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Apoptotic Cell Death by Melittin through Induction of Bax and Activation of Caspase Proteases in Human Lung Carcinoma Cells (Bax의 발현증가 및 Caspase의 활성을 통한 봉독약침액 Melittin의 인체폐암세포 Apoptosis 유발에 관한 연구)

  • Ahn, Chang-beohm;Im, Chun-woo;Kim, Cheol-hong;Youn, Hyoun-min;Jang, Kyung-jeon;Song, Choon-ho;Choi, Yung-hyun
    • Journal of Acupuncture Research
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    • v.21 no.2
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    • pp.41-55
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    • 2004
  • Objective : To investigate the possible molecular mechanism (s) of melittin as a candidate of anti-cancer drug, we examined the effects of the compound on the growth of human lung carcinoma cell line A549. Methods : Growth inhibitory study, flow cytometry analysis, SDS-polyacrylamide gel electrophoresis and Western blot analysis, RT-PCR and in vitro caspases activity assay were performed. Results : Melittin treatment declined the cell viability of A549 cells in a concentration-dependent manner, which was associated with induction of apoptotic cell death. Melittin treatment down-regulated the levels of Bcl-XS/L mRNA and protein expression of A549 cells, an anti-apoptotic gene, however, the those of Bax, a pro-apoptotic gene, were up-regulated. Melittin induced the proteolytic cleavage and activation of caspase-3 and caspase-9 protease in a dose-dependent manner without alteration of inhibitor of apoptosis proteins family and Akt expression. Western blot analysis and RT-PCR data revealed that the levels of tumor suppressor p53 and cyclin-dependent kinase inhibitor p21 were also remained unchanged. Conclusions : Taken together, these findings suggest that melittin-induced inhibition of human lung cancer cell growth is associated with the induction of apoptotic cell death via regulation of several major growth regulatory gene products, and melittin may have therapeutic potential in human lung cancer.

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Effects of Hyperbaric Pressure on Cellular Morphology, Proliferation and Protein Expression of Jurkat Cell

  • Oh, Eun-Ha;Oh, Sang-Nam;Im, Ho-Sub;Lee, Joo-Hyun;Kim, Jin-Young;Moon, Joo-Hee;Hong, Eun-Young;Kim, Yang-Hee;Yang, Min-Ho;Lim, Yong-Chul;Park, Sun-Young;Lee, Eun-Il;Sul, Dong-Geun
    • Molecular & Cellular Toxicology
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    • v.1 no.2
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    • pp.116-123
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    • 2005
  • The application of high pressure on cellular morphology, proliferation and protein expression of Jurkat cells (human T lymphocyte cell line) has been extensively investigated. In the present study, we manufactured a novel pressure chamber that modulates 5% $CO_{2}$, temperature and pressure (up to 3 ATA). Jurkat cells was incubated 2 ATA pressure and analyzed cellular morphology and growth using an electron microscopy and MTT assay. The cells showed the morphological changes in the cell surface, which appeared to cause a severe damage in cell membrane. The growth rate of the cells under 2 ATA pressure decreased as cultured time got increased. Furthermore, a long term exposure of high pressure on Jurkat cells may act as one of the important cellular stresses that leads to inducing cell death. Cellular proteomes were separated by 2-dimensional electrophoresis with pH 3-10 ranges of IPG Dry strips. And many proteins showed significant up-and-down expressions with hyperbaric pressure. Out of all, 10 spots were identified significantly using matrix-assisted laser desorption/ionization-time of fight (MALDI-TOF) mass spectrometry. We and found that 9 protein expressions were decreased and one protein, heat shock protein HSP 60, was increased in Jurkat cells under 2 ATA. Identified proteins were related to lipid metabolism and signal transduction.

Gamma Knife Radiosurgery for Ten or More Brain Metastases

  • Kim, Chang-Hyun;Im, Yong-Seok;Nam, Do-Hyun;Park, Kwan;Kim, Jong-Hyun;Lee, Jung-Il
    • Journal of Korean Neurosurgical Society
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    • v.44 no.6
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    • pp.358-363
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    • 2008
  • Objective : This study was performed to assess the efficacy of GKS in patients with ten or more brain metastases. Methods : From Aug 2002 to Dec 2007, twenty-six patients (13 men and 13 women) with ten or more cerebral metastatic lesions underwent GKS. The mean age was 55 years (32-80). All patients had Karnofsky performance status (KPS) score of 70 or better. According to recursive partitioning analysis (RPA) classification, 3 patients belonged to class I and 23 to class II. The location of primary tumor was lung (21), breast (3) and unknown (2). The mean number of the lesions per patient was 16.6 (10-37). The mean cumulated volume was 10.9 cc (1.0-42.2). The median marginal dose was 15 Gy (9-23). Overall survival and the prognostic factors for the survival were retrospectively analyzed by using Kaplan Meier method and univariate analysis. Results : Overall median survival from GKS was 34 weeks (8-199). Local control was possible for 79.5% of the lesions and control of all the lesions was possible in at least 14 patients (53.8%) until 6 months after GKS. New lesions appeared in 7 (26.9%) patients during the same period. At the last follow-up, 18 patients died; 6 (33.3%) from systemic causes, 10 (55.6%) from neurological causes, and 2 (11.1 %) from unknown causes. Synchronous onset in non-small cell lung cancer (p=0.007), high KPS score (${\geq}80$, p=0.029), and controlled primary disease (p=0.020) were favorable prognostic factors in univariate analysis. Conclusion : In carefully selected patients, GKS may be a treatment option for ten or more brain metastases.

Antioxidant and Anti-inflammatory Activities of Ethanol Extract from Leaves of Cirsium japonicum

  • Lee, Je-Hyuk;Choi, Soo-Im;Lee, Yong-Soo;Kim, Gun-Hee
    • Food Science and Biotechnology
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    • v.17 no.1
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    • pp.38-45
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    • 2008
  • Antioxidant and anti-rheumatoid activities of Cirsium japonicum leaf extract (CJLE) were investigated in this study. CJLE had similar DPPH radical scavenging activity and reducing power to ascorbic acid and several flavonoids. Rheumatoid arthritis (RA) is a chronic inflammatory tissue-destructive disease, partly related with functions of hyaluronidases (HAases) and collgenases. CJLE ($1,000\;{\mu}g/mL$) had approximately 60.7 and 31.9% inhibition of HAase and collagenase activity, respectively. Also, CJLE inhibited lipopolysaccharide (LPS)-induced nitrite production in a dose-dependent manner, and CJLE ($1,000\;{\mu}g/mL$) suppressed approximately 70% of LPS-induced nitrite production effectively in RAW 264.7 macrophage cells. CJLE had inhibitory effects on the adherence of monocytic THP-1 to human umbilical vein endothelial cell (HUVEC) monolayers to the basal level. Inhibitory effect of CJLE on the adhesion was caused by suppression of tumor necrosis factor-a-upregulated expression of vascular cellular adhesion molecule-1 (VCAM-1) and E-selectin. We expect that CJLE may alleviate the inflammatory process in rheumatoid synovium, and these findings will raise the possibility of the usage of C. japonicum as a traditional pharmaceutical of anti-rheumatoid arthritis.

In vitro cytotoxicity of Acanthamoeba spp. isolated from contact lens containers in Korea by crystal violet staining and LDH release assay

  • Shin, Ho-Joon;Cho, Myung-Soo;Jung, Suk-Yul;Kim, Hyung-Il;Im, Kyung-Il
    • Parasites, Hosts and Diseases
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    • v.38 no.2
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    • pp.99-102
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    • 2000
  • In order to observe the cytotoxicity of Acanthamoeba spp., which were isolated from contact lens containers as ethiological agents for the probable amoebic keratitis in Korea, the crystal violet staining method and LDH release assay were carried out. In the crystal violet staining method, among eight contact lens container isolates, isolate 3 (Acanthauloeba KA/LS5) showed 83.6% and 81.8% of cytotoxicity, and isolate 7 (Acanthamoeba KA/LS37) showed 28.2% and 25.1% of cytotoxicity, in 1 mg/ml and 0.5 mg/ml Iysate treatments, respectively. Acanthamoeba cutbertsoni and A. healyi showed 84.0% and 82.8% of cytotoxicity. Similar results were observed in A. costellunii and A. hafchefti which showed 83.6% and 75.5% or cytotoxicity. Acanthamoeba roureba and A. polyphaga showed 9.0% and 1.7% of cytotoxicity. In the LDH release assay, isolate 3 (20.4%) showed higher cytotoxicity than other isolates in 1 mg/ml Iysate treatment. The results provide that at least isolate 3 has the cytotoxic effect against CHO cells and seems to be the pathogenic strain.

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Cytotoxic Activity and Structure Activity Relationship of Ceramide Analogues in Caki-2 and HL-60 Cells

  • Kim, Yong-Jin;Kim, Eun-Ae;Sohn, Uy-Dong;Yim, Chul-Bu;Im, Chae-Uk
    • The Korean Journal of Physiology and Pharmacology
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    • v.14 no.6
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    • pp.441-447
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    • 2010
  • B13, a ceramide analogue, is a ceramidase inhibitor and induces apoptosis to give potent anticancer activity. A series of thiourea B13 analogues was evaluated for their in vitro cytotoxic activities against human renal cancer Caki-2 and leukemic cancer HL-60 in the MTT assay. Some compounds (12, 15, and 16) showed stronger cytotoxicity than B13 and C6-ceramide against both tumor cell lines, and compound (12) gave the most potent activity with $IC_{50}$ values of 36 and $9\;{\mu}M$, respectively. Molecular modeling of thiourea B13 analogues was carried out by comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA). We obtained highly reliable and predictive CoMSIA models with cross-validated $q^2$ values of 0.707 and 0.753 and CoMSIA contour maps to show the structural requirements for potent activity. These data suggest that the amide group of B13 could be replaced by thiourea, that the stereochemistry of 1,3-propandiol may not be essential for activity and that long alkyl chains increase cytotoxicity.

A Study on the Data Transmission line of communication system (통신시스템의 데이터 전송선로에 대한 연구)

  • Kim Soke-Hwan;Lee Kyeu-jung;Hur Chang-wu
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.9 no.6
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    • pp.1277-1281
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    • 2005
  • FPGA has been widely used in communication system. In this paper, we made 10 layers PCB on protection of signal noise and data lose with FPGA. We analyzed about change of the data transmission speed and length according to input frequency. The length of transmission line from FPGA's output-pin to output-port on PCB board is 13cm and extended lengths for test are 30cm, 60cm and 10cm. We knew that data can be stably transmitted to 100Mbps at transmission line length of 30cm.

대기압에서 발생시킨 헬륨 플라즈마에서의 산소함유량 증가에 따른 폐암세포의 세포내 활성 산소 종 및 세포주기 변화

  • Jo, Hye-Min;Kim, Seon-Ja;Jeong, Tae-Hun;Im, Seon-Hui
    • Proceedings of the Korean Vacuum Society Conference
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    • 2014.02a
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    • pp.261.2-261.2
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    • 2014
  • 저온에서 작동하는 대기압 플라즈마 젯은 생체 조직에의 플라즈마 처리를 가능하게 한다. 이에 이온과 전자, 활성 종, 전기장, UV 등을 발생시키는 플라즈마를 암세포에 처리하여 그에 따른 변화를 관찰하였다. 모세관 타입의 젯에 산소를 반응기체로 흘려주어 헬륨 내 산소 함유량에 따른 활성 산소종의 생성을 확인하였다. 대기압 플라즈마에 의해 생성되는 활성 산소 종(OH, O, electronically excited O (1D), O2 ($1{\Delta}g$) 등)이 세포에 산화 스트레스를 유발할 것이라 예상되어 인체의 폐암 세포[Human lung cancer cell, A549]에 펄스파의 헬륨-산소 플라즈마를 처리한 후, 세포 내 활성 산소 종의 증가량을 비교하였다. 그 결과 적은 양의 산소를 추가하였을 때 세포 내 활성 산소 종의 농도가 증가되었다. 이때 플라즈마에서 발생되는 활성 산소 종(Reactive Oxygen Species, ROS)들은 광 방출 스펙트럼(Optical Emission Spectroscopy)로 확인하였고, 세포내 활성 산소 종은 DCF-DA 염색을 통하여 분석하였다. 이러한 헬륨-산소 플라즈마가 세포 성장의 어떠한 시기에 영향을 미치는지를 알아보기 위하여 세포주기 변화를 분석한 결과, 플라즈마 처리 9시간 후부터 G2/M 주기에 머물러 있음을 확인하였다.

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Melanin Production Inhibitory Activity of the Dendropanax morbifera Leaf Extract Fermented by Lactobacillus plantarum (Lactobacillus plantarum으로 발효시킨 황칠나무 잎 추출물의 피부 미백 관련 효과)

  • Im, Do youn;Lee, Kyoung in
    • Korean Journal of Pharmacognosy
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    • v.47 no.1
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    • pp.18-23
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    • 2016
  • This study was conducted to investigate the tyrosinase inhibitory and melanin production inhibitory activity of the distilled water extract of Dendropanax morbifera leaf (DMW) and the fermented extract by Lactobacillus plantarum (DMF). DMF was prepared by inoculation of L. plantarum after the extraction procedure with distilled water. Fermentation for 48 hours at $37^{\circ}C$ is the most effective condition in this study. In DPPH radical scavenging ability, $SC_{50}$ values of the fermented DMF was $37.9{\mu}g/ml$ as a result of more effective than DMW extract ($52.6{\mu}g/ml$). Moreover, tyrosinase inhibitory activity of DMF showed higher activity than DMW. In nontoxic concentration range, DMF showed strong melanin production inhibitory effect in ${\alpha}$-melanocyte stimulating hormone-stimulated B16F10 cell. As a result, the fermentation of the distilled water extract of D. morbifera leaf by L. plantarum could be applicable to functional materials production for skin-whitening agents.

Crystallization and Preliminary X-Ray Crystallographic Analysis of PAS Factor from Vibrio vulnificus

  • Lee, Jun Hyuck;Kim, Soo Young;Rho, Seong-Hwan;Im, Young Jun;Kim, Young Ran;Kim, Mun-Kyoung;Kang, Gil Bu;Rhee, Joon Haeng;Eom, Soo Hyun
    • Molecules and Cells
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    • v.20 no.3
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    • pp.361-363
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    • 2005
  • Plasmid Achromobacter secretion (PAS) factor is a putative secretion factor that induces the secretion of periplasmic proteins. PAS factor from Vibrio vulnificus was crystallized at 294 K by the hanging drop vapor-diffusion method. It was isolated as a monomer during the purification procedures. The native crystal belongs to the F222 space group with unit cell parameters a = 56.1, b = 74.4, $c=80.0{\AA}$, ${\alpha}={\beta}={\gamma}=90^{\circ}$. The crystal was soaked in cryoprotectant containing 1 M NaBr for 1 h for MAD phasing. The diffraction limit of the Br-MAD data set was $1.9{\AA}$ using synchrotron X-ray irradiation at beam line BL-18B at the Photon Factory, Japan.