• 제목/요약/키워드: Cell fractionation

검색결과 248건 처리시간 0.035초

돼지감자로부터 분리된 Sesquiterpene Lactone의 세포독성 (Cytotoxity of Sesquiterpene Lactones from Leaves of Helianthus tuberosus L.)

  • 최현규;강연복;유시용;나민균;이승호
    • 생약학회지
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    • 제43권1호
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    • pp.6-9
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    • 2012
  • The $CH_2Cl_2$ soluble part of the leaves of Helianthus tuberosus L. (Compositae) exhibited a potent cytotoxic activity against the cultured human tumor cell lines including A-549, SK-OV-3, SK-MEL-2, XF498 and HCT-15 in vitro. Bioassaydirected fractionation of the $CH_2Cl_2$ soluble part of this plant led to the isolation of four cytotoxic sesquiterpene lactones having ${\alpha}$-methylene-${\gamma}$-lactone ring in the molecule. On the basis of physical and spectral evidences, their structures were characterized as ${\Delta}^{4,15}$-isoatripliciolide tiglate (1), ${\Delta}^{4,15}$-isoatripliciolide methacrylate (2), budlein A isobutylate (3) and budlein A tiglate (4). The ${\Delta}^{4,15}$-isoatripliciolide tiglate (1) showed the most potent cytotoxic activity ($0.26{\mu}M<ED_{50}<2.16{\mu}M$) against all of the cell lines tested.

수종의 암세포주와 치은섬유아세포에서 방사선의 양과 분할조사에 따른 세포활성도와 독성의 변화에 관한 연구 (AN EXPERIMENTAL STUDY ON THE CHANGE OF THE RADIOSENSITIVITY OF SEVERAL TUMOR CELL LINES AND PRIMARY CULTURED GINGIVAL FIBROBLAST)

  • 이삼선;유동수
    • 치과방사선
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    • 제27권1호
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    • pp.107-122
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    • 1997
  • Radiation sensitivity data was generated for two human cancer cell lines(KB, RPMI 2650) and human primary gingival fibroblast was tested three times using a viable cell number counting with a hemocytometer, MTT(3-[4,5-Dimethylthiazol2-yl]-2,5-diphenyl tetrazolium bromide) assay, and LDH(Lactate dehydrogenase) assay. Single irradiation of 2, 4, 6, 10, 15, 20Gy were applied to the tumor cell lines and the primary cultured gingival fibroblast The two fractions of 4Gy and 10Gy were seperated with a 4 hour time interval. The irradiation was done with 241.5cGy/min dose rate using /sup 137/Cs MK cell irradiator at room temperature. The obtained results were as followed : 1. There was significantly different viable cell numbers as the amount of radiation dose on the tested cells were cell number counted with a hemocytometer. In fractions, there were more viable cells remaining. 2. Phase-contrast microscopically, radiation-induced morphologic changes were pronounced on the tumor cells, however, almost no differences on the gingival fibroblast. 3. There was significantly different absorbance at 2Gy on RPMI 2600, 4Gy on KB and GF in MTT assay. In fractions, the absorbance was significantly higher on KB. 4. The level of extracellular LDH activity in the experimental group was significantly higher in the 2-4Gy than the control group. 5. The total level of extracellular and intracellular LDH activity was decreased as increased amounts of radiation dose was applied.

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Ginsenoside Rg1 suppresses cancer cell proliferation through perturbing mitotic progression

  • Hong, Jihee;Gwon, Dasom;Jang, Chang-Young
    • Journal of Ginseng Research
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    • 제46권3호
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    • pp.481-488
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    • 2022
  • Background: Although the tumor-suppressive effects of ginsenosides in cell cycle have been well established, their pharmacological properties in mitosis have not been clarified yet. The chromosomal instability resulting from dysregulated mitotic processes is usually increased in cancer. In this study, we aimed to investigate the anticancer effects of ginsenoside Rg1 on mitotic progression in cancer. Materials and methods: Cancer cells were treated with ginsenoside Rg1 and their morphology and intensity of different protein were analyzed using immunofluorescence microscopy. The level of proteins in chromosomes was compared through chromosomal fractionation and Western blot analyses. The location and intensity of proteins in the chromosome were confirmed through immunostaining of mitotic chromosome after spreading. The colony formation assays were conducted using various cancer cell lines. Results: Ginsenoside Rg1 reduced cancer cell proliferation in some cancers through inducing mitotic arrest. Mechanistically, it inhibits the phosphorylation of histone H3 Thr3 (H3T3ph) mediated by Haspin kinase and concomitant recruitment of chromosomal passenger complex (CPC) to the centromere. Depletion of Aurora B at the centromere led to abnormal centromere integrity and spindle dynamics, thereby causing mitotic defects, such as increase in the width of the metaphase plate and spindle instability, resulting in delayed mitotic progression and cancer cell proliferation. Conclusion: Ginsenoside Rg1 reduces the level of Aurora B at the centromere via perturbing Haspin kinase activity and concurrent H3T3ph. Therefore, ginsenoside Rg1 suppresses cancer cell proliferation through impeding mitotic processes, such as chromosome alignment and spindle dynamics, upon depletion of Aurora B from the centromere.

폐종양 환자에서 방사선치료에 의한 폐손상 (Radiation-induced Pulmonary Damage in Lung Cancer Patients)

  • 정수미;최일봉;강기문;김인아;신경섭
    • Radiation Oncology Journal
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    • 제11권2호
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    • pp.321-330
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    • 1993
  • Purpose: A retrospective analysis was performed to evaluate the incidence of radiation induced lung damage after the radiation therapy for the patients with carcinoma of the lung. Method and Materials: Sixty-six patients with lung cancer (squamous cell carcinoma 27, adenocarcinoma 14, large cell carcinoma 2, small cell carcinoma 13, unknown 10) were treated with definitive, postoperative or palliative radiation therapy with or without chemotherapy between July 1987 and December 1991. There were 50 males and 16 females with median age of 63 years (range: 33~80 years). Total lung doses ranged from 500 to 6,660 cGy (median 3960 cGy) given in 2 to 38 fractions (median 20) over a range or 2 to 150 days (median in days) using 6 MV or 15 MV linear accelerator. To represent different fractionation schedules of equivalent biological effect, the estimated single dose (ED) model, $ED=D{\dot}N^{-0.377}{\dot}T^{-0.058}$ was used in which D was the lung dose in cGy, N was the number of fractions, and T was the overall treatment time in days. The range of ED was 370 to 1357. The endpoint was a visible increase in lung density within the irradiated volume on chest X-ray as observed independently by three diagnostic radiologists. Patients were grouped according to ED, treatment duration, treatment modality and age, and the percent incidence of pulmonary damage for each group was determined. Result: In 40 of 66 patients, radiation induced change was seen on chest radiographs between 11 days and 314 days after initiation of radiation therapy. The incidence of radiation pneumonitis was increased according to increased ED, which was statistically significant (p=0.001). Roentgenographic changes consistent with radiation pneumonitis were seen in $100\%$ of patients receiving radiotherapy after lobectomy or pneumonectomy, which was not statistically significant. In 32 patients who also received chemotherapy, there was no difference in the incidence of radiation induced change between the group with radiation alone and the group with radiation and chemotherapy, among the sequence of chemotherapy No correlation was seen between incidence of radiation pneumonitis and age or sex. Conclusions: The occurrence of radiation pneumonitis varies. The incidence of radiation pneumonitis depends on radiation total dose, nature of fractionation, duration of therapy, and modifying factors such as lobectomy or pneumonectomy.

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생리활성분획 추적방법을 통한 오미자 추출물의 항염증 활성 분석 (Activity-guided Screening of Anti-inflammatory Compounds from the Hexane Extracts of Schisandra chinensis Fruit)

  • 최희정;최영환;백선용;김봉선;안순철;이문수;윤식
    • 생명과학회지
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    • 제23권2호
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    • pp.311-318
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    • 2013
  • 예로부터 한방이나 민간요법에서 다양한 약리효능을 가진 약제로 알려져 있는 오미자 열매로부터 항염증 활성을 갖는 유용한 물질을 동정하고자 본 연구를 수행하였다. 먼저 오미자의 hexane 추출물로부터 38개의 분획물을 분리한 다음 각 분획의 항염증 활성을 측정하였다. 그 결과 오미자 분획물 중 생리활성이 높은 SCKH1을 선택하여 활성분획 추적방법을 통해 SCKH1PAIBPB을 분리하였다. SCKH1PAIBPB는 VCAM-1, MCP-1, IL-6 및 IL-8의 발현을 감소시키며, 혈관내피세포와 단핵구 사이의 부착능을 억제시킨다는 사실을 확인할 수 있었다. 따라서 본 연구를 통해 규명된 오미자 분획물 및 SCKH1PAIBPB의 항염증효과 뿐만 아니라 혈관내피세포 증식 및 생존능 촉진작용을 응용하여, 다양한 허혈성질환 뿐만 아니라 염중성질환의 예방 및 치료에 활용할 수 있을 것으로 사료된다.

고량강 추출물의 암세포증식 저해 효과 (Anti-proliferative Effect of the Rhizome Extract of Alpinia officinarum on Cultured Human Tumor Cell Lines)

  • 이호성;차미란;최춘환;최상운;김영섭;김영균;김영호;연규환;유시용
    • 생약학회지
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    • 제39권4호
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    • pp.347-351
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    • 2008
  • The methanol (MeOH) extract of the rhizome of Alpinia officinarum Hance (Zingiberaceae) demonstrated a potent inhibition on the proliferation of cultured human tumor cell lines such as MES-SA (human uterine carcinoma cell line), MESSA/DX5 (multidrug resistant subline of MES-SA), HCT-15 (human colorectal adenocarcinoma cell line), HCT15/CL02 (multidrug resistant subline of HCT15). The MeOH extract was fractionated into four portions by serial solvent partition, ie., methylene chloride (CH2Cl2) soluble part, ethylacetate (EtOAc) soluble part, n-butanol (BuOH) soluble part and remaining water layer. Among them, the $CH_2Cl_2$ soluble part of the extract exhibited a most potent inhibition on the proliferation of tested tumor cell lines. Bioassay-guided fractionation of the $CH_2Cl_2$ soluble part led to the isolation of five diarylheptanoid and two flavonoid constituents, i. e., galangin (1), 7-(4"-hydroxy-3"-methoxyphenyl)-1-phenylhept-4-en-3-one (2), 1,7-diphenyl-5-hydroxy-3-heptanone (3), trans,trans-1-(3'-methoxy-4'-hydroxyphenyl)-7-phenyl-5-ol-4,6-dien-3-heptanone (4), 5-methoxy-7-(4"-hydroxy-3"-methoxyphenyl)-1-phenyl-3-heptanone (5), kaempferide (6), 5-hydroxy-7-(4"-hydroxy-3"-methoxyphenyl)-1-phenyl-3-heptanone (7). Structures of the isolated active components (1 - 7) were established by chemical and spectroscopic means.

Chemopreventive Potential of an Ethyl Acetate Fraction from Curcuma Longa is Associated with Upregulation of p57kip2 and Rad9 in the PC-3M Prostate Cancer Cell Line

  • Rao, K.V.K.;Samikkannu, T.;Dakshayani, K.B.;Zhang, X.;Sathaye, S.S.;Indap, M.A.;Nair, Madhavan P.N.
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권3호
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    • pp.1031-1038
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    • 2012
  • Background: Turmeric ($Curcuma$ $longa$) has been shown to possess anti-inflammatory, antioxidant and antitumor properties. However, despite the progress in research with $C.$ $longa$, there is still a big lacuna in the information on the active principles and their molecular targets. More particularly very little is known about the role of cell cycle genes $p57^{kip2}$ and Rad9 during chemoprevention by turmeric and its derivatives especially in prostate cancer cell lines. Methods: Accordingly, in this study, we have examined the antitumor effect of several extracts of $C.$ $longa$ rhizomes by successive fractionation in clonogenic assays using highly metastatic PC-3M prostate cancer cell line. Results: A mixture of isopropyl alcohol: acetone: water: chloroform: and methanol extract of $C.$ $longa$ showed significant bioactivity. Further partition of this extract showed that bioactivity resides in the dichloromethane soluble fraction. Column chromatography of this fraction showed presence of biological activity only in ethyl acetate eluted fraction. HPLC, UV-Vis and Mass spectra studies showed presence three curcuminoids in this fraction besides few unidentified components. Conclusions: From these observations it was concluded that the ethyl acetate fraction showed not only inhibition of colony forming ability of PC-3M cells but also up-regulated cell cycle genes $p57^{kip2}$ and Rad9 and further reduced the migration and invasive ability of prostate cancer cells.

면양(緬羊) Squamous Cell Carcinoma의 세포추출액(細胞抽出液)중에 함유된 종양특이(腫瘍特異) 및 면역기능저하물질(免疫機能低下物質) (Tumour Specific and Immunosuppressive Components in Soluble Cell Extracts from Ovine Squamous Cell Carcinoma)

  • 전무형
    • 농업과학연구
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    • 제13권2호
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    • pp.299-310
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    • 1986
  • 3M KCl을 이용하여 면양 squamous cell carcinoma 및 정상면양 조직(組織)에서 획득(獲得)한 추출액(抽出液)의 성상을 sephadex column chromatography, lymphocyte blastogenicity assay, acid dissociation method 및 gradient polyacrylamide gel electrophoresis 등을 이용하여 연구하였다. Sephadex column chromatography에서 얻은 5개의 종양분획(腫瘍分劃)중 분획(分劃)III은 가장 종양특이(腫瘍特異)한 항원(抗原)을 보유하고, 분획(分劃)V lymphocyte reactivity를 저하(低下)시키는 성분을 함유하고 있었다. 각 분획(分劃)을 분자량(分子量) > 100,000, 10,000~100,000 및 < 10,000의 3군(群)으로 분리한 바 종양특이항원(腫瘍特異抗原) 및 면역저지물질(免疫沮止物質)은 10,000~100,000분획(分劃)에서 나타났다. 분획(分劃)I tumour specific antigen - antibody complex를 내포하고 있음이 밝혀졌다. Gradient gel electrophoresis를 이용하여 분획(分劃)을 더욱 세분했을 때 분획(分劃)III은 Slice No 4~6에서 종양특이물질(腫瘍特異物質)이 인정되었고, 분획(分劃)V Slice No 9~11에서 면역기능저하물질(免疫機能低下物質)이 있음이 밝혀졌다.

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자궁경부 액상세포검사의 수기 검사법에 대한 고찰 - $SurePath^{TM}$ 검사법을 준용한 수기 검사법으로 - (Evaluation of the Manual Method of Liquid-Based Uterine Cervicovaginal Cytology - By The Manual Method Based on $SurePath^{TM}$ Methodology)

  • 박종명;장진욱;임소여;서인수;이종기
    • 대한세포병리학회지
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    • 제15권2호
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    • pp.86-91
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    • 2004
  • Liquid-Based Uterine Cervicovaginal Cytology is known to be a sensitive and effective screening method for cervical neoplasm $MonoPrep^{TM},\;ThinPrep^{TM},\;and\;SurePath^{TM}$ methods have been recently used as Liquid-Based Uterine Cervicovaginal Cytology techniques, and the $SurePath^{TM}$ method has been used in Sung-Yoon Reference Laboratory since 2003. The goal of Liquid-Based Uterine Cervicovaginal Cytology is to separate cervical epithelial cells from non-target cells, red blood cells and neutrophils. This report describes a study which evaluated cellularity, stainablilty, and cellular changes of epithelial cels in samples processed using a manual technique as compared to samples processed using $SurePath^{TM}$ automated method. The samples processed by means of a manual technique contained a cellularity of epithelial cells similar to that of the samples processed using the $SurePath^{TM}$ automated method. In addition, we compared variable density gradient reagents, including dextran, dextrose, and sucrose, to $SurePath^{TM}$ gradient media in order to evaluate cell fractionation and cellularity of epithelial cells. 10% dextran of gradient media shows good fractionation. The samples processed with 10% dextran demonstrated sufficient cellularity of epithelial cells and shows the fewest cellular changes. In conclusion, using a manual technique on these samples is easier to read than those results obtained using the $SurePath^{TM}$ automated method.

호알칼리성 Coryeform bacteria TU-19가 생산하는 세종류의 균체외 단백질분해호소의 정제 (Purification of Three Extracellular Proteases from Alkalophilic Coryneform Bacteria TU-19)

  • 최명철;양재섭;강선철
    • Applied Biological Chemistry
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    • 제38권6호
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    • pp.534-540
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    • 1995
  • 토양에서 분리된 호알칼리성 coryneform bacteria TU-19는 적어도 세종류의 단백질분해효소(Protease I, II, III)를 생성분비하였다. 이 균주의 효소생산과 관련된 배양조건을 조사해본 결과 최적온도 및 pH는 각각 $30^{\circ}C$와 10.0인 것으로 나타났다. 이틀 배양된 배양액으로부터 이 효소들을 정제하기 위해서 ammonium sulfate fractionation, gel filtration 및 QAE-Sephadex column chromatography 등을 순차적으로 행하였다. 그 결과 이들 세종류의 효소는 SDS-PAGE pattern으로 평가했을 때 single band로 순수 정제되었으며, 각각의 분자량은 120, 80, 45 kDa이었다. 정제된 효소의 특성을 살펴보면 Pretense I과 II의 최적 pH와 온도는 각각 $10.5,\;45^{\circ}C$이었으며, Protease III는 11.0과 $50^{\circ}C$로 나타났다. 또한 이들 세효소는 10 mM PMSF 농도에서 효소활성을 완전히 상실하였으며, pCMB, 1,10-phenanthroline, IAA, EDTA에는 별 영향이 없는 것으로 보아 serine protease의 일종으로 생각된다.

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