• Title/Summary/Keyword: cell fractions

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Effect of Acer tegmentosum M. Extracts on Hepatocarcinoma Cell (산겨릅나무 추출물의 간암세포의 증식억제 효과)

  • Kwon, Ha-Na;Bang, Woo-Suk;Kim, Joo-Young;Park, Jyung-Rewng;Jeon, Jeong-Ryae
    • Korean Journal of Food Science and Technology
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    • v.43 no.6
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    • pp.787-790
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    • 2011
  • The objective of this study was to investigate the anticancer effects of Acer tegmentosum M. extracts. HepG2 hepatocarcinoma cells were treated with ethanol, chloroform, ethylacetate, butanol, aqueous fraction and hot water extract. The antiproliferative effect was evaluated by trypan blue exclusion, MTT-based viability assay and morphology. The trypan blue test showed that anticancer effect of the A. tegmentosum M. extracts on HepG2 cells increased gradually in proportion to the increasing concentration of the fractions. The butanol fraction showed the highest anticancer activity against HepG2 cells (p<0.05). The MTT assay indicated that the growth inhibition by the butanol fraction was dose-dependent. These results suggest that A. tegmentosum M. has the potential to inhibit the growth of hepatocarcinoma cells.

Accumulation of Crude Lipids, Phenolic Compounds and Iron in Rusty Ginseng Root Epidermis (적변삼 외피에서 지질, 페놀성물질 및 철 성분의 축적에 관한 연구)

  • Lee, Tae-Su;Mok, Sung-Kyun;Cheon, Seong-Ki;Yoon, Jong-Hyuk;Baek, Nam-In;Choe, Jyung
    • Journal of Ginseng Research
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    • v.28 no.3
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    • pp.157-164
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    • 2004
  • The study was conducted to investigate the tissue and chemical characteristics of rusty root epidermal cells. In histological study, the rusty symptoms were frequently observed in the epidermis of ginseng root and to be yellow under microscopic observation. Disks of the epidermal cell tissue of the rusty root were usually 2 and 3 times greater in the number of cell layer and thickness of cell wall than the healthy root, respectively. The color degree of methanol extracts from the rusty root epidermis was 5.5 times higher than that of the healthy root. And the extracts of rust matter in the root epidermis were easily dissolved in polar solvents compared to nonpolar solvents. UV-absorption spectra of methanol extracts in various fractions of phenolics showed a maximum peak between 275∼280 nm. The crude lipids and phenolic compounds such as acid insoluble bound phenolics, acid insoluble esterified phenolics, acid insoluble condensed phenolics, insoluble bound phenolics and free phenolics were also more in the rusty root epidermis than in the healthy one. Fe content in the rusty root epidermis was 2.7 times higher than that of healthy one. It was presumed that the phenolic compounds(precursor of the rusty) in association with lipid and iron in the root epidermis might defence the root when ginseng root was depressed by the unfavorable conditions in soil and/or portions of a root system were subjected to anoxic conditions.

Anti-proliferative and Pro-apoptotic Effects by Lees Extracts of Ehwa Makgeolli Containing Oriental Herbs (한방이화주 주박 추출물에 의한 암세포 항성장 및 세포사멸 기전 연구)

  • Kwon, Min-Jeong;Lee, Seung Hoon;Chung, Chung Wook;Sohn, Ho-Yong;Shin, Woo-Chang;Kim, Jong-Sik
    • Journal of Life Science
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    • v.26 no.2
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    • pp.241-246
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    • 2016
  • In the present study, ethanol extracts and their subsequent organic solvent fractions were extracted from the lees of Ehwa Makgeolli containing oriental herbs, a commercialized traditional Korean rice wine, and the prepared lees samples were designated as from KSD-E3-1 to KSD-E3-5. First, their effects on cell viability and on the expression of pro-apoptotic ATF3 and NAG-1 genes in human colorectal HCT116 cells were investigated. Among the treated lees samples, the hexane fraction (KSD-E3-2) and the ethyl acetate fraction (KSD-E3-3) of lees extracts from Ehwa Makgeolli significantly reduced cell viabilities, in a dose dependent manner. The treatment with KSD-E3-2 and KSD-E3-3 also increased the expression of pro-apoptotic NAG-1 and ATF-3 genes and their proteins, which were detected with RT-PCR and Western blot analysis, respectively. In addition, poly-(ADP-ribose) polymerase (PARP) cleavage was detected by treatment with the fraction KSD-E3-3, indicating that KSD-E3-3 could induce apoptosis in HCT116 cells. Interestingly, this PARP cleavage was recovered by transfection of NAG-1 small interfering RNA. The results indicate that NAG-1 is one of the genes responsible for apoptosis induced by the fraction KSD-E3-3 from Ehwa Makgeolli. Overall, the findings may help in understanding the molecular mechanisms of the anti-proliferative and pro-apoptotic activities mediated by the lees of Ehwa Makgeolli.

Induction of Apoptosis by Treatment of Human Prostate Cancer LNCaP Cells with Methanol Fractions from Prunus mume (매실(Prunus mume) 메탄올 분획물의 처리에 따른 인체 전립선암세포 LNCaP의 apoptosis 유도 효과)

  • Kim, Hwi-gon;Kim, Jeong-Ho;Heo, Ji-An;Won, Yeong-Seon;Seo, Kwon-Il
    • Journal of Life Science
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    • v.31 no.3
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    • pp.321-329
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    • 2021
  • This study examined the growth inhibitory effect of the methanol fraction of maesil (Prunus mume) extract (MMF) on LNCaP, PC-3, and RC-58T human prostate cancer cell lines. Among these cell lines, LNCaP was the most sensitive to the inhibitory effects of MMF. Observation of the morphology and apoptotic body formation in the LNCaP cells revealed morphological changes, nuclear damage, and condensation in response to MMF treatment. The suppressive effect of MMF was related to the intrinsic apoptosis pathway, as indicated by increased expression of the pro-apoptotic proteins Bax, capase-3, capase-9, and PARP and decreased expression of the anti-apoptotic protein Bcl-2. Combined treatment with MMF and the AIF inhibitor N-phenylmalemide (N-PM) indicated that MMF treatment alone had a significant growth suppression effect. The involvement of the extrinsic apoptosis pathway was also confirmed by increased expression of AIF and Endo G. The growth suppression effect of MMF was also significant when compared to the effects of a combination of the PI3K inhibitor LY294002 and MMF. The reduced expression of PI3K, p-Akt, and p-mTOR confirmed the involvement of the PI3K/Akt/ mTOR signaling pathway in regulating the anti-proliferative properties of MMF. In conclusion, the growth suppression effect of MMF in the LNCaP human prostate cancer cell line shows the possibility of using this natural product in functional foods.

Correlation Between the Parameters of Radiosensitivity in Human Cancer Cell Lines (인체 암세포주에서 방사선감수성의 지표간의 상호관계)

  • Park, Woo-Yoon;Kim, Won-Dong;Min, Kyung-Soo
    • Radiation Oncology Journal
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    • v.16 no.2
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    • pp.99-106
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    • 1998
  • Purpose : We conducted clonogenic assay using human cancer cell lines (MKN-45, PC-14, Y-79, HeLa) to investigate a correlation between the parameters of radiosensitivity. Materials and Methods : Human cancer cell lines were irradiated with single doses of 1, 2, 3, 5, 7 and 10Gy for the study of radiosensitivity and subrethal damage repair capacity was assessed with two fractions of 5Gy separated with a time interval of 0, 1, 2, 3, 4, 6 and 24 hours. Surviving fraction was assessed with clonogenic assay using $Sperman-H\"{a}rbor$ method and mathematical analysis of survival curves was done with linear-quadratic (LQ) , multitarget-single hit(MS) model and mean inactivation dose$(\v{D})$. Results : Surviving fractions at 2Gy(SF2) were variable among the cell lines, ranged from 0.174 to 0.85 The SF2 of Y-79 was lowest and that of PC-14 was highest(p<0.05, t-test). LQ model analysis showed that the values of $\alpha$ for Y-79, MKN-45, HeLa and PC-14 were 0.603, 0.356, 0.275 and 0.102 respectively, and those of $\beta$ were 0.005, 0.016, 0.025 and 0.027 respectively. Fitting to MS model showed that the values of Do for Y-79. MKN-45, HeLa and PC-14 were 1.59. 1.84. 1.88 and 2.52 respectively, and those of n were 0.97, 1.46, 1.52 and 1 69 respectively. The $\v{D}s$ calculated by Gauss-Laguerre method were 1.62, 2.37, 2,01 and 3.95 respectively So the SF2 was significantly correlated with $\alpha$, Do and $\v{D}$. Their Pearson correlation coefficiencics were -0.953 and 0,993. 0.999 respectively(p<0.05). Sublethal damage repair was saturated around 4 hours and recovery ratios (RR) at plateau phase ranged from 2 to 3.79. But RR was not correlated with SF2, ${\alpha}$, ${\beta}$, Do, $\v{D}$. Conclusion : The intrinsic radiosensitivity was very different among the tested human cell lines. Y-79 was the most sensitive and PC-l4 was the least sensitive. SF2 was well correlated with ${\alpha}$, Do, and $\v{D}$. RR was high for MKN-45 and HeLa but had nothing to do with radiosensitivity parameters. These basic parameters can be used as baseline data for various in vitro radiobiological experiments.

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Effect of Cytarabine, Melphalan, and Total Body Irradiation as Conditioning for Autologous Stem Cell Transplantation for Patients with AML in First Remission (1차 관해된 급성 골수성 백혈병에서 자가 조혈모세포 이식을 위한 Cytarabine, Melphalan, 전신 방사선치료의 효과)

  • Kang Ki Mun;Choi Byung Ock;Chai Gyu Young;Kang Young Nam;Jang Hong Sek;Kim Hee Jae;Min Wo Sung;Kim Chun Choo;Choi Ihl Bohng
    • Radiation Oncology Journal
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    • v.21 no.3
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    • pp.192-198
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    • 2003
  • Purpose: Current results of autologous stem cell transplantation (SCT) suggest that this procedure may prolong disease free survival In patients with acute myeloid leukemia (AML). Autologous SCT is increasingly used as treatment for AML in first remission. The aim of this study was to evaluate the outcome of autologous SCT for patients with AML in first remission treated by autologous SCT using cytarabine, melphalan and total body irradiation (TBI) as the conditioning regimen. Materials and Methods: Between January 1995 and December 1999, 29 patients with AML in first remission underwent autologous SCT. The median age of patients was 33 years (range, 16 to 47). The conditioning regimen consisted of cytarabine ($3.0\;gm/m^2$ for 3 days), melphalan ($100\;gm/m^2$ for 1 day) and TBI (total 1000 cGy in five fractions over 3 days). Results: The median follow up was 40 months with a range of 3 to 58 months. The 4-year cumulative probability of disease free survival was 69.0%, and median survival was 41.5 months. The 4-year relapse rate was 27.6%. The factor Influencing disease free survival and relapse rate was the French-American-British (FAB) classification ($M_3$ group vs. other groups; p=0.048, p=0.043). One patient died from treatment-related toxicity. Conclusion:: Although the small number of patients does not allow us to draw any firm conclusion, our results were encouraging and suggest that the association of cytarabine, melphalan and TBI as a conditioning regimen for autologous SCT for AML on first remission appears to be safe and effective.

The Use of MTT Assay, In Vitro and Ex Vivo, to Predict the Radiosensitivity of Colorectal Cancer (In-vitro와 Ex-vivo MTT Assay를 통한 직장암의 방사선치료 감수성 예측 가능성 검증)

  • Kim, Ji-Eun;Kim, Mi-Sook;Kang, Chang-Mo;Kim, Jong-Il;Shin, Hye-Kyung;Choi, Chul-Won;Seo, Young-Seok;Ji, Young-Hoon
    • Radiation Oncology Journal
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    • v.26 no.3
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    • pp.166-172
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    • 2008
  • Purpose: The measurement of radiosensitivity of individuals is useful in radiation therapy. Unfortunately, the measurement of radiation survival using a clonogenic assay, which is the established standard, can be difficult and time consuming. The aim of this study is to compare radiosensitivity results obtained from the MTT and clonogenic assays, and to evaluate whether the MTT assay can be used on clinical specimens. Materials and Methods: HCT-8, LoVo, CT-26, and WiDr were the colon cancer cell lines used for this study. The clonogenic assay was performed to obtain the cell survival curves and surviving fractions at a dose of 2 Gy ($SF_2$) as the standard technique for radiosensitivity. Also, the MTT assay was performed for each of the cell lines (in vitro). To simulate clinical specimens, the cell lines were inoculated into nude mice, removed when the tumors reached 1 cm in diameter, and chopped. Next, the tumors were subjected to the same process involved with the MTT assay in vitro. The inhibition rates (IR) of 10 Gy or 20 Gy of irradiation for in vitro and ex vivo were calculated based on the optical density of the MTT assay, respectively. Results: According to $SF_2$ and the cell survival curve, the HCT-8 and WiDr cell lines were more resistant to radiation than LoVo and CT-26 (p<0.05). The IR was measured by in vitro. The MTT assay IR was 17.3%, 21%, 30% and 56.5% for the WiDr, HCT-8, LoVo and CT-26 cell lines, respectively. In addition, the IR measured ex vivo by the MTT assay was 23.5%, 26%, 38% and 53% in the HCT-8, WiDr, LoVo and CT-26 tumors, respectively. Conclusion: The radiosensitivity measured by the MTT assay was correlated with the measures obtained from the clonogenic assay. This result highlights the possibility that the MTT assay could be used in clinical specimens for individual radiosensitivity assays.

Discontinuous Percoll Gradients Enrich X-Bearing Porcine Sperms and Female Embryos (불연속 Percoll 원심분리에 의한 돼지 X-정자와 자성배아에 관한 연구)

  • Shim, Dae-Yong;Yoo, Seong-Jin;Kang, Han-Seung;Yoo, Jeong-Min;Lee, Chae-Kwan;Kang, Sung-Goo
    • Development and Reproduction
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    • v.5 no.1
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    • pp.47-52
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    • 2001
  • Predetermination of sex in livestock of offpring is in great demand and is of critical importance to providing for the most efficient production of the animal ariculture. Such a sexing techlology would also enhance the economy of conventional artificial insemination(AI) and aid the porcine industry. The purpose of this study was to evaluate the efficiency of enriching X-bearing porcine sperm using discontinuous percoll gradients and PCR mefhod. Semen was collected from mature boars of proven fertility center (AI center KimHae). Sperm was leaded on the isotonic discontinuous percoll gradient and then it was centrifuged at 120 ${\times}$ g for 20 minutes. After centrifugation, sperm included in each fraction were recovered (7${\times}$10$^6$ sperms/ml) and then sperm genomic DNA was extractedfor the PCR. SRY gene was used to evaluate the ratio between X and Y sperm in the separated fractions. Ju viro ffrtilization wascarried out by adding the unseparated sperm (control) or separated (experimental poop) to the matured oocytes in TCM-199. Embryos for sex determination were obtained at 2 cell stage and then was used for SRY gene amplification. After centrifugation of discontinuous percoll gradient, the most motile sperm was obtained at 95% fiaction (94.4% ${\pm}$ 5.1%, p < 0.01). The PCR analysis evaluated that 30%, 50% and 65% fractions were Y sperm rich, whereas 80% and 95% fractions were X sperm rich. PCR analysis with each porcineembryo showed that 33.3% of control and 66.7% of experimental group were determined as female embryos. In conclusion, in vitro matured oocytes inseminated with sperms (95% fraction) prepared by percoll gradient centrifugation showed high fertilization rates and female embryos than control sperms.

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Antioxidant and Cellular Protective Effects of Moringa oleifera Leaves Extract (드럼스틱 잎 추출물의 항산화 및 세포보호 효과)

  • Xuan, Song Hua;Kim, A Rang;Jeong, Yoon Ju;Lee, Nan Hee;Park, Soo Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.42 no.3
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    • pp.217-226
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    • 2016
  • In this study, we investigated the antioxidative and cellular protective effects on HaCaT cells and erythrocytes of Moringa oleifera (M. oleifera) leaves extract and its fractions. All experiments were performed with 50% ethanol extract, ethyl acetate fraction and aglycone fraction of M. oleifera leaves. The free radical scavenging activity ($FSC_{50}$) of the extract and fractions of M. oleifera leaves were in the following order: 50% ethanol extract ($77.10{\mu}g/mL$) < ethyl acetate fraction ($20.63{\mu}g/mL$) < aglycone fraction ($17.00{\mu}g/mL$) by using the 1, 1-diphenyl-2-picrylhydrazyl. In $Fe^{3+}-EDTA/H_2O_2$ system using the luminol, reactive oxygen species (ROS) scavenging activities (total antioxidant capacity, $OSC_{50}$) of aglycone fraction ($OSC_{50}=0.63{\mu}g/mL$) was the strongest among all extracts, which was much higher than L-ascorbic acid ($1.50{\mu}g/mL$). In the $^1O_2$-induced cellular damage of erythrocytes, the cellular protective effects of 50% ethanol extract (${\tau}_{50}=46.9min$) and aglycone fraction (${\tau}_{50}=122.1min$) were higher than (+)-${\alpha}$-tocopherol (${\tau}_{50}=37.7min$), known as a lipophilic antioxidant at $10{\mu}g/mL$. After cell damage induced by $400mJ/cm^2$ UVB irradiation, the cellular protective effects of ethyl acetate and aglycone fraction of M. oleifera leaves extract were showed on the concentration from 0.20 to $1.56{\mu}g/mL$. These results suggest that M. oleifera leaves extract and its fractions can function as a natural antioxidant agent in cosmetics on skin exposed to UV radiation by protecting cellular membrane against ROS.

Antioxidant and Anti-inflammatory Effects of Ficus erecta var. sieboldii Leaf Extract in Murine Macrophage RAW 264.7 Cells (좁은잎천선과나무(Ficus erecta var. sieboldii) 잎 추출물이 대식세포 RAW 264.7 세포에서 미치는 항산화 및 항염증 효과)

  • Jung, Yong-Hwan;Ham, Young-Min;Yoon, Seon-A;Oh, Dae-Ju;Kim, Chang-Suk;Yoon, Weon-Jong
    • Korean Journal of Plant Resources
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    • v.31 no.4
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    • pp.303-311
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    • 2018
  • In this study, a preliminary evaluation of the antioxidant and anti-inflammatory activity of the Ficus erecta var. sieboldii (Miq.) King (FES) leaf extract has been performed to assess its potential as a natural resource for food and medicinal materials. FES was extracted using 70% EtOH and then fractionated sequentially using n-hexane, $CH_2Cl_2$, EtOAc, and n-BuOH. To screen for antioxidant and anti-inflammatory agents effectively, the inhibitory effect of the FES extracts on the production of oxidant stresses (DPPH, xanthine oxidase, and superoxide) and pro-inflammatory factors (NO, iNOS, COX-2, $PGE_2$, IL-6, and $IL-1{\beta}$) in the murine macrophage cell line RAW 264.7 activated with lipopolysaccharide (LPS) was examined. Among the sequential solvent fractions of FES, the $CH_2Cl_2$ and EtOAc fractions showed decreased production of oxidant stresses (DPPH, xanthine oxidase and superoxide), and the hexane and $CH_2Cl_2$ fractions of FES inhibited the production of pro-inflammatory factors (NO, iNOS, COX-2, and $PGE_2$). The $CH_2Cl_2$ fraction also inhibited the production of pro-inflammatory cytokines ($TNF-{\alpha}$, IL-6, and $IL-1{\beta}$). These results suggest that FES has a significant effects on the production of oxidant stresses and pro-inflammatory factors and may be used a natural resource for antioxidant and anti-inflammatory agents.