• Title/Summary/Keyword: sub-G1 DNA

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Induced apoptosis in human Uterine Leiomyoma Cells by treatment with Chiljehyangbu-hwan (칠제향부환(七製香附丸)이 자궁근종세포의 성장억제와 세포자멸사에 미치는 영향)

  • Kim, Seuk-Jung;Beak, Seung-Hee;Kim, En-Ha;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.20 no.2
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    • pp.25-42
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    • 2007
  • Purpose : Uterine leiomyoma (fibroids) are benign smooth muscle tumors originating from the myometrium. These benign neoplasms of monoclonal origin are typically diagnosed during the reproductive years, occurring only after puberty and tending to regress after menopause. In the present study we used Chiljehyangbu-hwan to determine its growth inhibitory effect and apoptosis in human uterine leiomyoma cells. Methods : Primary cultured human uterine leiomyoma cells were treated with Chiljehyangbu-hwan. Cell viability analysis was analyzed by MTS assay and FACS was performed to ascertain the effects Chiljehyangbu-hwan. DNA fragmentation analysis and casapase-3 activity test were done. Expression of apoptosis related proteins were evaluated by Western blot analysis. Results : Cell viability was significantly influenced by Chiljehyangbu-hwan treatment in a dose-dependent manner in leiomyoma cells compare to normal myometrial cells. FACS showed that Chiljehyangbu-hwan induced Sub G1 arrest. DNA fragmentation assay was carried out and apoptosis was detected. Activation of caspase-3, down-regulation of Bcl-2, with concomitant increased expression in Bid and Bax were observed. Chiljehyangbu-hwan treatment of uterine leiomyoma cells resulted in a concentration-dependent cell death induced via the mitochondrial pathway.

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Growth Arrest and Apoptosis of Human Uterine Cervical Carcinoma Cells Induced by Trichosanthes Semen Extract (과루인이 자궁경부암세포의 성장억제 및 세포고사에 미치는 영향)

  • Lee Jeong Gu;Kim Yeon Hee;Lee Dong Nyung;Kim Hyung Jun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.4
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    • pp.965-972
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    • 2005
  • To investigate the effects of Trichosanthes semen extract on the growth and apoptosis of human uterine cervical carcinoma cells. Effects of Trichosanthes semen extract on the growth of ME-180 cells were assayed by MTT assay. Apoptosis induced by Trichosanthes semen extract was detected by fluorescent microscopy, DNA fragmentation analysis and flow cytometry. Caspase-3 and caspase-8 activities were assayed. Trichosanthes semen extract induced ME-180 cells to die in a dose- and time-dependent manner. ME-180 cells treated with Trichosanthes semen extract exhibited typical characteristics of apoptosis. The population of Sub-G1 cells increased significantly, and the cells represented the reduced size, condensed chromatin and apoptotic bodies. They showed the decreased mitochondrial membrane potential and increased activities of caspase-3 and caspase-8. The results suggest that Trichosanthes semen extract induced ME-180 cell apoptosis and the activation of caspase and mitochondrial pathway were involved in the process of Trichosanthes semen extract-induced apoptosis.

Protective Effects of a Herbal Composition (HemoHIM) Against Apoptosis Induced by Oxidative Stress of Hydrogen Peroxide (과산화수소의 산화적 스트레스로 유도된 Apoptosis에 대한 생약복합조성물(HemoHIM)의 방호효과 평가)

  • Shin, Sung-Hae;Kim, Do-Soon;Kim, Mi-Jung;Kim, Sung-Ho;Jo, Sung-Kee;Byun, Mung-Woo;Yee, Sung-Tae
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.9
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    • pp.1127-1132
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    • 2006
  • In our previous study, a novel herb mixture (HIM-I) of Angelica gigas radix, Cnidium officinale rhizoma, and Paeonia japonica radix was developed to protect the intestinal and immune systems and promote its recovery against radiation damage. A new herbal composition (HemoHIM) with the high immune modulating activity was developed from HIM-I. HIM-I was fractionated into ethanol fraction (HIM-I-E) and polysaccharide fraction (HIM-I-P). And HemoHIM was prepared by adding HIM-I-P to HIM-I. HemoHIM showed more effective than HIM-I in immune modulation as well as radioprotection. The present study is designed to investigate the protective effects of HIM-I, HIM-I-P, and HemoHIM on hydrogen peroxide $(H_2O_2)$ induced apoptosis of human promyelocytic leukemia (HL-60) cells. It was shown that $H_2O_2$ treatment reduced the viability of cells, and increased appearance of DNA ladders, hypodiploid (subG1) cells, and phosphatidylserine translocation level. Pretreatment of HemoHIM significantly reduced the cytotoxic effect induced by $H_2O_2$, associated with reducing the translocation of phosphatidylserine, hypodiploid cells and DNA ladders. HemoHIM appeared to be more protective than HIM-I against $H_2O_2$ induced apoptosis whereas, it exhibited similar activity to HIM-I-P. These results indicated that HemoHIM might be an useful agent for protection against oxidative stress $(H_2O_2)-induced$ apoptosis as well as immune modulation, especially since it is a relatively nontoxic natural product.

The effect of Sparganii Rhizoma on the proliferation inhibition of human uterine leiomyoma cell and expression of gene related cell apoptosis (삼릉(三稜)이 자궁근종세포의 증식억제와 세포자멸사 관련 발현에 미치는 영향)

  • Park, Chang-Gun;Baek, Seung-Hee;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.19 no.2
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    • pp.199-213
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    • 2006
  • Purpose : This study was aimed to investigate the inhibitory effect of Sparganii Rhizoma on the proliferation of human uterine leiomyoma cells and the expression of gene related the mechanism of cell apoptosis. Methods : This study was evaluated the number of death cells treated with indicated concentration of Sparganii Rhizoma and investigated cell death rate by MTS assay. Furthermore, fluorescence-activated cell sorter analysis and DNA fragmentation assay were used to dissect between necrosis and apoptosis. and then we observed the differential gene expression by western blot analysis. Results :1) The inhibitory effect on the growth of uterine leiomyoma cell treated with Sparganii Rhizoma was increased in a dose dependent manner. 2) As the result of FACS analysis, subG1 phase incrase was observed 23.49% inuterine leiomyoma cell treated with Sparganii Rhizoma at $500\;{\mu}g/ml$ compared to control.. 3) The gene expression of p53, p21 related cell apoptosis was increased according to increasing concentration but p27 was none exchanged. 4) The expression of cyclin A, D and E was decreased in a concentration proportional and then the dephosphorylation of pRb was increased. 5) The character of apoptosis, DNA fragmentation was significantly observed according to increasing concentration. 6) The expression of pro-caspase3 were decreased dependent on treatment concentration and activated PARP took place. Conclusion : The inhibitory effect of Sparganii Rhizoma on the proliferation of human uterine leiomyoma cells was observed with apoptosis and cell cycle arrest. These data suggest that Sparganii Rhizoma might be candidate of medical therapy for uterine leiomyoma.

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Cultivated Orostachys japonicus Induces Apoptosis in Human Colon Cancer Cells (인체 대장암 세포주 SW480에서 재배 와송의 세포 사멸 유도 효과)

  • Kim, Jae-Yong;Jung, Eun-Jung;Won, Yeong-Seon;Lee, Ju-Hye;Shin, Dong-Young;Seo, Kwon-Il
    • Korean Journal of Food Science and Technology
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    • v.44 no.3
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    • pp.317-323
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    • 2012
  • This study was performed to elucidate the anticancer activities and the mechanism of chloroform fractions from cultivated Orostachys japonicus (CFCOJ) in human colon cancer cells. CFCOJ markedly decreased viable cell numbers in both a dose-dependent and time-dependent manner within SW480 cells. Cell death induced by CFCOJ increased cell populations in the sub-G1 phase, as well as the formation of apoptotic bodies, nuclear condensation, and induced DNA fragmentation. CFCOJ-induced apoptosis was associated with the activation of initiator caspase-8 and -9, as well as the effector caspase-3. CFCOJ also stimulated Bid cleavage, indicating that the apoptotic action of caspase-8-mediated Bid cleavage leads to the activation of caspase-9. CFCOJ increased the expression of the proapoptotic protein, Bax, and decreased the expression of the antiapoptotic protein, Bcl-2. These results indicate that CFCOJ exert anticancer effects on human colon cancer SW480 cells through a caspase-dependent apoptotic pathway.

Antitumoral Compound , MCH-201 , an Effector on Proliferation and Morphology of Human Breast Tumor Cell Line, MCF-7 (인체유암세포주 MCF-7의 형태변화와 증식에 영향을 주는 항암활성물질, MCH-201)

  • Kim, Hang-Sub;Kim, Se-Eun;Kim, Young-Ho;Lee, Sung-Woo;Oh, Goo-Taeg;Kim, Hwan-Mook;Lee, Jung-Joon
    • Microbiology and Biotechnology Letters
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    • v.21 no.4
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    • pp.316-321
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    • 1993
  • MCH-201 was isolated from the mycelium of Streptomyces sp. Ba16 as a potent effector on proliferation and morphology of human breast tumor cell line, MCF-7. Morphological change could be observed at concentration between 2.5${\mu}$g/ml and 250pg/ml and showed cytotoxic effect at the concentration of more than 5${\mu}$g/ml. This compound also showed inhibitory effect on DNA synthesis of hepatoma cells, Hepa 1c1c7, and strong cytotoxic effect on proliferation of human tumor cell lines, A549 and XF498.

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Apoptosis Induction of HCT-15 Cells by Extracts of Undaria pinnatifida with Fermented Micro-organism (미역 발효추출물의 HCT-15 대장암 세포 사멸 유도 효과)

  • Kim, Tae-Yun;Han, Hyo-Sang;Lee, Young-Jong
    • The Korea Journal of Herbology
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    • v.28 no.4
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    • pp.33-40
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    • 2013
  • Objectives : To study the apoptosis effects of fermented Undaria pinnatifida extracts(FUP) against HCT-15 colon cancer cells. Method : By measuring cell proliferation, DNA fragmentation, cell cycle, morphology, and western blot from FUP, the study investigated the effects of the extractions had upon the HCT-15 colon cancer cells, and concluded that the inhibiting effects upon cells were induced by apoptosis. Result : FUP effectively inhibited the growth of HCT-15 colon cancer cells. After analyzing the DNA fragmentation, the study observed a DNA ladder, while examining the cells, and found an increase of sub-G1 hypodiploid cells. On the changes regarding the nucleus of the cells, a condensation of cells and chromatin, as well as an apoptotic body was clearly observed. By observing through western blot from FUP, the study found a decreased level of Bcl-2 from HCT-15 colon cancer cells, but the increased level of Bax and cleaved caspase-3, which as a result induced apoptosis, inhibiting the growth of HCT-15 colon cancer cells. FUP increased the natural death of HCT-15 colon cancer cells by the induction of apoptosis. FUP seemed to have no suppressing effect upon HL-60/MX2 cells. However, compared to the fucoidan, the study was able to clearly observe morphological changes of HCT-15 cells apoptosis, in a 1/2 concentration. Conclusion : FUP had antiproliferative effects on different kinds of cancer cells, while proving especially efficacious against colon cancer cells.

The Endoplasmic Reticulum Stress Response Mediates Shikonin-Induced Apoptosis of 5-Fluorouracil-Resistant Colorectal Cancer Cells

  • Piao, Mei Jing;Han, Xia;Kang, Kyoung Ah;Fernando, Pincha Devage Sameera Madushan;Herath, Herath Mudiyanselage Udari Lakmini;Hyun, Jin Won
    • Biomolecules & Therapeutics
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    • v.30 no.3
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    • pp.265-273
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    • 2022
  • Resistance to chemotherapeutic drugs is a significant problem in the treatment of colorectal cancer, resulting in low response rates and decreased survival. Recent studies have shown that shikonin, a naphthoquinone derivative, promotes apoptosis in colon cancer cells and cisplatin-resistant ovarian cells, raising the possibility that this compound may be effective in drug-resistant colorectal cancer. The aim of this study was to characterize the molecular mechanisms underpinning shikonin-induced apoptosis, with a focus on endoplasmic reticulum (ER) stress, in a 5-fluorouracil-resistant colorectal cancer cell line, SNU-C5/5-FUR. Our results showed that shikonin significantly increased the proportion of sub-G1 cells and DNA fragmentation and that shikonin-induced apoptosis is mediated by mitochondrial Ca2+ accumulation. Shikonin treatment also increased the expression of ER-related proteins, such as glucose regulatory protein 78 (GRP78), phospho-protein kinase RNA-like ER kinase (PERK), phospho-eukaryotic initiation factor 2 (eIF2α), phospho-phosphoinositol-requiring protein-1 (IRE1), spliced X-box-binding protein-1 (XBP-1), cleaved caspase-12, and C/EBP-homologous protein (CHOP). In addition, siRNA-mediated knockdown of CHOP attenuated shikonin-induced apoptosis, as did the ER stress inhibitor TUDCA. These data suggest that ER stress is a key factor mediating the cytotoxic effect of shikonin in SNU-C5/5-FUR cells. Our findings provide an evidence for a mechanism in which ER stress leads to apoptosis in shikonin-treated SNU-C5/5-FUR cells. Our study provides evidence to support further investigations on shikonin as a therapeutic option for 5-fluorouracil-resistant colorectal cancer.

Antioxidative activities of Artemisia capillaris-Fermented Hericium erinaceum Mycelium (인진쑥 노루궁뎅이 버섯균사체 발효물의 항산화 활성)

  • Kim, Seung-Sub;Kyeong, Inn-Goo;Lee, Mi-La;Kim, Dong-Goo;Shin, Ji-Young;Yang, Jin-Yi;Lee, Gwang-Ho;Eum, Won-Sik;Kang, Jung-Hoon
    • Journal of the Korean Applied Science and Technology
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    • v.31 no.4
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    • pp.719-730
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    • 2014
  • The hot water extract from Artemisia capillaris fermented with Hericium erinaceum mycelium (AC-HE) were assessed for the protection against oxidative modification of biological macromolecules and cell death. Antioxidant activity of AC-HE evaluated using 2,2-diphenyl-1-picrylhydrazyl radical, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical and peroxyl radical scavenging assays. AC-HE showed 61.73% DPPH radical scavenging activity at $500{\mu}g/mL$, 97.39% ABTS radical scavenging activity at $250{\mu}g/mL$, and 44.18% peroxyl radical scavenging activity at $100{\mu}g/mL$. AC-HE were shown to significantly inhibited DNA strand breakage induced by peroxyl radical. AC-HE also prevented peroxyl radical-mediated human serum albumin modification. AC-HE effectively inhibited $H_2O_2$ induced cell death and significantly increased of the 11.47% cell survival at $100{\mu}g/mL$. AC-HE also decreased intracellular reactive oxygen species (ROS) levels in $H_2O_2$-treated cells. The results suggested that AC-HE can contribute to antioxidant and protected cells from oxidative stress-induced cell injury.

Evaluation of Genetic Differentiation of Albizia lucida Populations from Eastern Region of the Indian Sub-continent by ISSR Markers

  • Aparajita, Subhashree;Rout, G.R.
    • Journal of Forest and Environmental Science
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    • v.24 no.1
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    • pp.27-34
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    • 2008
  • Level and distribution of genetic diversity in seven populations of Albizia lucida Benth. in eastern region of the Indian sub-continent were estimated using ISSR markers. Relatively higher level of genetic diversity within populations was observed in seven populations of A. lucida (mean of 0.38). From the result of AMOVA, majority of genetic diversity was allocated within populations (96.2%) resulting in a moderate degree of population differentiation. The observed distribution pattern of I-SSR variant among the populations was coincided with the typical pattern of long-lived woody tree species. Genetic relationships among the populations, reconstructed by UPGMA method, revealed two genetic groups. The population of Anugul and Bargarh turned out to be the most closely related despite a distance location between them. These formations will be of great value in the development of conservation plans for species exhibiting high levels of genetic differentiation due to fragmentation, such as indication of conservation unit size, which populations should be chosen as priority in conservation plans and which samples should be introduced in areas with a low number of individuals of A. lucida.

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