• Title/Summary/Keyword: Cell assay

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Anti-oxidant, Anti-inflammatory and Anti-cancer Effect of Methanol Extract of Pogostemon cablin (광곽향 메탄올 추출물의 항산화, 항염증 및 암세포 증식 억제 효과)

  • Yun, Seung Geun;Jin, Soojung;Jeong, Hyun Young;Yun, Hee Jung;Do, Mi young;Kim, Byung Woo;Kwon, Hyun Ju
    • Journal of Life Science
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    • v.25 no.1
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    • pp.44-52
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    • 2015
  • In the present study, the substance that show anti-proliferation of cancer cells as well as anti-oxidant and anti-inflammatory effect was searched. As a results, the methanol extract of Pogostemon cablin (P. cablin), is a well-known herb for traditional medicine in Korea and China for treating the digestive disorders, less of appetite, vomiting and diarrhea, inhibited the growth of various cancer cells such as A549, HepG2, MCF7 and HT29 cells. Cytotoxic effect of methanol extraction of P. cablin was excellent in A549 cells. P. cablin extract induced cell cycle arrest at G1 phase of A549 in a dose dependent manner. And it induced phosphorylation of p38 and Cdc25A and reduced expression of Cdc25A, Cdks, Cyclins and phospho-Retinoblastoma (Rb) proteins. Therefore, P. cablin extract seems to act through the p38 - Cdc25A - Cdk - Cyclin - Rb pathway in A549 cells. In addition, P. cablin extract showed anti-oxidant effect by DPPH free radical scavenging assay and anti-inflammation effect by inhibition of lipopolysaccharide (LPS)-induced nitric oxide (NO) in RAW 264.7 cells in a dose-dependent manner. Taken together, these results suggest that P. cablin may be used as not only candidate materials for anti-cancer, anti-inflammatory and anti-oxidant, moreover, it would be possible utilized in various health functional food materials.

Studies on Analysis Method of T-2 Toxin by ELISA (ELISA에 의한 T-2 toxin의 분석법에 관한 연구)

  • 오유진;장성재;윤여표
    • Journal of Food Hygiene and Safety
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    • v.3 no.2
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    • pp.65-73
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    • 1988
  • T-2 toxin is one of mycotoxins produced by fungi such as Fusarium spp. and possesses a potent cytotoxicity to eukaryotic cell. The contamination of mycotoxins in cereals and feedstuffs is one of the great concerns in health authorities. Therefore, the development of the specific, sensitive and simplified analysis method for T -2 toxin is required. During more than ten years, several chemical and biological analysis methods were proposed and applied for the detection and quantification of T-2 toxin. TLC, GLC-FID and GC-MS are widely employed, but these methods required numerous clean-up procedures before analysis, and the detection limit for T-2 toxin is more than 10 ppb. Biological analysis methods with dermal tissues and cultured cells are not specific to T-2 toxin, since T-2 toxin and other related derivatives possess a similar toxicological activity although their relative activity is different each otber. Based on tbe specific reaction between antibody and antigen, the authors tried to introduce the immunochemical methods for determination of T-2 toxin. The enzyme-linked immunosorbent assay method using monoclonal antibody for T-2 toxin was applied to analyse T-2 toxin. The detection limit of T-2 toxin by ELISA method was 0.1 ppb. The correlation between ELISA and GC-MS method on these samples was very high. ELISA method developed for the detection and quantification of T -2 toxin in this paper possesses simplicity, high sensitivity and specific for T-2 toxin. Furthermore, the ELISA method with T-2 toxin monoclonal antibody was an excellent tool for the screening of Fusarium spp. which was suspected to produce T-2 toxin.

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Effects of Fermented Milk Containing Herb Extract from Acanthopanax divaricatus var. albeofructus and Codonopsis Ianceolata on the Immune Status of Mouse (흰털오가피와 더덕 추출물을 첨가한 발효유 급여가 마우스의 면역기능에 미치는 영향)

  • Lim, Sang-Dong;Seong, Ki-Seung;Kim, Kee-Sung;Han, Dong-Un
    • Food Science of Animal Resources
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    • v.27 no.1
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    • pp.95-101
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    • 2007
  • We have investigated the immunomodulatory activity of water extracts of Acanthopanax divaricatus var. alveofructus in male ICR mice. Mice were administrated with fermented milk containing freeze-dried extract 3 mg/Kg (A), 9 mg/kg (B), 27 mg/Kg (C) per body weight with A. divaricatus var. alveofructus (loots : leaves : stem) : Codonopsis lanceolata = (5 : 2 : 1.5) : 1.5 for 7 and 10 weeks, respectively. Body weight, relative organ weight, cellularity of lymphoid organs, plaque- forming cell (PFC) assay, agglutination (AGG) test and lymphoproliferation were examined in various groups of animals. Any significant differences of body weight gain were not recorded in the tested ICR mice. There was significant different (p<0.05) in the spleen index in B group of 10 weeks and C group of 7 weeks fed mouse. The thymus gain weight was increased during administration of the extract, but there was no significant increase on other organs gain. Humoral immunity as measured by PFC showed more decreased PFC level in 10 weeks than in 7 weeks. In the HT, A. divaricatus var. albeofructus extract also showed a significant increase (p<0.05) in C group of 10 weeks. Administration of extracts from A. divaricatus var. albeofructus increased significantly in the production of IgG antibodies on the mice immunized with SRBC in B group of 7 and 10 weeks (p<0.05).

Anti-osteoarthritis effects of Pomegranate, Eucommiae cortex and Achyranthis radix extracts on the primary cultured rat articular chondrocytes

  • Choi, Beom-Rak;Ku, Sae-Kwang;Kang, Su-Jin;Park, Hye-Rim;Sung, Mi-Sun;Lee, Young-Joon;Park, Ki-Moon
    • Journal of Society of Preventive Korean Medicine
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    • v.21 no.3
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    • pp.87-98
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    • 2017
  • Objectives : The objective of present study is to evaluate anti-arthritic effects of dried pomegranate concentrate powders (PCP), Eucommiae Cortex aqueous (EC) and ethanolic (ECe) extracts, Achyranthis Radix aqueous (AR) and ethanolic (ARe) extracts on the primary cultured rat articular chondrocytes. Methods : MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium Bromide) assay was performed cytotoxic effect of test substances. In addition, anti-inflammatory effects were also observed on the lipopolysaccaride (LPS) treated chondrocytes through prostaglandin $E_2\;(PGE_2)$ production and 5-lipoxygenase (LPO) activities, and inhibitory effects on metalloproteinase (MMP)-2 and MMP-9 activities were observed on the recombinant human interleukin $(rhIL)-1{\alpha}$ treated chondrocytes with their extracellular matrix (ECM) related mRNA expressions - collagen type II, SOX9 and aggrecan. Results : As results, ECe and ARe showed obvious cytotoxicity against primary cultured rat articular chondrocytes at a dose level of 10 mg/ml, respectively. However, no obvious cytotoxic effects of PCP, EC and AR were demonstrated at a dose level of 10 mg/ml, on the primary cultured rat articular chondrocytes. In addition, treatment of LPS $50{\mu}g/ml$ induced significant increases of $PGE_2$ contents and 5-LPO activities indicating inflammatory responses of the primary cultured rat articular chondrocytes, and also decreases of cell viabilities, increases of MMP-2 and MMP-9 activities with decreases of extracellular matrix (ECM) related collagen type II, SOX9 and aggrecan mRNA expressions were observed by treatment of $rhIL-1{\alpha}$ 50 ng/ml, suggesting damages on the primary cultured rat articular chondrocytes and related ECM degradations. However, these inflammatory responses and related ECM degradations were inhibited by pretreatment of all test substances, in order of PCP > ECe > ARe > EC > AR, and $rhIL-1{\alpha}$ induced chondrocytes deaths are inhibited by treatment in order of PCP > EC > AR > ECe > ARe. Conclusions : Taken together, it is expected that mixed formulation of PCP as main components with appropriate proportion of EC and AR as additional components will be achieved a potent alternative medicinal food for osteoarthritis.

Studies on antimicrobial agent produced by lactobacillus acidophilus (Lactobacillus acidophilus가 생산한 항균물질(抗菌物質)에 관한 연구(硏究))

  • Kim, Dong-shin
    • Korean Journal of Veterinary Research
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    • v.24 no.2
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    • pp.149-162
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    • 1984
  • The research was conducted(1) to confirm the agent(s) responsible for the antimicrobial activity contained in the fermented tomato juice with L. acidophilus(2) to extract and purify the antimicrobial agent(s)(3) to find the biological, physical and chemical properties of the agent(s). The following results were obtained and summarized as followings; 1. The agent responsible for the inhibitory activity was confirmed by both well assay method using fermented tomato juice with L. acidophilus and turbidimetric technique using the cell-free filtrate or neutralized filtrate of tomato acidohilus culture and found exerted antimicrobial agent other than lactic acid. 2. The procedures of purification : The isolation and purification of antimicrobial agent from the lyophilized acidophilus tomato culture were carried out by (1) methanol extraction (2) acetone extraction, (3) Sephadex G-50 gel filtration (4) paper chromatography and (5) thin layer chromatography. 3. The biological, physical and chemical properties of antimicrobial agent: The biological, physical, chemical properties of the purified antimicrobial agent were: (1) The antimicrobial activity was strong against test organisms; Bacillus subtilis(ATCC 6633), Escheichia coli(ATCC 25922), Staphylococcus aureus(ATCC 167), Pseudomonas fluorescens(KFCC 32394), Proteus vulgaris and Shigella dysenteriae. (2) The pH value of the agent was 2.0 and the inhibitory activity was lost when it was neutralized at 7.0 of pH and the agent was heat stable at $121^{\circ}C$ for 60 minutes. (3) The ultraviolet light absorption spectra of methanol-acetone extract and TLC fraction exhibited a maximum absorption at 260nm and 224nm respectively. (4) The most purified agent from TLC plate increased about 130-fold in activity. (5) The agent isolated from TLC plate was free from $H_2O_2$ or lactic acid. 4. Bioautographic assy: By means of bioautography of the agent on silica gel of TLC plate a strong inhibitory activity against B. subtilis was demonstrated. 5. Mass spectrometry: The agent obtained from TLC plate was analyzed by mass spectrometry which show the parent peak at m/e 264 suggesting the molecular weight of the compound and molecular group such as [$C_2H{^+}_4$], [CO], [CH=NH], [$C_3{H^}4_7$], [$\begin{array}{rcl}O\\{\parallel}\\CH_3-C\\\end{array}$], [$C_6-H{^+}_{11}$], [$C_5H{^+}_{11}$], [$\begin{array}{rcl}O\\{\parallel}\\C_5H_7-C^+\\\end{array}$] were suggested.

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Studies on the proliferative activity and the expression of c-myc and ras of preneoplastic hepatocytes in rats (랫드 전암 간세포의 증식성과 c-myc, ras의 발현에 관한 연구)

  • Cho, Ho-seong;Park, Nam-yong
    • Korean Journal of Veterinary Research
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    • v.40 no.1
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    • pp.117-129
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    • 2000
  • This study was carried out to examine proliferative activity and expression of c-myc oncoprotein and p2lras in normal and preneoplastic rat livers induced by an in vivo mid-term chemical carcinogenesis assay. Sixty, six-week-old male specific pathogen free Sprague-Dawley male rats were randomly divided into five groups. Group I was received a single intraperitoneal(IP) dose(200mg/kg) of diethylnitrosamine(DEN). Group 2(10 rats) was operated partial hepatectomy(PH) and Group 3 was received IP(200mg/kg) DEN, fed two weeks later with 500ppm of phenobarbital(PB). Group 4 was received IP(200mg/kg) DEN, fed two weeks later 500ppm(PB) and PH at week 3 after the onset of experiment. While group 5(20 rats) was not treated and used as a control group. All the rats were sacrificed at age 14 weeks except 10 rats from group 5 were sacrificed at the onset of experiment. Livers of all rats were examined for 5-bromo-2'-deoxyuridine(BrdU) incoporation, proliferating cell nuclear antigen(PCNA), silver-binding nucleolar organizer regions(AgNORs) counts per nucleus and expression of c-myc oncoprotein and p21ras. Both the number and area of the preneoplastic lesions were significantly(p<0.01) compared to other groups. A significant(p<0.01) increase in immunoreactive cells were detected in preneoplastic hepatocytes in Groups 3 and 4 by PCNA and BrdU immunohistochemical stain. The number of the positive cells were significantly(p<0.05) lower in normal 14-week-old rats than those of 6-week-old rats. The results showed that proliferative activity of the hepatocytes was increased by treatment with DEN, PH and PB. Meanwhile, AgNORs counts per nucleus were significantly(p<0.05) increased in the preneoplastic hepatocytes of rats in both groups 3 and 4. The expression of c-myc oncoprotein and p21ras were more readily localized within the hepatic preneoplastic lesions such as hyperplastic nodules. Especially, group 4 showed significantly (p<0.05) overexpressed levels compared to groups 1 and 3. These findings suggest that PCNA, BrdU and AgNORs are significantly increased and c-myc oncoprotein and p21ras are significantly overexpressed in hepatic preneoplastic lesions induced by mid-term carcinogenesis. So these parameters can be an effective markers for hepatic prencoplastic lesions.

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Isolation of porcine reproductive and respiratory syndrome virus(PRRSV) in Korea (돼지생식기 및 호흡기증후군(Porcine Reproductive and Respiratory Syndrome ; PRRSV) 바이러스의 국내분리주 작성에 관한 연구)

  • Kweon, Chang-hae;Kwon, Byung-joon;Lee, Han-jung;Cho, Jae-jin;Hwang, Eui-kyung;Shin, Jin-ho;Yoon, Yong-dhuk;Kang, Yung-bai;An, Soo-hwan;Kim, Yong-hee;Huh, Won;Jun, Moo-hyung;Wensvoort, G.
    • Korean Journal of Veterinary Research
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    • v.34 no.1
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    • pp.77-83
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    • 1994
  • Three viral strains, causing CPE in porcine alveolar macrophage cell, were isolated from aborted fetus, serum from young pig showing blue-ear sign and lung of suspected pig, respectively. The differential diagnostic results showed no characteristics of Aujeszky's disease virus(ADV), hog cholera virus (HCV), Japanese encephalitis virus(JEV), porcine parvovirus(PPV) and encephalomyocarditis virus (EMCV). However, positive reactions were demonstrated by IFA using monospecific porcine antibodies against Lelystad virus. When the paired sera of experimentally inoculated swine with one of isolate, KPRRSV-l were tested by IPMA, the result indicated that the isolate was related to United States isolate than European LV.

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Genetic Toxicity Test of o-Nitrotoluene by Ames, Micronucleus, Comet Assays and Microarray Analysis

  • Lee, Eun-Mi;Lee, So-Youn;Lee, Woo-Sun;Kang, Jin-Seok;Han, Eui-Sik;Go, Seo-Youn;Sheen, Yhun-Yong;Kim, Seung-Hee;Park, Sue-Nie
    • Molecular & Cellular Toxicology
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    • v.3 no.2
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    • pp.107-112
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    • 2007
  • o-Nitrotoluene is used to synthesize artificial dyes and raw materials of urethane resin. In this study, we have carried out in vitro genetic toxicity tests and microarray analysis to understand the underlying mechanisms and the mode of action of toxicity of onitrotoluene. TA1535 and TA98 cells were treated with o-nitrotoluene to test its toxicity by basic genetic toxicity test. Ames and two new in vitro micronucleus and COMET assays were applied using CHO cells and L5178Y cells, respectively. In addition, microarray analysis of differentially expressed genes in L5178Y cells in response to o-nitrotoluene was analyzed using Affymatrix genechip. The result of Ames test was that o-nitrotoluene treatment did not increase the mutations both in base substitution strain TA1535 and in frame shift TA98. o-Nitrotoluene has not increased micronuclei in CHO cells. But onitrotoluene increased DNA damage in L5178Y cell. Two-hundred two genes were initially selected as differentially expressed genes in response to o-nitrotoluene by microarray analysis and forty four genes among them were over 2 times of log fold changed. These forty four genes could be candidate biomarkers of genetic toxic action of o-nitrotoluene related to induction of mutation and/or induction of micronuclei and DNA damage. Further confirmation of these candidate markers related to the DNA damage will be useful to understand the detailed mechanism of action of o-nitrotoluene.

Evaluation of Sleep Comfort for Indoor Thermal Environment based on the Physiological Signal Analysis (생리신호 분석을 이용한 실내온열환경에서의 수면 쾌적성 평가)

  • 이낙범;임재중;금종수;이구형;최호선
    • Science of Emotion and Sensibility
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    • v.3 no.2
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    • pp.75-84
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    • 2000
  • 문명의 발달과 함께 수면부족으로 인한 여러 가지 스트레스와 질환이 증가하게 되어 최근 수면연구에 대한 관심이 증가하고 있다. 본 연구는 다양한 온열환경 조건에서의 쾌적한 수면을 위한 온열환경을 제시하기 위해, 5명의 여성 피험자를 대상으로 22$^{\circ}C$, 26$^{\circ}C$, 3$0^{\circ}C$의 일정온도 조건과 $25^{\circ}C$에서 1시간 후와 2시간 후에 각각 1, 2$^{\circ}C$를 상승시켜주는 변동온도 조건하에서 수면생리신호를 측정하였다. 그리고, 수면단계 평가를 이용하여 총 수면시간, 깊은 수면의 비율, 그리고 최초 수면시작 시간에서 최초의 서파 수면이 나타나기까지의 지연시간 등의 수면효율을 평가하였다. 그 결과, 일정온도 조건에서는 26$^{\circ}C$에서 총 수면시간(466.7$\pm$10.25분)과 깊은 수면의 비율(33.1$\pm$4.95%)은 타 조건에 비해 높게 나타났고, 최초 서파수면까지의 지연시간(9.8$\pm$3.33분)은 타 조건에 비해 낮게 나타나 쾌적한 수면을 위한 가장 적절한 온열조건임을 관찰할 수 있었다. 그리고 변동온도 조건에서는 4가지 온열조건간에는 큰 차이가 나타나지 않았지만, 모든 조건에서 일정온도 조건보다는 좋은 결과를 나타내었다. 또한 수면 중 신체 움직임과 설문 분석에서도 동일한 결과를 보였다. 본 연구를 통해, 수면생리신호를 이용한 수면 쾌적성 분석은 수면의 질적인 상태를 관찰하는데 매우 적합한 파라메터를 도출할 수 있으며, 여러 가지 수면환경 조건을 평가하는데 매우 유용한 지표가 될 수 있음을 보였다.e results suggest that hCG treatment at 7 days after insemination could be used to increase the pregnancy rate of embryo transfer, and transfer, and only the recipients with PUN concentration of <12 mg/dl were influenced by treatment with hCG./TEX>이었으며, 이는 화성 기원을 지시한다.sucrose를 이용한 2단계 희석이 수정란의 생존성을 향상시키는 것으로 나타났다. 또한 발육 단계별 생존성에 있어서는 발육이 진전된 확장배 반포 시에 동결하는 것이 배반포기에 동결하는 것 보다 유리한 것으로 나타났다.ody를 사용하여 flow cytometery해석을 실시하는 한편 125I-hGH binding assay에 의하여 hGH binding activity를 측정하였다. 최종적으로 GH signal transduction의 target genedf으로 알려져 있는 serine protease inhibitor 2.1(Spi 2.1) gene의 promotor activity를 검토한 결과 hGHR을 transfect한 CHO Cell에 있어서 hGH의 농도에 의존적으로 증가되었다. 따라서 본 실험에서 cloning한 cDNA hGHR는 native hGHR와 같은 기능을 가지는 것으로 판명되었다.것으로 판명되었다..ments of that period left both in Japan and Korea. "Hyojedo" in Korea is supposed to have been influenced by the letter design. Asite- is also considered to

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Beta-4 Integrin Transfection, Cloning and Functional Assay in Squamous Cell Carcinoma (Beta-4 Integrin 유전자 주입, 클로닝과 편평상피암에서의 Beta-4 Integrin 기능에 관한 연구)

  • Kim Young-Min;Carey Tomas E.
    • Korean Journal of Head & Neck Oncology
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    • v.13 no.2
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    • pp.169-179
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    • 1997
  • 서론 : Laminin의 수용기로 알려진 Integrin $\alpha6\beta4$의 세포내 표현 정도는 편평상피암을 위시한 여러 악성종양의 전이능력 및 예후와 밀접한 상관관계가 없다고 알려져 있다. 이 Integrin은 Laminin과 같은 세포와 리간드와 결합하면 상피세포의 기저막 지주 구조물인 hemidesmosome의 세포체질 요소(cytoskeletal element)와 연관되어 그 결과 세포의 기저막과 세포내 케라틴을 연결하는 역할을 한다. Integrin $\alpha6\beta4$는 구조적으로 다른 많은 integrin들과 달리 $\beta$4의 세포질내 영역(cytoplasmic domain)이 특징적으로 크다. 이 세포질내 영역 $\beta$4 integrin의 기능은 아직 밝혀지지 않고 있으나 아마 세포 성장의 신호전달 및 악성종양의 특징인 침윤 전이에 관련할 것으로 보아지고 있다. 재료 및 방법: 저자들은 우선 $\beta$4 integrin의 wild type s-DNA와 $\beta$4 세포질내 영역(cytoplasmic domain) 및 $\beta$4의 tyrosine 인산화 반응 부위가 각각 결손된 c-DNA를 PCR을 통하여 합성하여 pRc/CMV 벡터에 삽입한 후 원래 $\beta$4 integrin의 발현이 결집된 인간 방광암 세포에 Calcium phosphate precipitation 방법으로 주입(transfection)시켜 형질변환된 세포를 면역형광법, Flow cytometry 및 Immunoprecipitation 방법으로 클로닝하여 wild type $\beta$4-full length(Clone FL), truncated $\beta$4-cytoplasmic domain(C1one CD), 및 mutated $\beta$4-tyrosine phosphorylation site (Clone M)을 얻었다. 암 세포의 부착 및 침투 능력의 기능적 연구로 모노 클로날 항체와 fibronectin, laminin, Matrigel을 단백질 기질로 사용하였으며 결과 비교를 위하여 pRc/CMV 벡터만 주입시켰던 클로운과 방광암 세포주를 $\beta$4 integrin 음성 대조군으로 또한 이 Integrin의 높은 발현을 보이는 두경부 편평상피암 세포주를 양성 대조군으로 이용하였다. 결과 : 세포부착능력에 있어서 온전한 $\beta$4 cytoplasmic domain이 존재하는 클로운이 laminin에 강한 부착능력을 보였으나 fibronectin의 부착정도는 $\beta$4 integrin의 표현정도와 관계없이 모든 클로운에서 비슷하였다. Matrigel을 투과하는 암세포 침윤 능력에서는 $\beta$4 integrin의 표현이 존재하는 클로운들이 투과 능력이 높았으나 세포외 리간드가 없는 control membrane을 사용하였을 때와 비교하여 투과능력의 차이를 보이지 않았다. 결론 : 유전자 주입(transfection) 방법으로 integrin의 다양한 클로운의 합성이 가능하여 이 Integrin의 암 세포의 부착 및 침투 능력에서의 기능을 규명 할 수 있게 한다. $\beta$4 integrin은 편평상피 암세포의 부착에 있어서 세포외 리간드 laminin과 특이 결합하여 부착 능력을 높이는 중요한 역할을 하며 편평상피 암세포의 침투에 있어서는 $\beta$4 integrin의 표현이 침투 능력을 높이는 역할을 하나 이때에는 laminin과 같은 리간드와의 특이 결합에 의존하지는 않는 것으로 사료된다.

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