• 제목/요약/키워드: Population Growth Inhibition

검색결과 93건 처리시간 0.031초

인간 대장암 HT-29 세포에서 제주조릿대의 세포사멸 효과 (Apoptotic Effect of Sasa quelpaertensis Nakai in Human Colon Cancer HT-29 Cells)

  • 변지희;김민영
    • 생명과학회지
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    • 제24권9호
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    • pp.1012-1018
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    • 2014
  • 제주조릿대(Sasa quelpaertensis Nakai)는 한라산에 넓게 분포되어 자생하는 식물로 최근 연구에서 항염증, 항당뇨, 항산화, 항암 효능을 가지는 것으로 알려져 있으나 대장암에서의 항암 효능 및 그에 따른 mechanism에 대해서는 명확히 밝혀지지 않았다. 본 연구에서는 인간 대장암 HT-29 세포를 대상으로 제주조릿대에 의한 항암작용과 기전에 대해 조사하였다. 제주조릿대에 의한 HT-29 세포의 증식 억제가 apoptosis 유도와 연관성이 있음을 DNA fragmentation와 flow cytometry 분석에 의한 sub-G1기의 세포빈도의 증가로 확인하였다. 제주조릿대에 의한 apoptosis 유발은 HT-29 세포의 S arrest 현상을 동반하였을 뿐만 아니라 발생한 산화질소의 증가와 anti-apoptotic factor인 IAP family (survivin, XIAP, cIAP-1, cIAP-2) 발현이 감소함으로써 촉진되었음을 확인할 수 있었다. 이러한 결과들은 제주조릿대가 대장암에 대한 치료제로서의 사용 가능성을 확인할 수 있었지만 이를 입증하기 위해서는 더 자세한 항암기전에 관한 연구가 진행되어야 한다고 사료된다.

Systemic Approaches Identify a Garlic-Derived Chemical, Z-ajoene, as a Glioblastoma Multiforme Cancer Stem Cell-Specific Targeting Agent

  • Jung, Yuchae;Park, Heejoo;Zhao, Hui-Yuan;Jeon, Raok;Ryu, Jae-Ha;Kim, Woo-Young
    • Molecules and Cells
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    • 제37권7호
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    • pp.547-553
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    • 2014
  • Glioblastoma multiforme (GBM) is one of the most common brain malignancies and has a very poor prognosis. Recent evidence suggests that the presence of cancer stem cells (CSC) in GBM and the rare CSC subpopulation that is resistant to chemotherapy may be responsible for the treatment failure and unfavorable prognosis of GBM. A garlic-derived compound, Z-ajoene, has shown a range of biological activities, including anti-proliferative effects on several cancers. Here, we demonstrated for the first time that Z-ajoene specifically inhibits the growth of the GBM CSC population. CSC sphere-forming inhibition was achieved at a concentration that did not exhibit a cytotoxic effect in regular cell culture conditions. The specificity of this inhibitory effect on the CSC population was confirmed by detecting CSC cell surface marker CD133 expression and biochemical marker ALDH activity. In addition, stem cell-related mRNA profiling and real-time PCR revealed the differential expression of CSC-specific genes, including Notch, Wnt, and Hedgehog, upon treatment with Z-ajoene. A proteomic approach, i.e., reverse-phase protein array (RPPA) and Western blot analysis, showed decreased SMAD4, p-AKT, 14.3.3 and FOXO3A expression. The protein interaction map (http://string-db.org/) of the identified molecules suggested that the AKT, ERK/p38 and $TGF{\beta}$ signaling pathways are key mediators of Z-ajoene's action, which affects the transcriptional network that includes FOXO3A. These biological and bioinformatic analyses collectively demonstrate that Z-ajoene is a potential candidate for the treatment of GBM by specifically targeting GBM CSCs. We also show how this systemic approach strengthens the identification of new therapeutic agents that target CSCs.

Runx3 inhibits endothelial progenitor cell differentiation and function via suppression of HIF-1α activity

  • SO-YUN CHOO;SOO-HYUN YOON;DONG-JIN LEE;SUN HEE LEE;KANG LI;IN HYE KOO;WOOIN LEE;SUK-CHUL BAE;YOU MIE LEE
    • International Journal of Oncology
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    • 제54권4호
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    • pp.1327-1336
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    • 2019
  • Endothelial progenitor cells (EPCs) are bone marrow (BM)-derived progenitor cells that can differentiate into mature endothelial cells, contributing to vasculogenesis in the blood vessel formation process. Runt-related transcription factor 3 (RUNX3) belongs to the Runt domain family and is required for the differentiation of specific immune cells and neurons. The tumor suppressive role of RUNX3, via the induction of apoptosis and cell cycle arrest in a variety of cancers, and its deletion or frequent silencing by epigenetic mechanisms have been studied extensively; however, its role in the differentiation of EPCs is yet to be investigated. Therefore, in the present study, adult BM-derived hematopoietic stem cells (HSCs) were isolated from Runx3 heterozygous (Rx3+/-) or wild-type (WT) mice. The differentiation of EPCs from the BM-derived HSCs of Rx3+/- mice was found to be significantly increased compared with those of the WT mice, as determined by the number of small or large colony-forming units. The migration and tube formation abilities of Rx3+/- EPCs were also observed to be significantly increased compared with those of WT EPCs. Furthermore, the number of circulating EPCs, defined as CD34+/vascular endothelial growth factor receptor 2 (VEGFR2)+ cells, was also significantly increased in Rx3+/- mice. Hypoxia-inducible factor (HIF)-1α was upregulated in Rx3+/- EPCs compared with WT EPCs, even under normoxic conditions. Furthermore, in a hindlimb ischemic mouse models, the recovery of blood flow was observed to be highly stimulated in Rx3+/- mice compared with WT mice. Also, in a Lewis lung carcinoma cell allograft model, the tumor size in Rx3+/- mice was significantly larger than that in WT mice, and the EPC cell population (CD34+/VEGFR2+ cells) recruited to the tumor was greater in the Rx3+/- mice compared with the WT mice. In conclusion, the present study revealed that Runx3 inhibits vasculogenesis via the inhibition of EPC differentiation and functions via the suppression of HIF-1α activity.

대청호 하류에서 환경요인과 Anabaena cylindrica 생장억제세균의 계절별 분포 변화 (Seasonal Variations of Environmental Factors and Distribution of Anabaena cylindrica Growth-Inhibiting Bacteria in the Lower Daechung Reservoir)

  • 이정호;김철호
    • 생태와환경
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    • 제33권2호통권90호
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    • pp.128-135
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    • 2000
  • 수화의 소멸과정에서 수계에 분포하는 세균의 역할을 확인하기 위하여 대청호의 주요 정점에서 환경요인과 종속영양세균 군집, Anabaena cylindrica 생장억제세균의 수층별 연중 변화를 조사하였다. 평균수심은 정점 1에서 25.5m, 정점 2에서 15.0m, 정점 3에서는 12.3m였다. 모든 정점에서 수층별 온도는 하절기를 중심으로 하는 온대지방 수계에서의 전형적인 변화를 나타내었다. 용존산소량은 하절기에 불규칙하게 변화하긴 했지만 수온과 비교적 반비례적인 양상으로 변화하였다. 대체적으로 모든 정점에서 5월 초순까지 점차 감소하던 용존산소량은 이후에는 급격한 변화를 나타내다가 9월 중순 이후에는 점차 증가하는 양상을 보였으며, 특히 이러한 변화양상은 표층에서 더욱 뚜렷하였다. 용존산소량이 불규칙적으로 큰 변화를 보이는 하절기에는 다른 수층에 비하여 표층에서 용존산소량이 매우 높았다. 표층에서의 클로로필 a농도는 하절기에 변동폭이 크게 나타났으며, 용존산소량이 높은 시기에 비교적 높은 수치를 나타내었다. 종속영양세균의 분포도는 모든 수층에서 봄부터 가을까지는 대체로 높게 유지되다가 수온이 급격히 하락하는 9월 이후에는 매우 낮은 개체군 크기를 보였다. 특히 모든 표층에서 이러한 변화는 두르러졌다. Anabaena cylindrica 생장억제세균은 6월 이전에는 거의 분포하지 않다가 7월에 분포도가 큰 폭으로 증가하였고, 이후부터는 정점별로 상이한 분포 변화를 보였다. 모든 정점에서 A. cylindrica생장억제세균의 분포값이 높았던 7월과 10월에는 표층보다는 중층과 하층에서 더 많은 생장억제세균이 검출되었는데, 이러한 결과는 숙주인 A. cylindrica의 활력이 상대적으로 떨어질 때 억제세균의 활성이 높아지는 것을 암시한다. 이상의 결과로부터 자연수계에서 세균은 남조세균 분포에 대한 조절자로서의 역할을 하며, 특히 남조세균의 활력이 떨어질 때 그 역할이 더욱 두드러진다는 사실을 알 수 있었다.

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Shalf Life Enhancement of Minimally Processed Fruits and Vegetables

  • Kim, Dong-Man
    • 한국식품저장유통학회:학술대회논문집
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    • 한국식품저장유통학회 1993년도 정기총회 및 제3차 학술발표회
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    • pp.6-9
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    • 1993
  • According to changes in population, economic conditions, life-stile and eating habits, the frui ts and vegetables market wi 11 be shi fted from processed (i. e. , canned) to fresh. Undressed fresh produce, consisting of washed, disinfected and peeled fruits and vegetables that either sliced or grated, are currently increased in demand by retail and institutional market which use them as salad components or in ready-to use foods, Main attributes of minimally processed fruits and vegetables are convenience and fresh-like quality. Minimally processed Products readily deteriorate in quality, especially color and texture, as a result of endogeneous enzyme enhanced respiration and microorganisms which lead to reduced shelf Iife. According to changes in population, economic conditions, life-stile and eating habits, the frui ts and vegetables market wi 11 be shi fted from processed (i. e. , canned) to fresh. Undressed fresh produce, consisting of washed, disinfected and peeled fruits and vegetables that either sliced or grated, are currently increased in demand by retail and institutional market which use them as salad components or in ready-to use foods, Main attributes of minimally processed fruits and vegetables are convenience and fresh-like quality. Minimally processed Products readily deteriorate in quality, especially color and texture, as a result of endogeneous enzyme enhanced respiration and microorganisms which lead to reduced shelf Iife. Thus. to prevent these undesirable changes , val'ious techniques such as controlled atmosphere (CA) storage, modified atmosphere OIA) storage, including vacuum packaging have been receiving considerable attention, Although milch research has been done to find optimal conditions for whole intact frui ts and vegetables, only limi ted information is avai lable on fresh cut. and other minimally processed products. 81 iced frui ts exhibi t increas~d ethylene production and respiration compal'ed to whole f, 'uits during distribution in response to tissue damage. As a result, accelerated senescence and enzymatic browning OCCUI', Recent l'esearch on minimally processed fl'uits and vegetables has mainly focused on methods to inhibit browning, due to ban on use of sulfur dioxide, In order to retard or prevent these physiological changes, val'ious al ternatives, reducing agents. acidulants, chelating agents and inol'ganic sal ts have been evaluated for use on fresh cut fl'ui ts. Al though some agents were effective replacement for sulfur dioxide. consum$\textregistered$I'S demandless use of chemical on such products. Shel~ life of minimally processed products has been extended by inhibition of metabolic reactions associated with loss of quality and by inhibition of aerobic spoilage caused by wide variety of microorganisms. Appl ication of ~I.-\ packaging, including vacuum packaging, retards the rate of respiration, prevents growth of aerobic spoilage organisms, inhibits oxidation and color deterioration. Tissue softening is another major problem in minimally processed products because enzymes re 1 a ted to ce 11 wa 11 degrada t i on are not inactivated. Various treatments have been investigated for retardation of the softening of sliced products. Some studies have concentrated on the application of an active packaging system with ~I, l. packaging and calcium infi 1 tration as possible measures to retain firmness of processed products. In my opinion, one important step for production of minimally processed frui ts wi th favorabl e color of cut surface and firm texture is the selection of better cultivar. As the view, changing tendency of fresh color by apple cultivars and relationship between the tendency and PPO activity will be discussed in the seminar. In addition to the topic, research result on quality enhancement of fresh apple slices by heat shock treatment will be introduced.

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저온저장중 Oregano(Origanum vulgare L.)가 Eschetichia coli O157:H7과 Staphylococcus aureus 196E의 생존에 미치는 영향 (Effect of Oregano (Origanum vulgare L.) on the Survival of Escherichia coli ) O157:H7 and Staphylococcus aureus 196E during Cold Storage)

  • 박찬성;박금순;김미림
    • 한국식품조리과학회지
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    • 제13권4호
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    • pp.440-447
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    • 1997
  • 향신료인 oregano(Origanum vulgare L.)를 액체배지에 첨가하여 2종류의 식중독세균(Escherichia coli O157:H7과 Staphylococcus aureus 196E)의 증식과 저온저장중 생존에 미치는 항균효과를 조사하였다. 저농도(0~2%, w/v)의 oregano를 액체배지(TSB)에 첨가하여 E. coli와 S. aureus를 $10^{6}$~$10^{7}$ cells/$m\ell$가 되게 접종한 후 35$^{\circ}C$에서의 증식과 냉장(5$^{\circ}C$)및 냉동(-2$0^{\circ}C$) 저장중 생존억제효과를 생균수의 변화로서 조사하였다. 35$^{\circ}C$에서의 E. coli의 증식은 1% 이내의 향신료 농도에서는 증식이 저해되지 않았으나, 2%의 oregano존재하에서는 짧은 유도기를 거친 후에 증식이 시작되었다. 정상기 이후의 사멸기에서는 첨가한 oregano의 농도가 높을수록 빠른 사멸속도를 나타내었다. 35$^{\circ}C$에서의 S. aureus의 증식은 0.1%의 경우에는 증식이 저해되지 않았으며 0.3%와 0.5%의 oregano 존재하에서는 긴 유도기를 거친 후에 증식이 시작되었으나 1~2%의 경우에는 계속적으로 생균수가 감소되었다. 5$^{\circ}C$에서 냉장한 경우에는 0.5%이상의 oregano에 의해 E. coli와 S. aureus 모두 우수한 항균효과를 나타내어 생존이 크게 억제되고 저장말기에 사멸하였다. -2$0^{\circ}C$에 동결저장하였을 때, E. coli는 저장기간이 길어질수록, 첨가한 향신료의 농도가 높을수록 생존억제효과가 증대되었으나 S. aureus는 저장초기에 급격히 감소한 후 거의 일정수준을 유지하였으며 첨가한 향신료의 농도에 따른 생균수의 차이는 나타나지 않았다. 동결저장에서 0.1%의 oregano를 첨가했을 때 E. coli는 대조구의 생균수에 비해 1/3~l/7로 생균수가 감소하였으나 S. aureu는 대조구에 비해 1/18~l/46로 감소하여 S. aureus에서 더 우수한 항균효과를 나타내었다.

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Field Studios of In-situ Aerobic Cometabolism of Chlorinated Aliphatic Hydrocarbons

  • Semprini, Lewts
    • 한국지하수토양환경학회:학술대회논문집
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    • 한국지하수토양환경학회 2004년도 총회 및 춘계학술발표회
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    • pp.3-4
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    • 2004
  • Results will be presented from two field studies that evaluated the in-situ treatment of chlorinated aliphatic hydrocarbons (CAHs) using aerobic cometabolism. In the first study, a cometabolic air sparging (CAS) demonstration was conducted at McClellan Air Force Base (AFB), California, to treat chlorinated aliphatic hydrocarbons (CAHs) in groundwater using propane as the cometabolic substrate. A propane-biostimulated zone was sparged with a propane/air mixture and a control zone was sparged with air alone. Propane-utilizers were effectively stimulated in the saturated zone with repeated intermediate sparging of propane and air. Propane delivery, however, was not uniform, with propane mainly observed in down-gradient observation wells. Trichloroethene (TCE), cis-1, 2-dichloroethene (c-DCE), and dissolved oxygen (DO) concentration levels decreased in proportion with propane usage, with c-DCE decreasing more rapidly than TCE. The more rapid removal of c-DCE indicated biotransformation and not just physical removal by stripping. Propane utilization rates and rates of CAH removal slowed after three to four months of repeated propane additions, which coincided with tile depletion of nitrogen (as nitrate). Ammonia was then added to the propane/air mixture as a nitrogen source. After a six-month period between propane additions, rapid propane-utilization was observed. Nitrate was present due to groundwater flow into the treatment zone and/or by the oxidation of tile previously injected ammonia. In the propane-stimulated zone, c-DCE concentrations decreased below tile detection limit (1 $\mu$g/L), and TCE concentrations ranged from less than 5 $\mu$g/L to 30 $\mu$g/L, representing removals of 90 to 97%. In the air sparged control zone, TCE was removed at only two monitoring locations nearest the sparge-well, to concentrations of 15 $\mu$g/L and 60 $\mu$g/L. The responses indicate that stripping as well as biological treatment were responsible for the removal of contaminants in the biostimulated zone, with biostimulation enhancing removals to lower contaminant levels. As part of that study bacterial population shifts that occurred in the groundwater during CAS and air sparging control were evaluated by length heterogeneity polymerase chain reaction (LH-PCR) fragment analysis. The results showed that an organism(5) that had a fragment size of 385 base pairs (385 bp) was positively correlated with propane removal rates. The 385 bp fragment consisted of up to 83% of the total fragments in the analysis when propane removal rates peaked. A 16S rRNA clone library made from the bacteria sampled in propane sparged groundwater included clones of a TM7 division bacterium that had a 385bp LH-PCR fragment; no other bacterial species with this fragment size were detected. Both propane removal rates and the 385bp LH-PCR fragment decreased as nitrate levels in the groundwater decreased. In the second study the potential for bioaugmentation of a butane culture was evaluated in a series of field tests conducted at the Moffett Field Air Station in California. A butane-utilizing mixed culture that was effective in transforming 1, 1-dichloroethene (1, 1-DCE), 1, 1, 1-trichloroethane (1, 1, 1-TCA), and 1, 1-dichloroethane (1, 1-DCA) was added to the saturated zone at the test site. This mixture of contaminants was evaluated since they are often present as together as the result of 1, 1, 1-TCA contamination and the abiotic and biotic transformation of 1, 1, 1-TCA to 1, 1-DCE and 1, 1-DCA. Model simulations were performed prior to the initiation of the field study. The simulations were performed with a transport code that included processes for in-situ cometabolism, including microbial growth and decay, substrate and oxygen utilization, and the cometabolism of dual contaminants (1, 1-DCE and 1, 1, 1-TCA). Based on the results of detailed kinetic studies with the culture, cometabolic transformation kinetics were incorporated that butane mixed-inhibition on 1, 1-DCE and 1, 1, 1-TCA transformation, and competitive inhibition of 1, 1-DCE and 1, 1, 1-TCA on butane utilization. A transformation capacity term was also included in the model formation that results in cell loss due to contaminant transformation. Parameters for the model simulations were determined independently in kinetic studies with the butane-utilizing culture and through batch microcosm tests with groundwater and aquifer solids from the field test zone with the butane-utilizing culture added. In microcosm tests, the model simulated well the repetitive utilization of butane and cometabolism of 1.1, 1-TCA and 1, 1-DCE, as well as the transformation of 1, 1-DCE as it was repeatedly transformed at increased aqueous concentrations. Model simulations were then performed under the transport conditions of the field test to explore the effects of the bioaugmentation dose and the response of the system to tile biostimulation with alternating pulses of dissolved butane and oxygen in the presence of 1, 1-DCE (50 $\mu$g/L) and 1, 1, 1-TCA (250 $\mu$g/L). A uniform aquifer bioaugmentation dose of 0.5 mg/L of cells resulted in complete utilization of the butane 2-meters downgradient of the injection well within 200-hrs of bioaugmentation and butane addition. 1, 1-DCE was much more rapidly transformed than 1, 1, 1-TCA, and efficient 1, 1, 1-TCA removal occurred only after 1, 1-DCE and butane were decreased in concentration. The simulations demonstrated the strong inhibition of both 1, 1-DCE and butane on 1, 1, 1-TCA transformation, and the more rapid 1, 1-DCE transformation kinetics. Results of tile field demonstration indicated that bioaugmentation was successfully implemented; however it was difficult to maintain effective treatment for long periods of time (50 days or more). The demonstration showed that the bioaugmented experimental leg effectively transformed 1, 1-DCE and 1, 1-DCA, and was somewhat effective in transforming 1, 1, 1-TCA. The indigenous experimental leg treated in the same way as the bioaugmented leg was much less effective in treating the contaminant mixture. The best operating performance was achieved in the bioaugmented leg with about over 90%, 80%, 60 % removal for 1, 1-DCE, 1, 1-DCA, and 1, 1, 1-TCA, respectively. Molecular methods were used to track and enumerate the bioaugmented culture in the test zone. Real Time PCR analysis was used to on enumerate the bioaugmented culture. The results show higher numbers of the bioaugmented microorganisms were present in the treatment zone groundwater when the contaminants were being effective transformed. A decrease in these numbers was associated with a reduction in treatment performance. The results of the field tests indicated that although bioaugmentation can be successfully implemented, competition for the growth substrate (butane) by the indigenous microorganisms likely lead to the decrease in long-term performance.

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부저병 원인균 Paenibacillus larvae 특이 유전자 분석을 통한 진단마커 발굴 (Identification of Diagnostic PCR Markers for Honeybee Foulbrood Disease from Specific Genes of Paenibacillus larvae)

  • 나한흠;김근철
    • 생명과학회지
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    • 제27권1호
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    • pp.67-71
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    • 2017
  • 부저병이란 Paenibacillus larvae감염에 의하여 꿀벌 유충의 괴사를 유도하는 질병이다. 우리나라에서는 2008년 봄, 국내에 처음으로 대량 발병 사례가 보고되었으며, 지속적인 2차 피해로 큰 후유증을 앓고 있다. 본 연구에서는 부저병을 효율적으로 관리할수 있는 진단방법을 조사하고자 하였다. 따라서 부저병의 원인균인 P. larvae에서 특이적으로 발현되고 있는 유전자들을 동정하고자 하였으며, 이 유전자들은 주로 부저병균의 독성을 유발하는 것으로 알려진 Toxin1, Toxin2A & 2B, SplA, CBP49, SevA&SevB 들이다. 이들은 1차 PCR 에서는 검출하기 어려웠지만, 2차 nested PCR방법을 이용하여 검출이 용이함을 알 수 있었다. 한편 여러가지 식물 추출물을 혼합한 배지에서 부저병균을 배양하였을 때, 부저병균의 성장저해와 일치하게 우리가 검증한 유전자들의 발현이 감소하는 것을 알 수 있었다. 이러한 결과들은 부저병 원인균의 특이 유전자들은 향후 PCR진단마커로서 활용 가능성이 있을 것으로 사료된다.

상황보리 추출물에 의한 p21 및 Bax 발현 증가와 caspase 활성화를 통한 U937 인체백혈병 세포의 apoptosis 유발 (Apoptotic Cell Death by Methanol Extract of Phellinus linteus-Barley Corn in Human Leukemic U937 Cells through Induction of p21 and Bax, and Activation of Caspase-3)

  • 박철;김현중;정경태;윤태경;최병태;이용태;박동일;최영현
    • 동의생리병리학회지
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    • 제21권5호
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    • pp.1226-1232
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    • 2007
  • Phellinus linteus is a well-known Oriental medicinal fungus that has various biological activities, including immunomodulatory and anti-tumor activities, the mechanisms of which are poorly understood. In the present study, we investigated the anti-proliferative activity of the methanol extract of P. linteus-Barley corn (MEPLB) in human lekemic U937 cells. It was found that exposure of U937 cells to MEPLB resulted morphological change and growth inhibition in a dose-dependent manner as measured by trypan blue count and MTT assay. Upon treatment with MEPLB, U937 cells developed many of the hallmark features of apoptosis, including condensation of chromatin and an increase in the sub-G1 population suggesting that the anti-proliferative effect of MEPLB is associated with the induction of apoptosis. The anti-proliferative and apoptotic effects of MEPLB were connected with a marked induction of the pro-apoptotic Bax and cyclin-dependent kinase (Cdk) inhibitor p21 in a p53-independent manner. Additionally, MEPLB treatment significantly induced the caspase-3 activity in U937 cells. Taken together, the present results suggest that apoptotic signals evoked by MEPLB in human leukemic U937 cells may converge caspase-3 activation through an up-regulation of Bax rather than a down-regulation of Bcl-2 or Bel-xL.

Cispatin 내성인 사람 위암 세포주 SNU-1의 복합약제내성 및 Verapamil의 효과 (Multidrug Resistance and Cytotoxicity of Anticancer Drug by Verapamil in Cisplatin Resistant Human Stomach Cancer Cell)

  • 손성권;김정희
    • Journal of Yeungnam Medical Science
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    • 제9권1호
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    • pp.75-89
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    • 1992
  • 복합약제내성암세포주의 출현은 암의 화학 요법에 있어서 중요한 문제점중의 하나이며 이 복합내성 암세포의 출현에 대한 정확한 기전은 아직 밝혀져 있지 않다. 본 실험에서는 약제내성 암세포의 다른 항암제에 대한 복합내성 형성정도와 calcium 길항제인 verapamil에 의한 내성극복 정도를 비교하고 내성 암세포의 세포막 단백질의 변화를 관찰하였다. 사람의 위암 세포주 SNU-1을 cisplatin 농도 $0.001{\mu}M$에서 시작하여 $10{\mu}M$까지 증가시켜 내성세포 SNU-1/$Cis_5$를 얻었으며, MTT assay로 세포성장을 관찰한 결과 doubling time은 SNU-1이 약 29 시간, SNU-1/$Cis_5$는 약 38시간으로 내성세포가 감수성세포보다 성장속도가 느린 것을 관찰하였다. 약제 감수성 검사를 위해서 4일간의 MTT assay로 대조군에 대한 50% 세포 생존시의 약제농도 $IC_{50}$를 비교하여 상대적 내성도 (Relative Resistance, RR)를 측정하였는데 cisplatin, 81.4; vinblastine, > 43.0; epirubicin, 22.9 ; dactinomycin, 16.0 ; etoposide, 15.0 ; vincristine, 9.2 ; adriamycin, 5.7 ; aclarubicin, 5.3으로 관찰되었고 그외의 약제인 cyclophosphamide, 5-fluorouracil, methotrexate, daunorubicin에서는 낮게 나타났다. $10{\mu}M$의 verapamil에 의한 내성 억제 효과는 vincristine, 13.1 ; epirubicin, 10.0 ; etoposide, 6.3 ; vinblastine, 4.4 ; dactinomycin, 3.6 ; daunorubicin, 2.4로 나타났다. Radioiodination을 이용한 SDS-PAGE로 SNU-1/$Cis_5$에서 항암제 내성과 관련된 것으로 여겨지는 51,400와 81,300 dalton의 막단백질의 발현을 관찰하였다.

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