS) and BARD1 BRCA1 C-terminal (BRCT) repeat domain is the novel mediator of H2AX recognition.

• Journal of Embryo Transfer
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• v.18 no.1
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• pp.69-73
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• 2003

This study was undertaken to investigate the effect of three different porcine IVM media, TCM-199, mSOF and NCSU-23 on early development of porcine spontaneous parthenogenotes. Spontaneous parthenogenotes were induced by a prolonged culture of porcine oocytes in each medium. In Experiment 1, oocytes derived from gilts were matured in three IVM media and maturation of oocytes was evaluated by the status of chromatin configuration. Oocytes matured in mSOF, NCSU-23 and TCM-199 showed no significant difference (P>0.05) in maturation. Maturation rates at 48h after IVM were 83.1$\pm$2% (mSOF), 78.0$\pm$3% (NCSU-23) and 83.5$\pm$2% (mSOF), 78.0$\pm$3% (NCSU-23) and 83.5$\pm$2% (TCM-199). In Experiment 2, pronucleus formation and development to 6~8 cell stage of pig oocytes activated spontaneously. Pronucleus formation, cleavage rate and development to 6~8 cell embryos of porcine spontaneous parthenogenotes were assessed at 55~58 h, 96 h and 168h after IVM, respectively. Pronucleus formation (5.4$\pm$2% and 3.7 $\pm$ 1% vs 1.7 $\pm$ 3%) and development to the 6$\pm$8 cell (3.2$\pm$3% and 4.0$\pm$1% vs 1.4$\pm$3%) was significantly (P<0.05) higher in mSOF or NCSU-23 than TCM-199. In conclusion, the present study showed that oocytes matured in mSOF and NCSU-23 were spontaneously activated with higher frequency.

• Journal of Life Science
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• v.29 no.9
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• pp.1002-1009
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• 2019

Early life stress (ELS) increases the risk of depression. ELS may be involved in the susceptibility to subsequent stress exposure during adulthood. We investigated whether epigenetic mechanisms of p11 promoter affect the vulnerability to chronic unpredictable stress (CUS) induced by the maternal separation (MS). Mice pups were separated from their dams (3 hr/day from P1-P21). When the pups reached adulthood, we applied CUS (daily for 3 weeks). The levels of hippocampal p11 expression were analyzed by quantitative real-time PCR. The levels of acetylated and methylated histone H3 at p11 promoter were measured by chromatin immunoprecipitation. Depression-like behavior was measured by the forced swimming test (FST). The MS and CUS group exhibited significant decreases in p11 mRNA level and the MS plus CUS group had a greater reduction in this level than the CUS group. The MS plus CUS group also resulted in greater reduction in H3 acetylation than the CUS group. This reduction was associated with an upregulation of histone deacetylase 5. Additionally, the MS plus CUS group showed a greater decrease in H3K4met3 level and a greater increase in H3K27 met3 level than the CUS group. Consistent with the reduction of p11 expression, the MS plus CUS group displayed longer immobility times in the FST compared to the control group. Mice exposed to MS followed by CUS had much greater epigenetic alterations in the hippocampus compared to adult mice that only experienced CUS. ELS can exacerbate the effect of stress exposure during adulthood through histone modification of p11 gene.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.31 no.4
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• pp.599-607
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• 1998

To clarify annual reproductive cycle of Korean dark sleeper, Odontobutis platycephala (Iwata et Jeon), we examined the seasonal changes of gonadosomatic index (GSI), the proportional frequency of oocyte development stages in the ovary and the changes of sex steroid hormone levels in blood from December 1995 to November 1997. In July and August, GSI was 0.35 to 0.72 and most oocytes in the ovary were chromatin-nucleolus stage and perinucleolar stage (proportional frequency: $87\%\~96\%$). In September, GSI was 1.20 $\pm$ 0.12, some oocytes in the ovary were yolk vesifle stage (proportional frequency: $22.8\%$) and vitellogenic stage which appeared very rarely(proportional frequency: $2.2\%$). GSI increased gradually from October and reached 4.59± 0.61 to December. During this period, oocytes of vitellogenic stage increased slightly (proportional frequency in December: $22.1\%$). In January, GSI was 4.32 $\pm$ 0.72 but the proportional frequency of oocytes in vitellogenic stage increased (proportional frequency: $51.2\%$). from February, GSI was increased sharply and reached to 10.51 $\pm$ 1.04 in March, the highest value throughout the year and the proportional frequency of oocytes in vitellogenic stage also reached the highest levels (proportional frequency: $60\%$). From April, GSI was gradually decreased and fell down to 1.11 $\pm$ 0.35 in June. During this period, the proportional frequency of mature oocytes was the highest in April (proportional frequency of mature oocyte stage: $40\%$ in April, $12\%$ May, $5\%$ June) throughout the year, and atretic ovarian follicles were appeared. The blood level of estradiol-17$\beta$ ($E_2$), which stimulates the hepatic synthesis and secretion of vitellogenin, was $0.84{\pm}0.20\;ng/m{\ell}$ in August, and thereafter was not changed until December. from January, it increased sharply and reached the highest level of $ 2.85{\pm}0.35\;ng/m{\ell}$ in March throughout the year, but fell to $0.14{\pm}0.02\;ng/m{\ell}$ in July(P<0.05), 17$\alpha$-hydroxprogesterone(17$\alpha$-OHP) was the peak $13.37{\pm}0.52ng/m{\ell}$ in March, but no significant changes in other period(below $3ng/m{\ell}$, P<0.05). 17$\alpha$, 20$\beta$-dihydroxy-4-pregnen-3-one(17$\alpha$, 20$\beta$-P), which was known as the final maturation inducing hormone in teleost, was $0.74{\pm}0.09ng/m{\ell}$ in April and $0.54{\pm}0.07ng/m{\ell}$ in May, but no significant changes in other period (below $0.26\;ng/m{\ell}$, p<0.05). Taken together these results, the annual reproductive cycle of O. platycephala divided into 4 periods as follows: 1) ripe and spawning period from April to June, main spawning period was from April to May, 2) Resting period from July to August, 3) Growing period from September to December, 4) Maturing period from January to March. Moreover, It was showed that the changes of sex steroid hormone in blood played a important roles in the annual reproductive cycle of O. platycephala.