• Title/Summary/Keyword: 3LL cells

Search Result 71, Processing Time 0.033 seconds

Effect of Multidrug Resistance Gene-1 (mdr1) Overexpression on In-Vitro Uptake of $^{99m}Tc$-sestaMIBl in Murine L1210 Leukemia Cells (백혈병 세포에서 Multidrug Resistance Gene-1 (mdr1)의 과발현이 $^{99m}Tc$-sestaMIBl 섭취에 미치는 영향)

  • Chun, Kyung-Ah;Lee, Jae-Tae;Lee, Sang-Woo;Kang, Do-Young;Sohn, Sang-Kyun;Lee, Jong-Kee;Chung, June-Key;Jun, Soo-Han;Lee, Kyu-Bo
    • The Korean Journal of Nuclear Medicine
    • /
    • v.33 no.2
    • /
    • pp.152-162
    • /
    • 1999
  • Purpose: To determine whether $^{99m}Tc$-MIBI is recognized by the multidrug resistant P-glycoprotein (Pgp), we have measured quantitatively $^{99m}Tc$-MIBI uptake in cancer cells. The effects of various Pgp reversing agents on cellular $^{99m}Tc$-MIBI uptake were also investigated in the presence of multidrug resistance gene-1 (mdr1 gene) overexpression. Materials and Methods: We measured percentage uptake of $^{99m}Tc$-MIBI at different incubation temperatures both in mdr1 positive and negative cells. The effects of verapamil, cyclosporin, and dipyridamole on cellular uptake of $^{99m}Tc$-MIBI were also evaluated with or without overex-pression of mdr1 gene in cultured murine leukemia Ll210 cells. Results: The mdr1 gene expressing cell lines were effectively induced in in vitro with continuous application of low-dose adriamycin or vincristine. Cellular uptake of $^{99m}Tc$-MIBI was higher in mdr1 negative Ll210 cells than those of mdr1 positive cells, and higher when incubated in $37^{\circ}C$ than $4^{\circ}C$. In the presence of verapamil, cyclosporin or dipyridamole, $^{99m}Tc$-MIBI uptake was increased upto 604% in mdr1 positive cells. Conclusion: Cellular uptake of $^{99m}Tc$-MIBI is lower in leukemia cells over-expressing mdr1 gene, and MBR-reversing agents increase cellular uptake. These results suggest that $^{99m}Tc$-MIBI can be used for characterizing Pgp expression and developing MDR-reversing agents in vitro.

  • PDF

Transmission electron microscopic ultrastructures of the male germinal cells of Fibricola seoulensis (Fibricola seoulensis 웅성생식세포의 투과전자현미경적 미세구조)

  • 손원목;이순형
    • Parasites, Hosts and Diseases
    • /
    • v.31 no.3
    • /
    • pp.183-192
    • /
    • 1993
  • A transmission electron microscopic study was performed to observe the ultrastructures of the male germinal cells and spermatozoa of Fibricola seoulensis. Spermatogonia were found in the periphery of the testis and characterized by large nuclei and comparatively little cytoplasms. Spermatocytes contained an oval to spherical nucleus. Their nuclear volume was little larger in comparative to that of cytoplasm, and the chromatin was comparatively little. The early spermatids were characterized by a great amount of cytoplasm, and numerous mitochondria encircled the nucleus. In a more advanced spermatids the electron-dense strands of chromatin appeared in the nucleus, and a pair of rootlet of the axoneme and a microtubule-organizing center (MTOC) were observed near the nucleus. The sectioned spermatozoa were found in the testis and the seminal vesicle. Their cross sectional views were divided into 6 types when they were distinguished on the basis of the morphology and components. The spermatozoa of F. seoulensis showed two flagella of 9+1 type axoneme.

  • PDF

Soluble isocitrate dehydrogenase plays a key role in obesity and hyperlipidemia

  • Koh, Ho-Jin;Lee, Su-Min;Huh, Tae-Lin
    • Proceedings of the Korean Society of Applied Pharmacology
    • /
    • 2003.11a
    • /
    • pp.5-7
    • /
    • 2003
  • NADPH is an essential co-factor for fat and cholesterol biosynthesis. However, the role of cytosolic NADP$\^$+/-dependent isocitrate dehydrogenase (IDPc), a putative NADPH producer, in the control of the fat and cholesterol metabolism has not been assessed. Here we report that increased or decreased IDPc expression in 3T3-Ll fat cells promoted or retarded adipogenesis, respectively. Furthermore, overexpression of IDPc in transgenic mice exhibited fatty liver, hypertriglyceridemia, hypercholesterolemia and obesity by increasing NADPH production leading to subsequent stimulation of acetyl-coenzyme A and malonyl-coenzyme A consumption. In contrast, administrations of a synthetic IDPc inhibitor, DAl1004, to ob/ob mice effectively reduced body weight with lowering cholesterol and triglyceride levels. In addition, a positive relationship (${\gamma}$ = 0.69, $\rho$<0.0l) between plasma IDPc activity and body mass indexes was observed in 98 randomly-selected human volunteers. Our findings strongly indicate that NADPH produced by IDPc plays an important role in controlling body fat and lipid biosynthesis.

  • PDF

Gene Expression of Enzymes Related to Glutathione Metabolism in Anticancer Drug-resistant L1210 Sublines (항암제 내성 L1210세포의 Glutathione 대사 관련효소 유전자의 발현 양상)

  • Kim, Seong-Yong;Kim, Jae-Ryong;Kim, Jung-Hye
    • Journal of Yeungnam Medical Science
    • /
    • v.12 no.1
    • /
    • pp.32-47
    • /
    • 1995
  • Glutathione(GSH) has a very important role in detoxification of cells and is closely related to antitumor drug-resistance of cancer cells. In order to evaluate the importance of glutathione metabolism in the drug-resistant cancer cells, the concentration of celluar GSH and activities of ${\gamma}$-glutamylcysteine synthetase(GCS), ${\gamma}$-glutamyl transpeptidase (GGT) and glutathione S-transferases(GST) in the adriamycin, vincristine, or cisplatin resistant L1210 (L1210AdR, L1210VcR, or L1210Cis) sublines were measured. Expression and amplification of GCS, GGT, and GST-${\pi}$ genes were also observed in the parent Ll210 and the drug-resistant Ll210 sublines. The concentration of GSH was increased 5.34 fold in L1210Cis, 2.83 fold in L1210VcR, and 1.78 fold in L1210AdR, compared to L1210. The activities of GCS and GGT were increased in drug-resistant L1210 sublines. The GST activity was increased in L1210VcR and L1210Cis but decreased in L1210AdR compared to Ll210. Expression of GCS, GGT, and GST-${\pi}$ genes were increased in the resistant L1210 sublines compare to the parent L1210 in northern blot analyses. Overexpression of GCS, GGT, and GST-${\pi}$ were observed in the resistant sublines, and the increases of the concentration of glutathione and the activities of GCS and GGT in the resistant sublines may be involved in a part of the drug-resistance in the resistant sublines.

  • PDF

Isolation of Deletion Mutants by Reverse Genetics in Caenorhabditis elegans

  • Park, Byung-Jae;Lee, Jin ll;Lee, Jiyeon;Kim, Sunja;Choi, Kyu Yeong;Park, Chul-Seung;Ahn, Joohong
    • Animal cells and systems
    • /
    • v.5 no.1
    • /
    • pp.65-69
    • /
    • 2001
  • Obtaining mutant animals is important for studying the function of a particular gene. A chemical mutagenesis was first carried out to generate mutations in C. elegans. In this study, we used ultraviolet-activated 4,5',8-trimethylpsoralen to induce small deletion mutations. A library of mutagenized worms was prepared for recovery of candidate animals and stored at $15^{\circ}C$ during screening instead of being made into a frozen stock library. In order to isolate deletion mutations in target genes, a polymerase chain reaction (PCR)-based screening method was used. As a result, two independent mutants with deletions of approximately 1.0 kb and 1.3 kb were isolated. This modified and improved reverse genetic approach was proven to be effective and practical for isolating mutant animals to study gene function at the organismal level.

  • PDF

Production and Characterization of Phenylalanine Ammonia-lyase from Rhodotorula aurantiaca K-505

  • Cho, Dae-Haeng;Chae, Hee-Jeong;Kim, Eui-Yong
    • Preventive Nutrition and Food Science
    • /
    • v.2 no.4
    • /
    • pp.354-359
    • /
    • 1997
  • Optimal cultivation conditions for the production of phenylalanine ammonia-lyase(PAL) from Rhodotorula aurantiaca K-505 were selected, and the kinetic parameters of the produced PAL were determined. The most suitable carbon and nitrogen sources were glucose and tryptone, respectively. The strain expressed PAL constituttively when using the optimized semi-complex media. High cell density culture could be critical for maximal production of PAl since the PAL ynthesis was growth associated. maximum PAL activity was observed at initial pH 6.0. although the ll growth was not markedly affected by temperature between 22 and 28$^{\circ}C$, the cells yielded the maximum PAL activity when cultivated at 22$^{\circ}C$. The maximum activity for deamination of L-phenylalnine to trans-cinnamic acid was observed around pH 8.8. The PAL activity gave the maximum at 45$^{\circ}C$, and greatly decreased at higher than 5$0^{\circ}C$. Activation energy({TEX}$E_{a}${/TEX}) calculated from Arrhenius equation was 6.28 kcal/mol in the range of 22$^{\circ}C$ to 4$0^{\circ}C$. A oolf plot showed that the enzyme reaction follows Michaelis-Menten equation, whose {TEX}$K_{M}${/TEX} and {TEX}$V_{max}${/TEX} values were 4.65$\times${TEX}$10^{-3}${/TEX} M and 0.89$\mu$ mol/mg-min respectively.

  • PDF

The Effects of the Fruits of Foeniculum vulgare on Skin Barrier Function and Hyaluronic Acid Production in HaCaT Keratinocytes (HaCaT 세포에서 회향 열매의 피부장벽기능과 hyaluronic acid 생성에 미치는 영향)

  • Yu, Hak Yin;Yang, In Jun;Lincha, V.R;Park, In Sik;Lee, Dong-Ung;Shin, Heung Mook
    • Journal of Life Science
    • /
    • v.25 no.8
    • /
    • pp.880-888
    • /
    • 2015
  • Foeniculum vulgare (FV) has long been used in traditional medicine for the treatment of inflammatory diseases. In addition, it is usually known as an important medicinal and aromatic plant widely used as a carminative, digestive, lactogogue, and diuretic, and for treating respiratory and gastrointestinal disorders. The skin barrier protects against the invasion of pathogens, fends off chemical and physical assaults, and protects against extensive water loss. In this study, the effects of solvent-fractionated FV fruits on strengthening the skin barrier and maintaining moisture, as well as their antifungal activity, were investigated in human keratinocyte (HaCaT) cells. The expression of involucrin, loricrin, filaggrin, hyaluronic acid synthase, human β defensin, and cathelicidin genes and proteins was measured by reverse transcription polymerase chain reaction (RT-PCR) and western blotting. The production of hyaluronic acid was determined by enzyme-linked immunosorbent assay (ELISA). The butanol fraction increased the expression of involucrin and filaggrin. Both the ethyl acetate and the butanol fractions increased hyaluronic acid production by promoting the expression of hyaluronic acid synthase-1. Although the antimicrobial peptides were increased by FV crude extract and its fractions, the samples did not show a significant effect compared to the normal group. These results suggest that the butanol fraction of FV could be very useful in cosmetics for the treatment of dermatological diseases.

Isolation and Characterization of $A{\alpha}$ mating locus from Schizophyllum commune (치마버섯(Schizophyllum commune)으로부터 $A{\alpha}$ mating locus의 분리 및 특성)

  • Park, Dong-Chul;Novotny, Charles P.;Ullich, Robert C.;Lee, Kap-Duk;Lee, Kap-Rang
    • The Korean Journal of Mycology
    • /
    • v.22 no.3
    • /
    • pp.247-253
    • /
    • 1994
  • This study was carried out to isolate and characterize $A{\alpha}$ mating locus controlling fruiting body formation directly in the Basidiomycete Schzophyllum commune growing in the North America. Total numbers of genomic library of S. commune UVM1-34 was about $2{\times}10^4$ cells. About 90% library was appeared to have about 35 kb inserted genome DNA in cosmid pTC20 vector. 6 clones were proved to have positive signal to probes within Z and Y region in colony and southern hybridization. In the mating activity test, all the 6 positive clones were appeared to have $A{\alpha}3$ mating activity although they had two different restriction patterns. pSC13 containing 5.7 Kb PstI-fragment of UVM 1-34 $A{\alpha}3$ allele showed about 50% clamp cell formation indicating mating activity when cotransformation was done together with cosmid pTC20.

  • PDF

Inhibition of Adipogenesis in 3T3-Ll Adipocytes with Water and Ethanol Extracts of Cudrania tricuspidata Leaves (꾸지뽕잎(Cudrania tricuspidata) 추출물의 3TS-L1 세포 분화 억제)

  • Do, Gun-Pyo;Lee, Hye-Jin;Do, Jeong-Ryong;Kim, Hyun-Ku
    • Food Science and Preservation
    • /
    • v.18 no.2
    • /
    • pp.244-249
    • /
    • 2011
  • The inhibitory effects of adipogenesis in 3T3-L1 adipocytes with water and ethanol extracts of Curdrania tricuspidata leaves were investigated. The lipid accumulation of ethanol extract from Curdrania tricuspidata leaves at a concentration of 5 mg/mL was reduced by 50% compared with control cells, whereas water extracts reduced by 86%. The survival rate of cell viability test showed 80% in the growth of cells at concentrations of 0.5-5 mg/mL. The contents of DNJ(1-deoxynojirimycin) and rutin were higher in leaves than those of stem. Moreover, DNJ contents were detected in large amounts in ethanol extract ($3921.3{\mu}g/g$) compared with water extract ($2906.3{\mu}g/g$). On the contrary, the rutin contents was detected in large amounts in water extracts compared with ethanol extract. These results indicate that Curdrania tricuspidata leaves extract is potential sources of natural antiobesity agent.

Anti-melanogenesis effect of 4-hydroxy-5-methyl-3[2H]-furanone, an antioxidant isolated from pine needles (솔잎에서 분리된 항산화 물질인 4-hydroxy-5methyl-3[2H]-furanone의 멜라닌 생성 억제작용)

  • 부용출;전체옥
    • Journal of the Society of Cosmetic Scientists of Korea
    • /
    • v.20 no.1
    • /
    • pp.1-13
    • /
    • 1994
  • An antioxidative compound was isolated from pine needles. This compound was identified as 4-hydroxy-5-methyl-3[2H]-furanone on the basis of spectroscopic evidences. It scavenged 1,1-diphenyl-2-picrylhydrazyl free radicals more efficiently than maltol and tocopherol did. It exhibited an inhibitory effect on the lipid peroxidation of rat liver microsome induced by Fe(ll)/ascorbate, and the protective effect against UV cytotoxicity in cultured human fibroblasts. In addition, HMF appeared to prevent the cellular melanogenesis in the cultured murine melanoma cells, more effectively than kojic acid, a well known inhibitor of melanogenesis, while the former was not so effective as the latter for the inhibilion of the tyrosinase. Considering that cellular melanogenesis is a metabolic process triggered by oxidative stress, it was tentatively deduced that the antioxidative property of HMF may afford the effect against cellular pigmentation by alleviating the causative stress. This study provided a novel inhibitor of melanogenesis, that might be useful for the cosmetic applications.

  • PDF