• Title/Summary/Keyword: KB cell

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Synergistic Effects of Natural Medicinal Plant Extracts on Growth Inhibition of Carcinoma (KB) Cells under Oxidative Stress (산화적 상해로 인한 상피세포암 세포(KB) 억제에 미치는 천연약용식물 추출물의 상승효과)

  • Kim, Jeong-Hee;Ju, Eun-Mi;Kim, Jin-Kyu
    • Journal of Radiation Protection and Research
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    • v.25 no.4
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    • pp.183-190
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    • 2000
  • Medicinal plants with synergistic effects on growth inhibition of canter cells under oxidative stress were screened in this study. Methanol extracts from 51 natural medicinal plants which were reported to have anticancer effort on hepatoma stomach cancer or colon cancers which are frequently found in Korean, were prepared and screened for their synergistic activity on growth inhibition of cancer cells under chemically-induced oxidative stress by using MTT assay. Twenty seven samples showed synergistic activity on the growth inhibition in various extent under chemically-induced oxidative stress. Among those samples, eleven samples such as Melia azedarach, Agastache rugosa, Catalpa ovata, Prunus persica, Sinomenium acutum Pulsatilla koreana, Oldenlandia diffusa, Anthriscus sylvestris, Schizandra chinensis, Gieditsia sinensis, Cridium officinale, showed decrease in $IC_{50}$ values more than 50%, other 16 samples showed decrease in $IC_{50}$ values between 50-25%, compared with the value acquired when medicinal plant sample was used alone. Among those 11 samples, extract of Catalpa ovata showed the highest activity. $IC_{50}$ values were decrease to 61% and 28% when carcinoma cells were treated with Catalpa ovata extract in combination of 75 and 100 uM of hydrogen peroxide, respectively.

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Design of a Large-density MTP IP (대용량 MTP IP 설계)

  • Kim, YoungHee;Ha, Yoon-Kyu;Jin, Hongzhou;Kim, SuJin;Kim, SeungGuk;Jung, InChul;Ha, PanBong;Park, Seungyeop
    • Journal of IKEEE
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    • v.24 no.1
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    • pp.161-169
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    • 2020
  • In order to reduce the manufacturing cost of MCU chips used in applications such as wireless chargers and USB-C, compared to DP-EEPROM (Double Poly EEPROM), which requires 3 to 5 additional process masks, it is even more necessary MTP(Multi-Time Programmable), which is less than one additional mask and have smaller unit cell size. In addition, in order to improve endurance characteristics and data retention characteristics of the MTP memory cell due to E/P(Erase / Program) cycling, the distribution of the VTP(Program Threshold Voltage) and the VTE(Erase Threshold Voltage) needs to be narrow. In this paper, we proposed a current-type BL S/A(Bit-Line Sense Amplifier) circuit, WM(Write Mask) circuit, BLD(BL Driver) circuit and a algorithm, which can reduce the distribution of program and VT and erase VT, through compare the target current by performing the erase and program pulse of the short pulse several times, and if the current specification is satisfied, the program or erase operation is no longer performed. It was confirmed that the 256Kb MTP memory fabricated in the Magnachip semiconductor 0.13㎛ process operates well on the wafer in accordance with the operation mode.

Inulooligosaccharide Production from Inulin by Saccharomyces cerevisiae Strain Displaying Cell-Surface Endoinulinase

  • Kim Hyun-Chul;Kim Hyun-Jin;Choi Woo-Bong;Nam Soo-Wan
    • Journal of Microbiology and Biotechnology
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    • v.16 no.3
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    • pp.360-367
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    • 2006
  • The endoinulinase gene (inu1) from Pseudomonas mucidolens was expressed on the cell surface of Saccharomyces cerevisiae by fusing with Aga2p linked to the membrane anchored protein, Aga1p. The inu1 gene of P. mucidolens was subcloned into the surface display vector, pCTcon (GAL1 promoter). The constructed plasmid, pCTENIU (8.5kb), was then introduced to S. cerevisiae EBY100 cells and the yeast transformants selected on synthetic defined media lacking uracil and inulin-containing media. The inu1 gene under the control of the GAL1 promoter was successfully expressed in the yeast transformants, and the surface display of endoinulinase confirmed by immunofluorescence microscopy, along with its enzymatic ability to form inulooligosaccharides (IOSs) from inulin. The total endoinulinase activity reached about 2.31 units/ml when the yeast transform ants were cultivated on a YPDG medium. To efficiently hydrolyze the inulin, various reaction conditions were examined, including the pH, temperature, and inulin source. The optimized conditions were then determined as follows: pH, 7.0; temperature, $50^{\circ}C$; inulin source, Jerusalem artichoke. Under the optimized condition and 46 units of endoinulinase per g of inulin, IOSs started to be produced after 10 min of enzymatic reaction. The highest yield, 71.2% of IOSs, was achieved after 30 h of reaction without any significant loss of the initial enzyme activity. As a result of the reaction with inulin, IOSs consisting of inulobiose (F2), inulotriose (F3), inulotetraose (F4), and inulopentaose (F5) were produced, and F4 was the major product.

Comparative Analysis of Cytotoxicity of Fluoride Compounds on Oral Cancer and Osteosarcoma Cells (불소화합물의 골육종 및 구강암 세포주에 대한 독성의 비교분석)

  • 송제선;이백수;김정희
    • Environmental Analysis Health and Toxicology
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    • v.14 no.3
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    • pp.113-120
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    • 1999
  • Fluorination of drinking water has been used world widely to reduce the incidence of caries. Recently, contradictory results on the cytotoxicity of fluoride compounds are reported. In addition, there are attempts to use fluorosilicate for fluorination of drinking water in Korea, therefore, we tried to analyze the cytotoxicity of fluoride compounds on oral epidermoid carcinoma (KB and A253) and osteosarcoma (HOS and MG-63) cells in this study. We treated cells with 0, 10, 50 and 250 ppm of fluorosilicic acid (domestic or from Fluka, F$\_$6/H$_2$Si), sodium fluorosilicate (F$\_$6/Na$_2$Si), sodium fluoroacetate (FCH$_2$CO$_2$Na), sodium fluoride (NaF) or potassium fluoride(KF) and measured the relative cell survival by MTT assay. At the concentration of < 10ppm, no significant cytotoxicity was observed. At 50 ppm, each cells revealed different response to fluoride treatment. Among cells used in this study, MG-63 was the most resistant to fluoride treatment. Comparable toxicity data from domestic and imported fluorosilicic acids were obtained. When we compared the relative cytotoxicity of fluoride compounds against their fluoride contents, the differences in relative cell survival were smaller. Most of cells showed < 20% of survival at 250 ppm. In order to analyze the pH dependence of the cytotoxicity of fluorosilicates, the pH of cell culture media containing fluorosilicate was adjusted to 7.4 or 6.5 and the relative cytotoxicity was measured. At lower pH, about 10% higher cytotoxicity was obtained. Thus, our data suggested that the toxicity of domestic fluorosilicic acid was similar to that of fluorosilicic acid from Fluka, and the cytotoxicity of fluoride compounds was dependent on the relative content of fluoride and pH.

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HOXB5 Directly Regulates the Expression of IL-6 in MCF7 Breast Cancer Cells

  • Kim, Jie Min;Lee, Ji-Yeon;Kim, Myoung Hee
    • Biomedical Science Letters
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    • v.23 no.3
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    • pp.272-276
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    • 2017
  • HOX genes are transcription factors that play important roles in body patterning and cell fate specification during normal development. In previous study, we found aberrant overexpression of HOXB5 in breast cancer tissues and cell lines, and demonstrated that HOXB5 is important in regulation of cell proliferation, tamoxifen resistance, and invasiveness through the epithelial-mesenchymal transition (EMT). Although the relationship between HOXB5 and phenotypic changes in MCF7 breast cancer cells has been studied, the molecular function of HOXB5 as a transcription factor remains unclear. IL-6 has been reported to be involved in not only inflammation but also cancer progression, which is characterized by the increase of growth speed and invasiveness of tumor cells. In this study, we selected Interleukin-6 (IL-6) as HOXB5 putative downstream target gene and discovered that HOXB5 transcriptionally up-regulated the expression of IL-6 in HOXB5 overexpressing MCF7 cells. The upstream region (~1.2 kb) of IL-6 promoter turned out to contain several putative HOX consensus binding sites. Chromatin immunoprecipitation assay confirmed that HOXB5 directly binds to the promoter region of IL-6 and positively regulated the expression of IL-6. These data all together, indicate that HOXB5 promotes IL-6 transcription by actively binding to the putative binding sites located in the upstream region of IL-6, which enable to increase its promoter activity in MCF7 breast cancer cells.

The Secretion Optimization of Oligopeptide with His-Pro Repeats in Bacillus subtilis and Its Anti-Diabetic Effects (고초균에서 His-Pro 반복서열을 갖는 Oligopeptide의 분비 최적화 및 항당뇨 효과)

  • Jeong, Seon Hwa;Choi, Jang Won
    • KSBB Journal
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    • v.32 no.1
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    • pp.71-82
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    • 2017
  • To verify anti-diabetic effect of oligopeptide with His-Pro repeats (mHP peptide), the oligopeptide was first secreted and optimized using the secretion vector, pRBAS with alkaline protease gene promoter and the signal sequence in Bacillus subtilis and directly the anti-diabetic effect of the mHP peptide was investigated in insulinoma cell, RINm5F cell line. The oligopeptide gene was obtained by annealing oligonucleotides with repeated His-Pro sequence and finally was constructed as 18 dipeptides (108 bp and 4.0 kDa) coding gene, named oligopeptide with His-Pro repeats (mHP peptide) to make cyclo(His-Pro) known to be anti-diabetic effects. The region encoding the oligopeptide gene was subcloned into the pRBAS secretion vector (E.coli-Bacillus shuttle vector) after PCR amplification using the designed primers including initiation and termination codons and His tag, named pRBAS-mHP (6.56 kb). To optimize secretion of the oligopeptide, various culture conditions were investigated in Bacillus subtilis LKS. As a result, the secreted oligopeptide was maximally measured (approximately $59.6{\mu}g/mL$) in 3 L batch culture and the highest secretion was achieved at $30^{\circ}C$, PY medium, and carbon sources (particularly barley and glycerol). In the RINm5F cells treated with 2 mM STZ, the oligopeptide treatment (0.1 mg/mL) restored the cell viability (10%) and reduced the nitric oxide (NO) generation (35%) and DNA fragmentation (90%). And also, insulin secretion level was increased to 17% higher than in STZ-treated RINm5F cells. These results suggest that the oligopeptide with His-Pro repeats could be a candidate material for anti-diabetic agent against STZ-induced diabetes.

Inhibitory Effect of Scorpion MeOH Extract on Nitric Oxide and Cytokine Production in Lipopolysaccharide - Activated Raw 264.7 Cells (전갈 메탄올추출물이 LPS로 유도된 Raw 264.7 cell에서의 nitric oxide 및 cytokine에 미치는 영향)

  • Choi, Jun-Hyeok;Lee, Jong-Rok;Jee, Seon-Young;Kim, Sang-Chan
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.3
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    • pp.721-727
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    • 2007
  • Scorpion (SCP) has been clinically used for the treatment of endogenous wind to relieve convulsion, clearing away toxins, resolving hard masses and removing obstruction in the collaterals to relieve pain. Recent studies showed that scorpion toxins that affect the activating mechanism of sodium channels and indian black scorpion venom induced anti-proliferative and apoptogenic activity against human leukemic cell lines U937 and K562. There is lack of studies regarding the effects of SCP on the immunological activities. The present study was conducted to evaluate the effect of SCP on the regulatory effects of cytokines and nitric oxide (NO) for the immunological activities in Raw 264.7 cells. After the treatment of SCP MeOH extract dissolved in media for 1 h prior to the addition of lipopolysaccharide (LPS: 1 ${\mu}$g/ml), cell viability was measured by MTT assay, NO production was monitored by measuring the nitrite content in culture medium. Inducible nitric oxide synthase (iNOS) was determined by immunoblot analysis, and levels of cytokine were analyzed by sandwich immunoassays. As results, SCP inhibited the production of nitrite and nitrate (0.3 and 1.0 mg/ml), iNOS and p-$I_KB_{\alpha}$ protein, tumor necrosis factor-${\alpha}$ (0.3 and 1.0 mg/ml), interleukin-1${\beta}$ (0.3 and 1.0 mg/ml) and interleukin-6 (1.0mg/ml) in Raw 264.7 cells activated with LPS. These findings suggest that SCP can produce anti-inflammatory effect, which may play a role in adjunctive therapy in Gram-negative bacterial infections.

Brief Introduction of Research Progresses in Control and Biocontrol of Clubroot Disease in China

  • He, Yueqiu;Wu, Yixin;He, Pengfei;Li, Xinyu
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.45-46
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    • 2015
  • Clubroot disease of crucifers has occurred since 1957. It has spread to the whole China, especially in the southwest and nourtheast where it causes 30-80% loss in some fields. The disease has being expanded in the recent years as seeds are imported and the floating seedling system practices. For its effective control, the Ministry of Agriculture of China set up a program in 2010 and a research team led by Dr. Yueqiu HE, Yunnan Agricultural University. The team includes 20 main reseachers of 11 universities and 5 institutions. After 5 years, the team has made a lot of progresses in disease occurrence regulation, resources collection, resistance identification and breeding, biological agent exploration, formulation, chemicals evaluation, and control strategy. About 1200 collections of local and commercial crucifers were identified in the field and by artificiall inoculation in the laboratories, 10 resistant cultivars were breeded including 7 Chinese cabbages and 3 cabbages. More than 800 antagostic strains were isolated including bacteria, stretomyces and fungi. Around 100 chemicals were evaluated in the field and greenhouse based on its control effect, among them, 6 showed high control effect, especially fluazinam and cyazofamid could control about 80% the disease. However, fluzinam has negative effect on soil microbes. Clubroot disease could not be controlled by bioagents and chemicals once when the pathogen Plasmodiophora brassicae infected its hosts and set up the parasitic relationship. We found the earlier the pathogent infected its host, the severer the disease was. Therefore, early control was the most effective. For Chinese cabbage, all controlling measures should be taken in the early 30 days because the new infection could not cause severe symptom after 30 days of seeding. For example, a biocontrol agent, Bacillus subtilis Strain XF-1 could control the disease 70%-85% averagely when it mixed with seedling substrate and was drenching 3 times after transplanting, i.e. immediately, 7 days, 14 days. XF-1 has been deeply researched in control mechanisms, its genome, and development and application of biocontrol formulate. It could produce antagonistic protein, enzyme, antibiotics and IAA, which promoted rhizogenesis and growth. Its The genome was sequenced by Illumina/Solexa Genome Analyzer to assembled into 20 scaffolds then the gaps between scaffolds were filled by long fragment PCR amplification to obtain complet genmone with 4,061,186 bp in size. The whole genome was found to have 43.8% GC, 108 tandem repeats with an average of 2.65 copies and 84 transposons. The CDSs were predicted as 3,853 in which 112 CDSs were predicted to secondary metabolite biosynthesis, transport and catabolism. Among those, five NRPS/PKS giant gene clusters being responsible for the biosynthesis of polyketide (pksABCDEFHJLMNRS in size 72.9 kb), surfactin(srfABCD, 26.148 kb, bacilysin(bacABCDE 5.903 kb), bacillibactin(dhbABCEF, 11.774 kb) and fengycin(ppsABCDE, 37.799 kb) have high homolgous to fuction confirmed biosynthesis gene in other strain. Moreover, there are many of key regulatory genes for secondary metabolites from XF-1, such as comABPQKX Z, degQ, sfp, yczE, degU, ycxABCD and ywfG. were also predicted. Therefore, XF-1 has potential of biosynthesis for secondary metabolites surfactin, fengycin, bacillibactin, bacilysin and Bacillaene. Thirty two compounds were detected from cell extracts of XF-1 by MALDI-TOF-MS, including one Macrolactin (m/z 441.06), two fusaricidin (m/z 850.493 and 968.515), one circulocin (m/z 852.509), nine surfactin (m/z 1044.656~1102.652), five iturin (m/z 1096.631~1150.57) and forty fengycin (m/z 1449.79~1543.805). The top three compositions types (contening 56.67% of total extract) are surfactin, iturin and fengycin, in which the most abundant is the surfactin type composition 30.37% of total extract and in second place is the fengycin with 23.28% content with rich diversity of chemical structure, and the smallest one is the iturin with 3.02% content. Moreover, the same main compositions were detected in Bacillus sp.355 which is also a good effects biocontol bacterial for controlling the clubroot of crucifer. Wherefore those compounds surfactin, iturin and fengycin maybe the main active compositions of XF-1 against P. brassicae. Twenty one fengycin type compounds were evaluate by LC-ESI-MS/MS with antifungal activities, including fengycin A $C_{16{\sim}C19}$, fengycin B $C_{14{\sim}C17}$, fengycin C $C_{15{\sim}C18}$, fengycin D $C_{15{\sim}C18}$ and fengycin S $C_{15{\sim}C18}$. Furthermore, one novel compound was identified as Dehydroxyfengycin $C_{17}$ according its MS, 1D and 2D NMR spectral data, which molecular weight is 1488.8480 Da and formula $C_{75}H_{116}N_{12}O_{19}$. The fengycin type compounds (FTCPs $250{\mu}g/mL$) were used to treat the resting spores of P. brassicae ($10^7/mL$) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be lyzed by the FTCPs of XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol. In the five selected medium MOLP, PSA, LB, Landy and LD, the most suitable for growth of strain medium is MOLP, and the least for strains longevity is the Landy sucrose medium. However, the lipopeptide highest yield is in Landy sucrose medium. The lipopeptides in five medium were analyzed with HPLC, and the results showed that lipopeptides component were same, while their contents from B. subtilis XF-1 fermented in five medium were different. We found that it is the lipopeptides content but ingredients of XF-1 could be impacted by medium and lacking of nutrition seems promoting lipopeptides secretion from XF-1. The volatile components with inhibition fungal Cylindrocarpon spp. activity which were collect in sealed vesel were detected with metheds of HS-SPME-GC-MS in eight biocontrol Bacillus species and four positive mutant strains of XF-1 mutagenized with chemical mutagens, respectively. They have same main volatile components including pyrazine, aldehydes, oxazolidinone and sulfide which are composed of 91.62% in XF-1, in which, the most abundant is the pyrazine type composition with 47.03%, and in second place is the aldehydes with 23.84%, and the third place is oxazolidinone with 15.68%, and the smallest ones is the sulfide with 5.07%.

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MAKING IN VIVO MODEL TO STUDY ABOUT HUMAN ORAL CANCER (I) (구강암 연구를 위한 동물실험모델의 개발(I))

  • Park, Hyung-Kook;Kim, Yong-Kack
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.19 no.3
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    • pp.300-310
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    • 1997
  • In order to make in vivo model of human oral squamous cell cancer, we brought up BALB/C nude mice in specially designed housystem, and maintained some kind of human oral squamous cancer cell lines ; KB, SCC-4, SCC-9, SCC-15, SCC-25. Various concentration of cancer cells were inoculated subcutaneouly into flank area of nude mice. We observed each nude mouse more than 5 weeks after tumor inoculation. We appraised the results, measured the tumor size, and calculated the growing tumor volumes after tumor inoculation according to cancer cell line and concentration of cancer cells in media. Some cancer cell lines were rapidly growing in nude mice, but some cancer cell line couldn't grow in nude mice and resorbed completely. And in some cancer cell line, some nude mice showed continuously growing tumor, but other didn't show any tumor growing. And as a new try, we implanted specially disigned caps on the back of nude mice, and cancer cell lines were brought into the caps with media. We removed the cap after 1 week, and observed over 4 weeks. The shape and size of growing tumor were observed.

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Expression of Yippee-Like 5 (YPEL5) Gene During Activation of Human Peripheral T Lymphocytes by Immobilized Anti-CD3 (인체 말초혈액의 활성화 과정 중 yippee-like 5 (YPEL5) 유전자의 발현 양상)

  • Jun, Do-Youn;Park, Hye-Won;Kim, Young-Ho
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1641-1648
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    • 2007
  • Yippee-like proteins, which have been identified as the homolog of Drosophila yippee protein containing a zinc-finger domain, are known to be highly conserved among eukaryotes. However, their functional roles are still poorly understood. Recently we initiated ordered differential display (ODD)-polymerase chain reaction (PCR) to isolate genes of which expressions are altered following activation of human T cells. On the ODD-PCR image, one PCR-product detected in unstimulated T cells was not detectable at the time when the activated T cells traversed near $G_1/S$ boundary following activation by immobilized anti-CD3. Cloning and nucleotide sequence analysis revealed that the PCR-product was yippee-like 5 (YPEL5) gene, which was known as a human homolog of the Drosophila yippee gene. Northern blot analysis confirmed the amount of ${\sim}2.2$ kb YPEL5 mRNA expression detectable in unstimulated T cells was sustained until 1.5 hr after activation and then rapidly declined to undetectable level by 5 hr. Ectopic expression of YPEL5 gene in human cervix epitheloid carcinoma HeLa cells caused a significant reduction in cell proliferation to the level of 47% of the control. Expression of GFP-YPEL5 fusion protein in HeLa cells showed its nuclear localization. These results demonstrated that the expression level of human YPEL5 mRNA was negatively regulated in the early stage of T cell activation, and suggested that YPEL5 might exert an inhibitory effect on the cell proliferation as a nuclear protein.