• 제목/요약/키워드: A-549 cell line

검색결과 203건 처리시간 0.029초

Glycosyltransformation of ginsenoside Rh2 into two novel ginsenosides using recombinant glycosyltransferase from Lactobacillus rhamnosus and its in vitro applications

  • Wang, Dan-Dan;Kim, Yeon-Ju;Baek, Nam In;Mathiyalagan, Ramya;Wang, Chao;Jin, Yan;Xu, Xing Yue;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • 제45권1호
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    • pp.48-57
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    • 2021
  • Background: Ginsenoside Rh2 is well known for many pharmacological activities, such as anticancer, antidiabetes, antiinflammatory, and antiobesity properties. Glycosyltransferases (GTs) are ubiquitous enzymes present in nature and are widely used for the synthesis of oligosaccharides, polysaccharides, glycoconjugates, and novel derivatives. We aimed to synthesize new ginsenosides from Rh2 using the recombinant GT enzyme and investigate its cytotoxicity with diverse cell lines. Methods: We have used a GT gene with 1,224-bp gene sequence cloned from Lactobacillus rhamnosus (LRGT) and then expressed in Escherichia coli BL21 (DE3). The recombinant GT protein was purified and demonstrated to transform Rh2 into two novel ginsenosides, and they were characterized by nuclear magnetic resonance (NMR) techniques and evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide assay. Results: Two novel ginsenosides with an additional glucopyranosyl (6→1) and two additional glucopyranosyl (6→1) linked with the C-3 position of the substrate Rh2 were synthesized, respectively. Cell viability assay in the lung cancer (A549) cell line showed that glucosyl ginsenoside Rh2 inhibited cell viability more potently than ginsenoside Rg3 and Rh2 at a concentration of 10 μM. Furthermore, glucosyl ginsenoside Rh2 did not exhibit any cytotoxic effect in murine macrophage cells (RAW264.7), mouse embryo fibroblasts cells (3T3-L1), and skin cells (B16BL6) at a concentration of 10 μM compared with ginsenoside Rh2 and Rg3. Conclusion: This is the first report on the synthesis of two novel ginsenosides, namely, glucosyl ginsenoside Rh2 and diglucosyl ginsenoside Rh2 from Rh2 by using recombinant GT isolated from L. rhamnosus. Moreover, diglucosyl ginsenoside Rh2 might be a new candidate for treatment of inflammation, obesity, and skin whiting, and especially for anticancer.

사철쑥 추출물의 면역세포의 생육증진 및 세포독성 (Enhanced Immune Activity and Cytotoxicity of Artemisia capillaris Thunb. Extracts against Human Cell Lines)

  • 이미경;최근표;류이하;이강윤;유창연;이현용
    • 한국약용작물학회지
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    • 제12권1호
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    • pp.36-42
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    • 2004
  • 사철쑥 추출물은 정상폐세포인 HEL299 세포주에 $0.5\;g/{\ell}$ 이하의 농도로 투여시는 정상세포 생존율을 80% 이상으로 유지시켜 정상세포에 대한 안정성이 유지되었으며, 암세포 생육억제활성은 증류수 및 에탄올 추출물 모두 $0.5\;mg/m{\ell}$ 이상의 농도에서 유방암세포주인 MCF7에 대하여 88%이상, 폐암세포주인 A549에 대하여서는 모든 추출물에서 $0.5\;mg/m{\ell}$의 농도 이상에서 65%이상의 저해를 나타내었다. 또한 간암세포주인 Hep3B에 대하여는, 에탄올 추출물이 87% 이상, 위암세포주인 AGS에 대하여 증류수 추출물이 약 85% 이상의 높은 암세포 생육억제활성을 나타내었다. 또한 정상세포독성에 대한 암세포의 억제율을 나타내는 selectivity에 있어서 각 추출물이 $0.1\;mg/m{\ell}$에서 $1.0\;mg/m{\ell}$의 농도에서 1.5 이상의 수치를 나타내었다. B와 T 세포주의 생육촉진 실험 결과 배양 기간이 증가함에 따라 세포의 생육이 촉진되었으며, 배양 5일째 $0.5\;mg/m{\ell}$의 농도에서 B세포의 생장을 최고 1.2배 이상 증가, T-cell을 $1.2{\sim}1.5$배 이상 생장을 증가시켰으며, 배양 6일째 에탄올 추출물이 IL-6를 $67\;pg/m{\ell},\;TNF-{\alpha}$를 물 추출물이 $68\;pg/m{\ell}$의 분비 하였다.

황기(黃芪)의 재배 년수에 따른 면역 및 항산화 활성 연구 (Studies on Immunomodulatory and Antioxidant Activities of Astragali membranacei Radix according to the Cultivated Years)

  • 정철
    • 대한한방피부미용학회지
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    • 제1권1호
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    • pp.53-90
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    • 2005
  • Purpose: Contents of astragaloside I, II and IV, cytotoxicity, anticancer activity, immunomodulatory activity and antioxidant capacity were to be compared as a function of the cultivated years as one, three, five and seven years. Method: Major components of Astragali membranacei Radix were separated as astragaloside I, astragaloside II, astragaloside IV by HPLC analysis. Cytotoxicity and anticancer activities were measured by MTT and SRB assay. For immunomodulatory activity, the secretion of IL -6 and $TNF-{\alpha}$, NK cell activation and macrophage activation were observed as well as kinetics of responding to human T cells by a microphysiometer. In vitro antioxidant activities were measured by several radical scavenging activities of superoxide anion radican, DPPH, LDL and linoleic acid. For in vivo activity, the activation of SOD, GSH-px, catalase, ALDH and ADH was measured as well the relative weight of liver. Result : 1. For HPLC analysis, the contents of all of astragaloside I, astragaloside II, astragaloside IV were in order of three, five, one and seven years. 2. The cytotoxicity of normal human lung cell line, HEL299 showed lower than 18% in adding 0.25 mg/ml, and 28.9% in adding 1.0 mg/ml of water extract of seven year root. For methanol extracts, three year root showed highest cytotoxicity as 35.2 % and there was no difference between the cultivated years. 3. For anticancer activities, methanol extracts of one and three year roots showed relatively high inhibition of human stomach cancer cells, AGS, breast cancer cells, MCF-7, lung cancer cells, A549 and liver cancer cell, Hep3B as well as high selectivities. 4. The water extract of seven year root could yield high secretion of IL-6 from both human Band T cells while the methanol extracts of three and five year roots secreted high amounts of IL-6 and $TNF-{\alpha}$ from both Band T cells. 5. As a result of in vitro antioxidant activities, both water and methanol extracts from five and seven year roots showed high activities for superoxide anion radical scavenging activity, inhibiting linoleic acid peroxide and contents of total phenols. 6. For in vivo tests, Mn-SOD and GSH-px activities and weight of liver were better in adding seven year root. For ALDH activity one year root was better and for ADH activity five year root. Overall speaking, seven year root showed relatively better antioxidant activities. Conclusion:There was difference of the contents of astragaloside I, astragaloside II, astragaloside IV according to cultivation year. Methanol extract showed better activities of anticancer and immune activation rather than water extract Interestingly enough, for methanol extracts, overall activities were improved as the cultivation year increased. There might be further investigation required for the clinical uses of the results as several biological activities varied according to the cultivated year of Astragali membranacei Radix.

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Comparison of Inhibitory Effect of 17-DMAG Nanoparticles and Free 17-DMAG in HSP90 Gene Expression in Lung Cancer

  • Mellatyar, Hassan;Akbarzadeh, Abolfazl;Rahmati, Mohammad;Ghalhar, Masoud Gandomkar;Etemadi, Ali;Nejati-Koshki, Kazem;Zarghami, Nosratallah;Barkhordari, Amin
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권20호
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    • pp.8693-8698
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    • 2014
  • Background: Up-regulation of hsp90 gene expression occurs in numerous cancers such as lung cancer. D,L-lactic-co-glycolic acid-poly ethylene glycol-17-dimethylaminoethylamino-17-demethoxy geldanamycin (PLGA-PEG-17DMAG) complexes and free 17-DMAG may inhibit the expression. The purpose of this study was to examine whether nanocapsulating 17DMAG improves the anti cancer effect over free 17DMAG in the A549 lung cancer cell line. Materials and Methods: Cells were grown in RPMI 1640 supplemented with 10% FBS. Capsulation of 17DMAG is conducted through double emulsion, then the amount of loaded drug was calculated. Other properties of this copolymer were characterized by Fourier transform infrared spectroscopy and H nuclear magnetic resonance spectroscopy. Assessment of drug cytotoxicity on the grown of lung cancer cell line was carried out through MTT assay. After treatment, RNA was extracted and cDNA was synthesized. In order to assess the amount of hsp90 gene expression, real-time PCR was performed. Results: In regard to the amount of the drug load, IC50 was significant decreased in nanocapsulated(NC) 17DMAG in comparison with free 17DMAG. This was confirmed through decrease of HSP90 gene expression by real-time PCR. Conclusions: The results demonstrated that PLGA-PEG-17DMAG complexes can be more effective than free 17DMAG in down-regulating of hsp90 expression by enhancing uptake by cells. Therefore, PLGA-PEG could be a superior carrier for this kind of hydrophobic agent.

물이끼 추출물의 식물화학적 성분 및 항암활성 연구 (Phytochemical Constituents and Anticancer Activity of Sphagnum palustre Extract)

  • 남정환;정진철;윤영호;홍수영;김수정;진용익;이예진;유동림;이경태;박희준
    • 한국자원식물학회지
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    • 제24권1호
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    • pp.40-47
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    • 2011
  • 본 연구에서는 척리 즉 물이끼를 이용한 고부가가치 기능성 소재를 개발하기 위하여 물이끼의 Ethanol 추출물을 이용한 항암활성과 식물화학적 성분연구를 수행 하였다. 3종의 암세포주(A549, HeLa, SK-OV-3)에 대하여 세포독성연구를 수행해 본 결과 폐암세포주인 A549에서 유의성 있는 결과를 보였다. 이 결과는 난치성 질환인 암을 치료하려는 목적 의약품 보다는 Chemopreventive agent로서의 예방 의학적 기능성 소재로 충분한 가치가 있음이 사료 되어지기에 식물화학적 성분연구를 실시하여 7종의 화합물(Comp.1 : Coumarin, Comp.2 : Caffeic acid, Comp.3 : Quercetin, Comp.4 : Astragalin, Comp.5 : Luteolin, Comp.6 : Chlorogenic acid, Comp.7 : Rutin) 를 분리하여 구조동정 하였다.

검정찰옥수수 종실 분쇄 정도에 따른 항산화 및 Cytotoxicity 활성 효과 (Effect of Particle Size on Antioxidant Activity and Cytotoxicity in Purple Corn Seed Powder)

  • 김정태;손범영;이진석;백성범;우관식;정건호;김미정;정광호;권영업
    • 한국작물학회지
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    • 제57권4호
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    • pp.353-358
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    • 2012
  • 본 연구에서는 검정찰옥수수 종실을 일반분쇄와 저온미세분쇄를 하여 입자크기에 따른 항산화 활성과 cytotoxicity를 평가하여 이용성 증진을 하고자 연구를 수행하였다. 본 연구의 결과를 요약하면 다음과 같다. 1. 분쇄 정도에 따른 평균값은 일반분쇄가 $473.7{\mu}m$, 저온미세분쇄 $17.2{\mu}m$이었으며 중간값은 $336.9{\mu}m$$13.4{\mu}m$를 나타내었다. 2. TEAC 활성 측정 결과 일반분쇄는 $3.87{\mu}mol$ TE/g로서 저온미세분쇄 $5.15{\mu}mol$ TE/g보다 낮은 활성을 보였다. FRAP 활성 측정에서는 저온미세분쇄가 $10.08{\mu}mol$ Fe(II)/g로 일반분쇄 $8.86{\mu}mol$ Fe(II)/g보다 높은 활성을 보였다 3. 간암 세포주(Hep-G2) 성장억제에 미치는 영향은 1 mg/ml 농도에서 검정찰옥수수 종실을 일반분쇄(31.48%)한 것보다 저온미세분쇄(27.41%)를 한 경우가 암세포 생장 억제 효과를 큰 것으로 나타났다. 4. 대장암 세포주인 HCT-116에서는 1 mg/ml에서 분쇄정도에 따라 큰 차이를 보이지 않았으며, 유방암 세포주(MCF-7)에서는 일반분쇄가 미세분쇄보다 높은 생장억제를 보였다.

발계(Smilax china L.) 열수 및 에탄올 추출물의 항산화 및 항암활성 (Antioxidant and anticancer properties of hot water and ethanol extracts from the roots of Smilax china L.)

  • 김예진;손대열
    • 한국식품저장유통학회지
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    • 제20권5호
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    • pp.691-698
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    • 2013
  • 식품 연구에서 흔히 사용되는 물과 주정(70% 에탄올)을 사용하여 추출된 발계(SCR)의 열수 추출물(SCRW)과 에탄올 추출물(SCRW)의 총 플라보노이드 및 폴리페놀 함량, 항산화 활성, 항암활성을 측정하였다. SCRW와 SCRE는 DPPH(SCRW; 72.9%, SCRE; 42.4%)와 ABTS(SCRW; 81.6%, SCRE; 50.3%) 자유 라디칼을 효과적으로 소거하였으며, 양성 대조군인 BHA(53.8% DPPH, 49.5% ABTS)보다 더 높은 소거활성을 나타냈다. 또한 인체 유래 신장 정상세포(HEK293)에 대해서는 높은 생존율을 나타내 독성이 없음을 확인한 반면, 인체 유래의 암세포(유방암세포; MCF-7, 폐암세포; A549, 위암세포; AGS)에 대해서는 대조군으로 사용된 항암제 CPA와 유사하거나 더 높은 세포성장 억제효과가 확인되었다. 특히 SCRE는 AGS에 대하여 $1,000{\mu}g/mL$ 농도에서 53.6%로 탁월한 위암 세포 성장저해 효과를 나타냈다. 본 연구 결과를 통해, 발계 추출물의 높은 항산화 및 항암 활성이 확인되어 약리활성에 대한 기초연구 자료로 제시 할 수 있을 것으로 사료되며, 기능성 소재로써의 이용 가치를 높일 수 있을 것으로 판단된다.

살구 추출물의 항산화성, 항돌연변이성 및 세포독성 효과 (Antioxidative, Antimutagenic and Cytotoxic Effects of Prunus armeniaca Extracts)

  • 유수정;김수현;전미선;오현택;최현진;함승시
    • 한국식품저장유통학회지
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    • 제14권2호
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    • pp.220-225
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    • 2007
  • 살구 에탄올 추출물의 항산화 활성은 RC50값이 살구씨와 살구과육 에탄을 추출물의 경우 각각 $48.3{\mu}g$$43.9{\mu}g$으로서 강한 항산화 활성을 나타내었다. 살구 에탄을 추출물의 항돌연변이 효과의 검토는 Salmonella typhimurium의 변이주인 TA98과 TA100을 이용한 Ames test로 확인하였다. 그 결과 살구씨 및 과육 에탄올 추출물 자체의 돌연변이원성은 없었고 직접변이원인 MNNG에 대해 시료농도 $200{\mu}g/plate$에서 살구씨와 살구과육 에탄올 추출물 각각에서 TA100이 69.4% 및 65.9%의 억제효과를 나타내었다. 같은 농도에서 4NQO에 대해서는 살구과육 에탄을 추출물의 경우TA98과TA100이 각각45.9% 및 44.3%의 억제효과를 나타내었다. 암세포 성장억제효과를 검토한 결과 살구씨 에탄올 추출물 4mg/mL첨가 시 A549, AGS, MCF-7, HeLa 및 Hep3B에서 각각 63.7, 56, 86.3, 78 및 53.7%의 억제 효과를 보였다. 살구과육 에탄을 추출물 4mg/mL 첨가시 위암세포 AGS에서 58.0% 억제효과를 보인 반면 모든 암세포에서 72.8%이상의 높은 억제효과를 나타내었다. 이러한 암세포에 대한 높은 억제 효과에 비해 인간정상신장세포 293에 대해서는 37.2% 이하의 생육 억제율을 나타냄으로서 정상세포에 대해서는 낮은 독성효과를 가짐을 알 수 있었다.

Simultaneous Blockage of Epidermal Growth Factor Receptor and Cyclooxygenase-2 in a Human Xenotransplanted Lung Cancer Model

  • Mu, Xiao-Yan;Dong, Xue-Li;Sun, Jie;Ni, Yu-Hua;Dong, Zhang;Li, Xi-Li;Sun, Er-Lian;Yi, Zhou;Li, Gao
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권1호
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    • pp.69-73
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    • 2014
  • The effects of erlotinib combined with celecoxib in a lung cancer xenograft model were here explored with a focus on possible mechanisms. A xenotransplanted lung cancer model was established in nude mice using the human lung cancer cell A549 cell line and animals demonstrating tumour growth were randomly divided into four groups: control, erlotinib, celecoxib and combined (erotinib and celecoxib). The tumor major axis and short diameter were measured twice a week and after 40 days tissues were collected for immunohistochemical analyses of Bcl-2 and Bax positive cells and Western-blotting analyses for the epidermal growth factor recepto (EGFR), P-EGFR, and cyclooxygenase-2 (COX-2). Tumor size in the combined group was smaller than in the others (p<0.01) and the percentage of Bcl-2 positive cells was fewer in most cases (p<0.01), while that of Bax positive cells was greater than in the erlotinib and celecoxib groups (P>0.05). Western blotting showed decreased expression of P-EGFR and COX-2 with both erlotinib and celecoxib treatments, but most pronouncedly in the combined group (P<0.05). Simultaneous blockage of the EGFR and COX-2 signal pathways exerted stronger growth effects in our human xenotransplanted lung cancer model than inhibition of either pathway alone. The anti-tumor effects were accompanied by synergetic inhibition of tumor cell apoptosis, activation of p-EGFR and expression of COX-2.

삼백초 추출물의 항암활성과 카드뮴에 대한 독성억제효과 (Effects of Saururus Chinensis Baill Extracts on Anticancer Activity and Cadmium Induced Cytotoxicity)

  • 박승정;유현주;최희석;서병윤;양선호;김영훈;정재열;이기남
    • 대한예방한의학회지
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    • 제8권2호
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    • pp.81-98
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    • 2004
  • 1. The cell viability was determined by MTT method. Their cytotoxic activities against three cancer cell lines such as A549, MDA-MB-231 and SNU-C4 cell line were tested. Among them, The methanol extract of Saururus chinensis Baill. showed the strongest cytotoxic effect against SNU-C4 cells. These results suggest that the methanol extract of Saururus Chinensis Baill. possessed a potential antitumorous agent. 2. In vitro the antitoxic activity of ethanol extract of Saururus Chinensis Bail on NIH 3T3 fibroblasts was evaluate by the MTT (3-(4,5-dimethyl- thiazol-2-yl)-2,5-diphenyl-2H-tetra캐lium bromide) and SRB (sulforhodamine B protein) assays. The number of NIH 3T3 fibroblasts were increased and tend to regenerate. These results suggest that Saururus Chinensis Bail extract retains a potential antitoxic activity. 3. Complexation of Cd (II) ion with ligands such as quercetin has been determined by UV-vis spectrophotometric method in tris buffer solution at various pH. It was found that only 1 : 1 Cd-complex is formed by the interaction between catechol moiety in ring B and cadmium [Cd(II)] in aqueous solution. The spectral parameters for Cd-complex were determined by Beer's law at various pH. It has shown that 1 : 1 Cd-complex has a maximum absorbance and red shift by the alkaline pH.

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