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Extract of Saccharina japonica Induces Apoptosis companied by Cell Cycle Arrest and Endoplasmic Reticulum Stress in SK-Hep1 Human Hepatocellular Carcinoma Cells

  • Jung, Hyun Il;Jo, Mi Jeong;Kim, Hyung-Rak;Choi, Yung Hyun;Kim, Gun-Do
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.7
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    • pp.2993-2999
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    • 2014
  • Saccharina japonica is a family member of Phaeophyceae (brown macro-alga) and extensively cultivated in China, Japan and Korea. Here, the potential anti-cancer effect of n-hexane fraction of S. japonica was evaluated in SK-Hep1 human hepatocellular carcinoma cells. The N-hexane fraction reduced cell viability and increased the numbers of apoptotic cells in a both dose- and time-dependent manner. Apoptosis was activated by both caspase-dependent and independent pathways. The caspase-dependent cell death pathway is mediated by cell surface death receptors and activated caspase-8 amplified the apoptotic signal either through direct activation of downstream caspase-3 or pro-apoptotic proteins (Bad, Bax and Bak) subsequently leading to the release of cytochrome c. On the other hand, caspase-independent apoptosis appeared mediated by disruption of mitochondrial membrane potential and translocation of AIF to the nucleus where they induced chromatin condensation and/or large-scale DNA fragmentation. In addition, the n-hexane fraction induced endoplasmic reticulum (ER)-stress and cell cycle arrest. The results suggested that potential anti-cancer effects of n-hexane extract from S. japonica on SK-Hep1 cells.

The Cytotoxic and Anti-proliferative Effects of 3-Hydrogenkwadaphnin in K562 and Jurkat Cells Is Reduced by Guanosine

  • Moosavi, Mohammad Amin;Yazdanparast, Razieh;Sanati, Mohammad Hasan
    • BMB Reports
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    • v.38 no.4
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    • pp.391-398
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    • 2005
  • 3-hydrogenwadaphnin (3-HK) is a new daphnane-type diterpene ester isolated from Dendrostellera lessertii with strong anti-tumoral activity in animal models and in cultures. Here, prolonged effects of this new agent on proliferation and viability of several different cancerous cell lines were evaluated. Using [$^3H$]thymidine incorporation, it was found that the drug inhibited cell proliferation and induced G1/S cell cycle arrest in leukemic cells 24 h after a single dose treatment. The cell viability of Jurkat cells was also decreased by almost 10%, 31% and 40% after a single dose treatment (7.5 nM) at 24, 48 and 72 h, respectively. The drug-treated cells were stained with acridine orange/ethidium bromide to document the chromatin condensation and DNA fragmentation. These observations were further confirmed by detection of DNA laddering pattern in the agarose gel electrophoresis of the extracted DNA from the treated cells. Treatment of K562 cells with the drug at 7.5, 15 and 30 nM caused apoptosis in 25%, 45% and 65% of the cells, respectively. Exogenous addition of $25-50\;{\mu}M$ guanosine and/or deoxyguanosine to the cell culture of the drug-treated cells restored DNA synthesis, released cell arrest at G1/S checkpoint and decreased the apoptotic cell death caused by the drug. These observations were not made using adenosine. However, the drug effects on K562 cells were potentiated by hypoxanthine. Based on these observations, perturbation of GTP metabolism is considered as one of the main reasons for apoptotic cell death by 3-HK.

Cytotoxic and Apoptotic Activites of Echinomycin Derivative (Echinomycin-7) on P388 Murine Leukemia Cells

  • Jeon, Hyang;Kim, Sung-Su;Kim, Yoon-Suk;Park, Yil-Sung;Kim, Yong-Hae;Choi, Sun-Ju;Kim, Soo-Kie;Kim, Tae-Ue
    • BMB Reports
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    • v.31 no.6
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    • pp.560-564
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    • 1998
  • Echinomycin-7 is an echinomycin derivative, Smethylated sulfonium perchlorate of echinomycin. We studied the in vitro cytotoxicity and in vivo antitumor activity of echinomycin-7 against P388 leukemia cells and compared the results with echinomycin. With respect to the cytotoxic effects, echinomycin-7 had cell line-dependent $IC_{50}$ values while echinomycin had similar values to several tumor cell lines. Also, in vivo antitumor activities were observed in tumor-bearing mice treated with both agents, which showed that echinomycin-7 had a broad therapeutic dose range. We also observed the apoptosis on leukemia cells treated with echinomycin-7 which exihibited the ladder pattern of DNA on electrophoresis. In addition to apoptosis, echinomycin-7 arrested $G_1/S$ phases of the cell cycle at the same time. We then examined the signaling pathway of echinomycin-7-induced apoptosis and showed that ERK of the MAP kinase family was activated and translocated into the nucleus by echinomycin-7 stimulation. This study suggests that echinomycin-7 acts as an antitumor agent through in vitro cytotoxicity and has in vivo antitumor activity against leukemia cells, and that the echinomycin-7- induced apoptosis might involve signal transduction via MAP kinases.

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Ethanol-eluted Extract of Rhus verniciflua Stokes Inhibits Cell Growth and Induces Apoptosis in Human Lymphoma Cells

  • Lee, Jeong-Chae;Kim, Ju;Jang, Yong-Suk
    • BMB Reports
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    • v.36 no.4
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    • pp.337-343
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    • 2003
  • Rhus verniciflua Stokes (RVS) has been used as a traditional herbal medicine. Several earlier studies indicated that an ethanol extract of RVS has both anti-oxidant and anti-tumor properties, although the mechanism for the activity remains to be elucidated. In this report, we prepared a highly purified ethanol extract from RVS, named REEE-1 ($\underline{R}$hus $\underline{e}$thanol $\underline{e}$luted $\underline{e}$xtract-1), and investigated the mechanism involved in its growth-inhibitory effect on the human B and T lymphoma cell lines, BJAB and Jurkat, respectively. Results from tritium uptake proliferation assays showed that the proliferative capacities of both BJAB and Jurkat cells were strongly suppressed in the presence of REEE-1. This was further confirmed through trypan blue exclusion experiments that revealed a dose-dependent decrease in viable cell numbers after REEE-1 treatment. REEE-1-mediated suppression of cell growth was verified to be apoptotic, based on the increase in DNA fragmentation, low fluorescence intensity in nuclei after propidium iodide staining, and the appearance of DNA laddering. In particular, REEE-1 exerted its anti-oxidant activity through the inhibition of hydroxyl radical-mediated degradation by iron ion chelation rather than direct scavenging of hydroxyl radicals. Furthermore, REEE-1 was revealed to be a potential scavenger of superoxide anions. Collectively, our findings suggest that REEE-1 is a natural anti-oxidant that could be used as a cancer chemo-preventive and therapeutic agent.

cAMP antagonizes ERK-dependent antiapoptotic action of insulin

  • Cui, Zhi Gang;Hong, Na-Young;Guan, Jian;Kang, Hee-Kyoung;Lee, Dae-Ho;Lee, Young-Ki;Park, Deok-Bae
    • BMB Reports
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    • v.44 no.3
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    • pp.205-210
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    • 2011
  • Insulin has antiapoptotic activity in various cell types. However, the signaling pathways underlying the antiapoptotic activity of insulin is not yet known. This study was conducted to determine if cAMP affects the antiapoptotic activity of insulin and the activity of PI3K and ERK in CHO cells expressing human insulin receptors (CHO-IR). Insulin-stimulated ERK activity was completely suppressed by cAMP-elevating agents like as pertussis toxin (Ptx) and cholera toxin (Ctx) after 4 h treatment. Insulin-stimulated PKB/Akt activity was not affected at all. Ptx treatment together with insulin increased the number of apoptotic cells and the degree of DNA fragmentation. Ctx or 8-br-cAMP treatment also increased the number of apoptotic cells and stimulated the cleavage of caspase-3 and the hydrolysis of PARP. Taken together, cAMP antagonizes the antiapoptotic activity of insulin and the main target molecule of cAMP in this process is likely ERK, not PI3K-dependent PKB/Akt.

Quality Improvement of Frozen and Chilled Beef biceps femoris with the Application of Salt-bicarbonate Solution

  • Sultana, A.;Nakanishi, A.;Roy, B.C.;Mizunoya, W.;Tatsumi, R.;Ito, T.;Tabata, S.;Rashid, H.;Katayama, S.;Ikeuchi, Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.6
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    • pp.903-911
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    • 2008
  • The effects of salt and bicarbonate solution on overall meat quality in beef biceps femoris muscle were investigated with the application of chilling and freezing conditions. Muscles were injected to a target of 120% of original meat weight with a solution containing 1.2 M sodium chloride, 0.25 M sodium bicarbonate and 0.1% ascorbic acid (pH 7.2). Half of the meat samples, considered as chill treatment and chill control, were stored at $4^{\circ}C$ up to five days; while the other half, frozen treatment and frozen control, were kept in a freezer at $-20^{\circ}C$ for seven days. Compared with untreated control, treated meats had higher water holding capacity (p<0.05), lower drip loss (p<0.05) and lower shear force (p<0.07) with higher overall acceptability (p<0.05) in sensory evaluation. Morphological observations demonstrated smooth and gummy meat surface due to the solubilization of myofibrillar proteins and the distortion of connective tissue in treated raw meats; and in the case of cooked meat, treatment caused the fragmentation of myofibrils, which might be responsible for a lower shear value in salt-bicarbonate treated beef biceps femoris muscle.

Apoptosis-Inducing Activity of HPLC Fraction from Voacanga globosa (Blanco) Merr. on the Human Colon Carcinoma Cell Line, HCT116

  • Acebedo, Alvin Resultay;Amor, Evangeline Cancio;Jacinto, Sonia Donaldo
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.2
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    • pp.617-622
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    • 2014
  • Voacanga globosa (Blanco), a plant endemic to the Philippines, is traditionally used especially by indigenous people of Bataan in the treatment of ulcers, wounds and tumorous growths. This study aimed to provide scientific evidence to therapeutic properties by determining cytotoxic and pro-apoptotic activity of HPLC fractions from leaves on HCT116 human colon carcinoma and A549 human lung carcinoma cell lines. Ethanolic extraction was performed on V globosa leaves followed by hexane and ethyl acetate partitioning. Silica gel column chromatography and high performance liquid chromatography (HPLC) produced MP1, MP2 and MP3 fractions. Cytotoxic activity of the fractions was determined through MTT assay against the cancer cell lines HCT116 and A549 and the non-cancer AA8 Chinese hamster ovarian cell line. Pro-apoptotic activities of the most active fractions were further assessed through DAPI staining, TUNEL assay and JC-1 mitochondrial membrane potential assay with HCT116 cells. While the MPI fraction exerted no significant activity against all cell lines tested, MP2 and MP3 fractions demonstrated high toxicity against HCT116 and A549 cells. The MP3 fraction induced formation of apoptotic bodies, condensed DNA and other morphological changes consistent with apoptosis of HCT116 cells and TUNEL assay showed significant increase in DNA fragmentation over time. In these cells, the MP3 fraction also induced mitochondrial membrane destabilization, which is generally associated with the beginning of apoptosis. Phytochemical analysis demonstrated the presence only of saponins and terpenoids in the MP3 fraction. The results indicate that the MP3 fraction exerts cytotoxic activity on HCT116 cells via induction of apoptosis triggered by loss of mitochondrial membrane potential crucial for cell survival.

In Vitro Antitumor Properties of an Isolate from Leaves of Cassia alata L

  • Olarte, Elizabeth Iglesias;Herrera, Annabelle Aliga;Villasenor, Irene Manese;Jacinto, Sonia Donaldo
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.5
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    • pp.3191-3196
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    • 2013
  • Leaf extracts of Cassia alata L (akapulko), traditionally used for treatment of a variety of diseases, were evaluated for their potential antitumor properties in vitro. MTT assays were used to examine the cytotoxic effects of crude extracts on five human cancer cell lines, namely MCF-7, derived from a breast carcinoma, SK-BR-3, another breast carcinoma, T24 a bladder carcinoma, Col 2, a colorectal carcinoma, and A549, a nonsmall cell lung adenocarcinoma. Hexane extracts showed remarkable cytotoxicity against MCF-7, T24, and Col 2 in a dose-dependent manner. This observation was confirmed by morphological investigation using light microscopy. Further bioassay-directed fractionation of the cytotoxic extract led to the isolation of a TLC-pure isolate labeled as f6l. Isolate f6l was further evaluated using MTT assay and morphological and biochemical investigations, which likewise showed selectivity to MCF-7, T24, and Col 2 cells with $IC_{50}$ values of 16, 17, and 17 ${\mu}g/ml$, respectively. Isolate f6l, however, showed no cytotoxicity towards the non-cancer Chinese hamster ovarian cell line (CHO-AA8). Cytochemical investigation using DAPI staining and biochemical investigation using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)-a method used to detect DNA fragmentation-together with caspase assay, demonstrated apoptotic cell death. Spectral characterization of isolate f6l revealed that it contained polyunsaturated fatty acid esters. Considering the cytotoxicity profile and its mode of action, f6l might represent a new promising compound with potential for development as an anticancer drug with low or no toxicity to non-cancer cells used in this study.

Effects of Temperature, Irradiance, and Nutrient Type on the Fragment Growth of Green Tide Alga Cladophora vadorum (녹조 대발생종 금발대마디말(Cladophora vadorum)의 절편 생장에 온도, 조도 및 영양염 종류가 미치는 영향)

  • Na, Yeon Ju;Jeon, Da Vine;Lee, Jung Rok;Kim, Young Sik;Choi, Han Gil;Nam, Ki Wan
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.49 no.5
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    • pp.657-664
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    • 2016
  • The green macroalga Cladophora vadorum bloomed along the coast at Sangrok Beach, Buan, South Korea, in September 2015. To elucidate the cause of bloom, the effects of environmental factors on the vegetative growth of adult fragments were examined. Growth experiments were carried out under different combinations of temperatures and irradiances, and with a single factor of nutrients (nitrogen, phosphorus). The maximal growth of C. vadorum was reported under the combination of 25°C and 100 μmol photons m−2s−1. The species grew under a wide range of N and P concentrations. The growth of C. vadorum peaked at 50 μM PO43−, 80 μM NH4+, and 100 μM NO3. Adult fragments formed holdfasts and new branches within 3 days in culture and became adults, showing polarized growth patterns, in 2 weeks. This is the first report showing the development of numerous bladelets from a segment in Cladophora species. The present results indicate that Cladophora blooms appear under growth conditions that are favorable in terms of temperatures, irradiance, and nutrients via fragment growth patterns producing rapid holdfasts and many bladelets.

A Study for Removing Road Shields from Mobile Mapping System of the Laser Data using RTF Filtering Techniques (RTF 필터링을 이용한 모바일매핑시스템 레이저 데이터의 도로 장애물 제거에 관한 연구)

  • Song, Hyun-Kun;Kang, Byoung-Ju;Lee, Sung-Hun;Choi, Yun-Soo
    • Journal of Korean Society for Geospatial Information Science
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    • v.20 no.1
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    • pp.3-12
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    • 2012
  • It is a global trend to give attention to generating precise 3D navigation maps since eco-friendly vehicles have become a critical issue due to environmental protection and depletion of fossil fuels. To date, Mobile Mapping System (MMS) has been a efficient method to acquire the data for generating the 3D navigation maps. To achieve this goal so far in the Mobile Mapping System using the data acquisition method has been proposed to be most effective. For this study the basic RTF filter algorithm was applied to modify to fit MMS quantitative analysis derived floor 99.71%, 99.95% of the highly non-producers to maintain accuracy and high-precision 3D road could create DEM. In addition, the roads that exist within the cars, roadside tree, road cars, such as the median strips have been removed to shields it takes to get results effectively, and effective in practical applications and can be expected to improve operational efficiency is considered.