• Title/Summary/Keyword: flow cytometry analysis

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The Study of anti-cancer mechanism with Cobrotoxin on Human prostatic cancer cell line(PC-3) (전립선 암세포에 대한 Cobrotoxin의 항암(抗癌) 기전(機轉) 연구(硏究))

  • Chae, Sang-jin;Song, Ho-seup
    • Journal of Acupuncture Research
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    • v.22 no.3
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    • pp.169-183
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    • 2005
  • Objective : The purpose of this study was to investigate the anti-caner effect of cobrotoxin on the prostatic cancer cell line (PC-3).The goal of study is to ascertain whether cobrotoxin inhibits tile cell growth and cell cycle of PC-3, or the expression of relative genes and whether the regression of PC-3 cell growth is due to cell death or the expression of gene related to apoptosis. Methods : After the treatment of Pc-3 cells with cobrotoxin, we performed 형광현미경, MTT assay, Western blotting, Flow cytometry, PAGE electrophoresis and Surface plasmon resonance analysis to identify the cell viability, cell death, apoptosis, the changes of cell cycle and the related protein, Adk, MAP kinase. Results : 1. Compared with normal cell, the inhibition of cell growth reduced in proportion with the dose of cobrotoxin(0-16nM) in PC-3. 2. Cell viabilities of 0.1, 1, 4nM cobrotoxin treatment were decreased and those of 8, 16nM were decreased significantly. 3. S phase of cell cycle was decreased at the group of 1, 2, 4, 8, 16nM cobrotoxin, but M phase was increased at 0.1, 1, 2, 4, 8, 16nM cobrotoxin. 4. Cox-2 expression after cobrotoxin was peaked at 12hours and was decreased significantly after 6, 12, 24 hours. 5. The expression of Cdk4 was decreased dose-dependently at 1, 2, 4, 8nM cobrotoxin and was decreased siginificantly at 4, 8nM Cyclin D1 was decreased at 1, 2, 4, 8nM and Cycline E was not changed. Cycline B was decreased at 1, 2, 4, 8nM dose-dependently and was decreased siginificanlty at 2, 4, 8nM. 6. The expression of Akt was decreased at 1, 2, 4, 8nM dose-dependently and was decreased significantly at 2, 4, 8nM. 7. ERK was increased at 1, 2nM and decreased at 4, 8nM, p-ERK was increased at 1, 2, 4 nM, but decreased at 8nM. JNK and p-JNK were increased at 1, 4, 8 nM. p38 was increased at 2nM p-p38 was increased at lnM but decreased significantly at 2, 4, 8nM. 8. The nucli of normal cells were stained round and homogenous in DAPI staining, but those of PC-3 were stained condense and splitted. Apoptosis was increased dose-dependently at 2, 4, 8, 16nM and increased significantly at 2, 4, 8, 16nM. 9. Bax wasn`t changed at 1, 2, 4, 8nM and Bcl-2 was decreased significantly at 1, 2, 4, 8nM. Caspase 3 and 9 weren`t changed at 1, 2, 4nM but were decreased significantly at 8nM. Conclusions : These results indicate that cobrotoxin inhibits the growth of prostate Cancer cells, has anti-cancer effects by inducing apoptosis.

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Phytoplankton Diversity and Community Structure Driven by the Dynamics of the Changjiang Diluted Water Plume Extension around the Ieodo Ocean Research Station in the Summer of 2020 (2020년 하계 장강 저염수가 이어도 해양과학기지 주변 해역의 식물플랑크톤 다양성 및 개체수 변화에 미치는 영향)

  • Kim, Jihoon;Choi, Dong Han;Lee, Ha Eun;Jeong, Jin-Yong;Jeong, Jongmin;Noh, Jae Hoon
    • Journal of the Korean Society of Marine Environment & Safety
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    • v.27 no.7
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    • pp.924-942
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    • 2021
  • The expansion of the Changjiang Diluted Water (CDW) plume during summer is known to be a major factor influencing phytoplankton diversity, community structure, and the regional marine environment of the northern East China Sea (ECS). The discharge of the CDW plume was very high in the summer of 2020, and cruise surveys and stationary monitoring were conducted to understand the dynamics of changes in environmental characteristics and the impact on phytoplankton diversity and community structure. A cruise survey was conducted from August 16 to 17, 2020, using R/V Eardo, and a stay survey at the Ieodo Ocean Research Station (IORS) from August 15 to 21, 2020, to analyze phytoplankton diversity and community structure. The southwestern part of the survey area exhibited low salinity and high chlorophyll a fluorescence under the influence of the CDW plume, whereas the southeastern part of the survey area presented high salinity and low chlorophyll a fluorescence under the influence of the Tsushima Warm Current (TWC). The total chlorophyll a concentrations of surface water samples from 12 sampling stations indicated that nano-phytoplankton (20-3 ㎛) and micro-phytoplankton (> 20 ㎛) were the dominant groups during the survey period. Only stations strongly influenced by the TWC presented approximately 50% of the biomass contributed by pico-phytoplankton (< 3 ㎛). The size distribution of phytoplankton in the surface water samples is related to nutrient supplies, and areas where high nutrient (nitrate) supplies were provided by the CDW plume displayed higher biomass contribution by micro-phytoplankton groups. A total of 45 genera of nano- and micro-phytoplankton groups were classified using morphological analysis. Among them, the dominant taxa were the diatoms Guinardia flaccida and Nitzschia spp. and the dinoflagellates Gonyaulax monacantha, Noctiluca scintillans, Gymnodinium spirale, Heterocapsa spp., Prorocentrum micans, and Tripos furca. The sampling stations affected by the TWC and low in nitrate concentrations presented high concentrations of photosynthetic pico-eukaryotes (PPE) and photosynthetic pico-prokaryotes (PPP). Most sampling stations had phosphate-limited conditions. Higher Synechococcus concentrations were enumerated for the sampling stations influenced by low-nutrient water of the TWC using flow cytometry. The NGS analysis revealed 29 clades of Synechococcus among PPP, and 11 clades displayed a dominance rate of 1% or more at least once in one sample. Clade II was the dominant group in the surface water, whereas various clades (Clades I, IV, etc.) were found to be the next dominant groups in the SCM layers. The Prochlorococcus group, belonging to the PPP, observed in the warm water region, presented a high-light-adapted ecotype and did not appear in the northern part of the survey region. PPE analysis resulted in 163 operational taxonomic units (OTUs), indicating very high diversity. Among them, 11 major taxa showed dominant OTUs with more than 5% in at least one sample, while Amphidinium testudo was the dominant taxon in the surface water in the low-salinity region affected by the CDW plume, and the chlorophyta was dominant in the SCM layer. In the warm water region affected by the TWC, various groups of haptophytes were dominant. Observations from the IORS also presented similar results to the cruise survey results for biomass, size distribution, and diversity of phytoplankton. The results revealed the various dynamic responses of phytoplankton influenced by the CDW plume. By comparing the results from the IORS and research cruise studies, the study confirmed that the IORS is an important observational station to monitor the dynamic impact of the CDW plume. In future research, it is necessary to establish an effective use of IORS in preparation for changes in the ECS summer environment and ecosystem due to climate change.