• Proceedings of the KSAR Conference
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• 2002.06a
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• pp.39-39
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• 2002

The mechanisms underlying the visual assessment and resulting in optimum embryonic development following in vitro maturation, fertilization, and culture are unclear. It is known that in vitro produced embryos show more frequent occurrence of fragmentation, which result in poor developmental potential and decreased implantation rate. The objective of this study was to investigate the apoptotic rates in in vitro fertilization (IVF) and nuclear transferred (NT)bovine blastocyst. (omitted)

• Biomedical Science Letters
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• v.8 no.3
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• pp.161-165
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• 2002

Apoptosis can be difficult to detect in routine histological sections. Since extensive DNA fragmentation is an important characteristic of this process, visualization of DNA breaks could greatly facilitate the identification of apoptotic cells. Several techniques for the qualitative and quantitative detection of this process have been established; recently, an in situ nick end-labelling technique based on the detection of DNA fragmentation, which is a molecular characteristic of apoptotic cell death, was described. Applying this method to paraffin sections of rat tissues, sensitivity was observed to be inconsistently low with regard to the expected number of apoptotic cells. I describe a new modified method for formalin-fixed, paraffin-embedded tissue sections, pretense pretreatment to permeate the tissue sections that involves an TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling) is acknowledged as a method of choice in the rapid identification and quantification of the apoptotic cell fraction in paraffin tissue preparations. TUNEL was performed without apoptosis and with apopotosis samples to each of the three concentrations of proteinase K (10, 25, 40 mg/ml) pretreatments. In this study, I show that chemical pretreatments of the tissue sections in proteinase K (25 mg/ml for 15 min at room temperature) considerably enhances the sensitivity of this nick end labelling technique.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.11
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• pp.5455-5461
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• 2012

Luteolin is a plant flavonoid which exhibits anti-oxidative, anti-inflammatory and anti-tumor effects. However, the antiproliferative potential of luteolin is not fully understood. In this study, we investigated the effect of luteolin on cell cycling and apoptosis in human esophageal squamous carcinoma cell line Eca109 cells. MTT assays showed that luteolin had obvious cytotoxicity on Eca109 with an $IC_{50}$ of $70.7{\pm}1.72{\mu}M$ at 24h. Luteolin arrested cell cycle progression in the G0/G1 phase and prevented entry into S phase in a dose- and time-dependent manner. as assessed by FCM. Luteolin induced apoptosis of Eca109 cells was demonstrated by AO/EB staining assay and annexin V-FITC/PI staining. Moreover, luteolin downregulated the expression of cyclin D1, survivin and c-myc, and it also upregulated the expression of p53, in line with the fact that luteolin was able to inhibit Eca109 cell proliferation.

• Journal of Dental Rehabilitation and Applied Science
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• v.23 no.1
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• pp.95-104
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• 2007

I. Objective The primary requirement of an endodontic root canal sealer is the biologic compatibility, because they remain in close contact with living periapical tissues over a long period of time. The aim of this study was the evaluation of cytotoxicity and genotoxicity of resin-based root canal sealers, AH 26 and ADSEAL. II. Material & Methods In this study, human periodontal ligament cells, human oral cancer cells (KB) and mouse osteoblasts (MC-3T3-E1) were used. Specimens of AH26, ADSEAL were eluted with culture medium for 1, 3, 5 and 7 days. Cytotoxicity was evaluated by using tetrazolium bromide reduction assay (MTT assay) for mitochondrial enzyme activity and cell viability. Genotoxicity was evaluated by using alkaline single cell gel electrophoresis assay (Comet assay). Also cell apoptosis induced by AH 26 was detected by Hoechst33258 staining. III. Results AH 26 and ADSEAL exhibited cytotoxic effects in all investigated cell groups. Genotoxicity was also noted for both sealers in mouse osteoblasts (MC-3T3-E1). But, ADSEAL presented significantly low cytotoxicity and genotoxicity compared with AH 26. Cytotoxicity and genotoxicity induced by AH 26 resulted in apopotosis. IV. Conclusion Our results clearly indicate that the recently invented ADSEAL has better biocompatibility than another resin based root canal sealer, AH 26. However ideal root canal sealer should have not only biocompatibility but also satisfactory physico-chemical properties such as sealing ability and stability. Thus continuous studies and developments should follow.

• Korean Journal of Veterinary Research
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• v.40 no.2
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• pp.221-228
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• 2000

We performed this study to determine the effect of Bu-Zhong-Yi-Qi-Tang, as a prescription of traditional Oriental medicine, and its major ingredients on jejunal crypt survival, endogenous spleen colony formation, apopotosis in jejunal crypt cells, lethality and hematological change of mice irradiated with high and low dose of Y-radiation. Bu-Zhong-Yi-Qi-Tang administration before irradiation protected the jejunal crypts (p<0.0001), increased the formation of endogenous spleen colony (p<0.05) and reduced the frequency of radiation-induced apoptosis (p<0.05). The survival rate and mean survival time of the groups treated with Bu-Zhong-Yi-Qi-Tang within 30 days after the treatment were far better than the irradiation control group. In the experiment on the effect of ingredients of Bu-Zhong-Yi-Qi-Tang, the result indicated that the extract of Rensan (Panax ginseng), Danggui (Angelica sinensis), Shengma (Cimicifuga heracleifolia) and Chaihu (Bupleurum falcatnosa) might have a major radioprotective effect. Although the mechanisms of this inhibitory effect remain to be elucidated, these results indicated that BU-Zhong-Yi-Qi-Tang might be a useful radioprotector, especially since it is a relatively nontoxic natural product. Further studies are needed to characterize better the protective nature of Bu-Zhong-Yi-Qi-Tang extract and its ingredients.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.6
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• pp.2251-2256
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• 2015

To study effects of cellular FLICE (FADD-like IL-$1{\beta}$-converting enzyme)-inhibitory protein (c-FLIP) inhibition by RNA interference (RNAi) on sensitivity of U2OS cells to tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL)-induced apoptosis, plasmid pSUPER-c-FLIP-siRNA was constructed and then transfected into U2OS cells. A stable transfection cell clone U2OS/pSUPER-c-FLIP-siRNA was screened from the c-FLIP-siRNA transfected cells. RT-PCR and Western blotting were applied to measure the expression of c-FLIP at the levels of mRNA and protein. The results indicated that the expression of c-FLIP was significantly suppressed by the c-FLIP-siRNA in the cloned U2OS/pSUPER-c-FLIP-siRNA as compared with the control cells of U2OS/pSUPER. The cloned cell line of U2OS/pSUPER-c-FLIP-siRNA was further examined for TRAILinduced cell death and apoptosis in the presence of a pan-antagonist of inhibitor of apoptosis proteins (IAPs) AT406, with or without 4 hrs pretreatment with rocaglamide, an inhibitor of c-FLIP biosynthesis, for 24 hrs. Cell death effects and apoptosis were measured by the methods of MTT assay with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and flow cytometry, respectively. The results indicated that TRAIL-induced cell death in U2OS/pSUPER-c-FLIP-siRNA was increased compared with control cells U2OS/pSUPER in the presence or absence of AT406. Flow cytometry indicated that TRAIL-induced cell death effects proceeded through cell apoptosis pathway. However, in the presence of rocaglamide, cell death or apoptotic effects of TRAIL were similar and profound in both cell lines, suggesting that the mechanism of action for both c-FLIP-siRNA and rocaglamide was identical. We conclude that the inhibition of c-FLIP by either c-FLIP-siRNA or rocaglamide can enhance the sensitivity of U2OS to TRAIL-induced apopotosis, suggesting that inhibition of c-FLIP is a good target for anti-cancer therapy.

• Journal of Life Science
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• v.18 no.6
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• pp.804-813
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• 2008

Cordyceps militaris, the Chinese medicinal fungal genus Cordyceps, is reported to possess many pharmacological activities including immunological stimulating, anti-cancer, anti-virus and anti-infection activities. However, the molecular mechanisms of C. militaris on biochemical actions in cancer have not been clearly elucidated yet. In the present study, we investigated the anti-proliferative activity of the water extract of C. militaris (WECM) in human hepatocellular carcinoma HepG2 cells. It was found that WECM could inhibit the cell growth in a dose-dependent manner, which was associated with morphological changes and apoptotic cell death such as formation of apoptotic bodies and increased populations of apoptotic sub-G1 phase. Apoptotic cell death of HepG2 cells by WECM was connected with a up-regulation of pro-apoptotic Bax expression, tumor suppressor p53 and cyclin-dependent kinase inhibitor p21 (WAF1/CIP1). In addition, WECM treatment induced the proteolytic activation of caspase-3 and a concomitant degradation and/or inhibition of poly (ADP-ribose) polymerase (PARP), ${\beta}-catenin$ and phospholipase $(PLC)-{\gamma}1$ protein. Furthermore, caspase-3 inhibitor, z-DEVD-fmk, significantly inhibited WECM-induced apoptosis demonstrating the important role of caspase-3 in the observed cytotoxic effect. Taken together, these findings provide important new insights into the possible molecular mechanisms of the anti-cancer activity of C. militaris.

• Journal of Applied Biological Chemistry
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• v.63 no.2
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• pp.147-152
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• 2020

Farrerol is a flavanone isolated from the traditional Chinese herb 'Man-shan-hong' (Rhododendron dauricum L.). Farrerol has been reported to have various bioactivities including anti-oxidant, anti-inflammation, and anti-fungal. However, anti-cancer effect of farrerol has not yet been reported in MCF-7 breast cancer cells. In the present study, we investigated the anti-cancer effect of farrerol on MCF-7 cells. Farrerol decreased viability and induced apoptosis of MCF-7 cells in a dose dependent manner. Ferrerol exhibited a significant anti-proliferation effect with a half-maximal inhibitory concentration (IC₅₀) values of 145.04±1.4 μM in MTT assay, when MCF-7 cells were treated with ferrerol for 48 h. Also, ferrerol induced apoptotic bodies of MCF-7 cells as evaluated by TUNEL assay and Annexin V/PI staining using FACS. By mechanism of action, ferrerol regulated the activation of p38 mitogen-activated protein kinase and altered the expression level of BAX, Bcl-2, and Poly ADP Ribose Polymerase in MCF-7 cells. In summary, our finding demonstrated that ferrerol has anti-cancer effect through regulating the activation and expression of apoptosis-related proteins in MCF-7 cells.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.6
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• pp.714-720
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• 2008

Cordyceps militaris is a medicinal fungus which has been used for patient suffering from cancer in Oriental medicine. It was previously reported that C. militaris extracts are capable of inhibiting tumor growth and inducing apoptosis; however, the anti-poliferative effects of human cancer cells have been poorly understood. In this study, to elucidate the anti-cancer mechanisms of human cancer cells by treatment with aqueous extract of C. militaris (AECM), we investigated the anti-proliferative effects of AECM in human hepatocarcinoma Hep3B cells. AECM treatment inhibited the growth of Hep3B cells and induced the apoptotic cell death in a concentration-dependent manner such as formation of apoptotic bodies and increased populations of apoptotic-sub G1 phase. The induction of apoptosis by AECM was connected with a proteolytic activation of caspase-3 and caspase-8. and concomitant degradation of poly (ADP-ribose) polymerase (PARP) and ${\beta}$-catenin proteins. Furthermore, caspase-3 inhibitor, z-DEVD-fmk, significantly inhibited AECM-induced apoptosis demonstrating the important role of caspase-3 in the bserved cytotoxic effect. Taken together, these findings suggest that AECM-induced inhibition of human hepatocarcinoma cell proliferation is associated with the induction of apoptotic cell death via activation of caspase-3 and C. militaris may have therapeutic potential in human cancer.

• Journal of the Korean Society of Food Science and Nutrition
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• v.31 no.3
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• pp.521-526
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• 2002

A number of experimental and epidemiological studies have implicated that antiestrogenic effects of estrogen-like compounds in legumes and plant seeds are responsible for lowering breast cancer risk in human. However, few studies have been conducted to illustrate the possible chemopreventive effects of Korean traditional food materials. This study was performed to determine the cytotoxic and apoptotic effects of yellow soybeans, black soybeans and brown rice extracts on hormone-dependent and hormone-independent human breast cancer cells. Methanol-or acetone-soluble fractions of soybeans or brown rice were incubated with hormone-dependent cells (MCF-7) or hormone-independent cells (MDA-MB-231). Cell cytotoxicity was measured by MTT assay at 24, 48 and 72 hrs of incubation. Apoptotic effects of these extracts toward breast cancer cells were also determined at 48 hrs of incubation by measuring DNA fragmentation. Results indicated that the acetone-soluble fraction of brown rice exerted strongest cytotoxic effect on MCF-7 ceIls, although other fractions also reduced the number of viable MCF-7 cells after 48 hrs of incubation. Both acetone and methanol soluble fractions of all samples exerted a significant cytotoxicity towards MDA-MB-231 cells after 24 hrs of incubation, and acetone and methanol soluble fractions of brown rice were especially effective in these cells. At 48 hrs of incubation, methanol fractions of all three samples induced apopotosis of MDA-MB-231 cells. These results indicate methaol or acetone soluble fractions of yellow soybeans, black soybeans and brown rice induce cytotoxicity in both hormone-dependent and hormone-independent breast cancer cells. Therefore, possible mechanisms of cell cytotoxicity do not necessarily include antiestrogenic effects of soybean or brown rice extract. A possible anticarcinogenic effect of brown rice methanol-soluble fraction may mediated through their apoptotic effect. Further studies are requried to elucidate responsible compounds and mechanisms involved in observed anticarcinogenesis.