• Title/Summary/Keyword: HL-60

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Effect of Electromagnetic Fields on Growth of Human Cell Lines

  • Oh, Se-Jong;Lee, Mi-Kyung;Lee, Seo-Ho;Lee, Jin-Ha;Kim, Dai-Jong;Park, Young-Shik;Lee, Hyeon-Yong
    • Journal of Microbiology and Biotechnology
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    • v.11 no.5
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    • pp.749-755
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    • 2001
  • High Electromagnetic Field (EMF) with an intensity of 1 mT (Tesla) inhibited the growth of both human normal lung and immune T cell down to $20-30\%$, compared to that of an unexposed case. The human T-cells, Jurkat, were more severely affected by EMF than the human lung cells, which showed a relatively slow cell growth and substantial releas of $Ca^+2$ (3.5 times higher than the human T-cells). However, the growth of hepatoma carcinoma, Hep3B, was enhanced by twice that of an unexposed case. The EMF intensity and exposure time did not affect the growth of the cancer cells very much, while it significantly affected the growth of normal cells. Accordingly, it is possible that EMFs may play a role in the initiation of cancer. The EMFs disturbed the signal transduction and membrane systems, such that a five times higher amount of PKC-${\alpha}$ was released from the cell membrane than in the control. Extended exposure to EMFs, for more than 48 hours, also led to 1 $90\%$ necrotic death pattern from apoptotic cell death. Finally, EMF at an intensity of 1mT with a 24-T exposure promoted the differentiation of HL-60 cells to monocytes/macrophages, possibly causing potential acute leukemia.

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Effect of small Black Soybean Fraction on the T cell-mediated Immune Responses in vivo and Proliferation of Leukemia Cells in vitro

  • Oh, Chang-Ho;Shin, Tae-Yong;Chae, Byeong-Suk;Lee, Kyu-Hee;Kim, Ju-Sin;Moon, Mi-Kyeong;Cho, Moon-Gu;Kim, Jong-Hwa;Oh, Suk-Heung;Lee, Tae-Kyoo;Kim, Dae-Keun
    • Natural Product Sciences
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    • v.13 no.2
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    • pp.123-127
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    • 2007
  • We investigated effect of small black soybean fraction (SBSF) T cell-mediated responses for tumor surveillance and proliferation in leukemia cells in vitro. Each SBSF butanol fraction (SBSFBu) and SBSF chloroform fraction (SBSFCh) was administered p.o. once a day far 21 days in BALB/c mice and then levels of serum cytokines and subpopulation of lymphocytes were measured. Moreover, SBSF fraction was treated into the cultured various cell lines for proliferation in leukemia cell lines, NO production by RAW264.7 cells, and expression of p53 gene in U937 leukemia cells. These results showed that SBSFBu increased levels of serum IL-4but not IL-2 and IFN-${\gamma}$, and increased expression of CD4$^+$ T cells and CD8$^+$ T cells in splenocytes in vivo, while SBSFCh increased levels of serum IL-2 and IFN-${\gamma}$ but decreased IL-4, and increased CD8$^+$ T cells but not CD4$^+$ T cells. Moreover, both of SBSFBu and SBSFCh inhibited proliferation of HL60, U937, and L1210 leukemia cell lines in a dose-dependent manner, up-regulated NO production by RAW264.7 cells in a dose-dependent manner, and enhanced expression of p53 gene in U937 leukemia cells. Our findings indicate that SBSFBu and SBSFCh may enhance T cell-dependent immune responses, and that both of SBSFBu and SBSFCh may inhibit proliferation of leukemia cells by up-regulation of NO production and expression of p53 gene.

Immunohistochemical Detection of N-myc Gene Product by Using Antiserum Against Synthetic Peptide (항-펩타이드 항체를 이용한 암유전자 N-myc 산물의 면역조직화학적 검출)

  • Lee, Hyun-Chul;Lee, Wan-Joo;Ahn, Tai-Hew
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.2
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    • pp.167-174
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    • 1987
  • N-myc, a DNA sequence related to the oncogene c-myc, was found to be amplified in untreated primary neuroblastomas and the amplification appeared to be associated with advanced disease at diagnosis and rapid tumor progression. Synthetic peptides have been useful immunogens for generating antisera and monoclonal antibodies to a number of native proteins. In order to identify myc-related protein in the tumor cells, an antiserum against a synthetic hexapeptide (-Glu-Asp-Ile-Trp-Lys-Lys-), whose sequence corresponds to a part of the exon 2 of oncogene N-myc, was prepared by immunizing a rabbit with BSA-conjugated peptide. After ammonium sulfate precipitation and affinity column chromatography, it appeared to be specific to the peptide. Strong nuclear staining in immunoperoxidase method using this serum was observed in both human promyeloid leukemic cell line, HL-60(containing high c-myc copy number), and human neuroblastoma cell line, LA-N-5 (containing high N-myc copy number), whereas LA351 (human lymphoid cell line) cells did not react with the serum. This reaction was completely abrogated by incubating the antiserum with soluble excess peptide. These data suggest that the protein encoded by N-myc could be localized in the nucleus as c-myc protein and this antiserum can be used to detect myc-related tumor cells in clinical samples and to determine if the N-myc expression correlates with genomic amplification in cell lines, untreated primary tumors, and untreated metastases.

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Validation of a Multiplexed Opsonophagocytic Assay for 11 Additional Pneumococcal Serotypes and Its Application to Functional Antibody Evaluation Induced by Pneumococcal Polysaccharide Vaccine

  • Cha, Jihei;Kim, Han Wool;Lee, Ji Hyen;Lee, Soyoung;Kim, Kyung-Hyo
    • Journal of Korean Medical Science
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    • v.33 no.51
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    • pp.340.1-340.14
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    • 2018
  • Background: Various pneumococcal vaccines have been evaluated for immunogenicity by opsonophagocytic assay (OPA). A multiplexed OPA (MOPA) for 13 pneumococcal serotypes was developed by Nahm and Burton, and expanded to 26 serotypes in 2012. The development of new conjugate vaccines with increased valence has necessitated expanded MOPAs to include these additional serotypes. In this study, we validated this expanded MOPA platform and applied to measure antibodies against 11 additional serotypes (2, 8, 9N, 10A, 11A, 12F, 15B, 17F, 20B, 22F, and 33F) in human sera. Methods: All materials, including serum, complement, bacterial master stocks, and HL-60 cells, were evaluated for assay optimization. Following optimization, the assay was validated for accuracy, specificity, and intra- and inter-assay precision with sera from adult donors following standard protocols. The assay was applied to evaluate functional antibodies of 42 sera immunized with 23-valent pneumococcal polysaccharide vaccine (PPV23). Results: The expanded MOPA platform was specific for all serotypes, with the exception of serotype 20. The assay results were highly correlated with those obtained from single-serotype OPA, indicating acceptable accuracy. The coefficients of variation were 7%-24% and 13%-39% in tests of intra- and inter-assay precision, respectively, using three quality-control samples. A MOPA that included 11 additional serotypes in the PPV23 was established and validated with respect to accuracy, specificity, and precision. The opsonic indices of immune sera were obtained using this validated assay. Conclusion: The expanded MOPA will be useful for evaluation of the immunogenicity of PPV23 and future conjugate vaccine formulations.

SUV39H1 is a New Client Protein of Hsp90 Degradated by Chaetocin as a Novel C-Terminal Inhibitor of Hsp90

  • Lian, Bin;Lin, Qian;Tang, Wei;Qi, Xin;Li, Jing
    • Biomolecules & Therapeutics
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    • v.29 no.1
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    • pp.73-82
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    • 2021
  • Hsp90 is often overexpressed with activated form in cancer cells, and many key cellular proteins are dependent upon the Hsp90 machinery (these proteins are called "client protein"). Nowadays, more client proteins and more inhibitors of Hsp90 are being discovered. Chaetocin has been identified as an inhibitor of histone methyl transferase SUV39H1. Herein, we find that Chaetocin is an inhibitor of Hsp90 which binds to the C-terminal of Hsp90α. Chaetocin inhibited a variety of Hsp90 client proteins including AMl1-ETO and BCL-ABL, the mutant fusion-protein in the K562 and HL-60 cells. SUV39H1 mediates epigenetic events in the pathophysiology of hematopoietic disorders. We found that inhibition of Hsp90 by Chaetocin and 17-AAG had ability to induce degradation of SUV39H1 through proteasome pathway. In addition, SUV39H1 interacted with Hsp90 through co-chaperone HOP. These results suggest that SUV39H1 belongs to a client protein of Hsp90. Moreover, Chaetocin was able to induce cell differentiation in the two cells in the concentration range of Hsp90 inhibition. Altogether, our results demonstrate that SUV39H1 is a new client protein of Hsp90 degradated by Chaetocin as a novel C-terminal inhibitor of Hsp90. The study establishes a new relationship of Chaetocin and SUV39H1, and paves an avenue for exploring a new strategy to target SUV39H1 by inhibition of Hsp90 in leukemia.

Test-Retest Reliability of Level-Specific CE-Chirp Auditory Brainstem Response in Normal-Hearing Adults

  • Jamal, Fatin Nabilah;Dzulkarnain, Ahmad Aidil Arafat;Shahrudin, Fatin Amira;Marzuki, Muhammad Nasrullah
    • Journal of Audiology & Otology
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    • v.25 no.1
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    • pp.14-21
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    • 2021
  • Background and Objectives: There is growing interest in the use of the Level-specific (LS) CE-Chirp® stimulus in auditory brainstem response (ABR) due to its ability to produce prominent ABR waves with robust amplitudes. There are no known studies that investigate the test-retest reliability of the ABR to the LS CE-Chirp® stimulus. The present study aims to investigate the test-retest reliability of the ABR to the LS CE-Chirp® stimulus and compare its reliability with the ABR to standard click stimulus at multiple intensity levels in normal-hearing adults. Subjects and Methods: Eleven normal-hearing adults participated. The ABR test was repeated twice in the same clinical session and conducted again in another session. The ABR was acquired using both the click and LS CE-Chirp® stimuli at 4 presentation levels (80, 60, 40, and 20 dBnHL). Only the right ear was tested using the ipsilateral electrode montage. The reliability of the ABR findings (amplitudes and latencies) to the click and LS CE-Chirp® stimuli within the same clinical session and between the two clinical sessions was calculated using an intra-class correlation coefficient analysis (ICC). Results: The results showed a significant correlation of the ABR findings (amplitude and latencies) to both stimuli within the same session and between the clinical sessions. The ICC values ranged from moderate to excellent. Conclusions: The ABR results from both the LS CE-Chirp® and click stimuli were consistent and reliable over the two clinical sessions suggesting that both stimuli can be used for neurological diagnoses with the same reliability.

Test-Retest Reliability of Level-Specific CE-Chirp Auditory Brainstem Response in Normal-Hearing Adults

  • Jamal, Fatin Nabilah;Dzulkarnain, Ahmad Aidil Arafat;Shahrudin, Fatin Amira;Marzuki, Muhammad Nasrullah
    • Korean Journal of Audiology
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    • v.25 no.1
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    • pp.14-21
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    • 2021
  • Background and Objectives: There is growing interest in the use of the Level-specific (LS) CE-Chirp® stimulus in auditory brainstem response (ABR) due to its ability to produce prominent ABR waves with robust amplitudes. There are no known studies that investigate the test-retest reliability of the ABR to the LS CE-Chirp® stimulus. The present study aims to investigate the test-retest reliability of the ABR to the LS CE-Chirp® stimulus and compare its reliability with the ABR to standard click stimulus at multiple intensity levels in normal-hearing adults. Subjects and Methods: Eleven normal-hearing adults participated. The ABR test was repeated twice in the same clinical session and conducted again in another session. The ABR was acquired using both the click and LS CE-Chirp® stimuli at 4 presentation levels (80, 60, 40, and 20 dBnHL). Only the right ear was tested using the ipsilateral electrode montage. The reliability of the ABR findings (amplitudes and latencies) to the click and LS CE-Chirp® stimuli within the same clinical session and between the two clinical sessions was calculated using an intra-class correlation coefficient analysis (ICC). Results: The results showed a significant correlation of the ABR findings (amplitude and latencies) to both stimuli within the same session and between the clinical sessions. The ICC values ranged from moderate to excellent. Conclusions: The ABR results from both the LS CE-Chirp® and click stimuli were consistent and reliable over the two clinical sessions suggesting that both stimuli can be used for neurological diagnoses with the same reliability.

Apoptosis Induction of HCT-15 Cells by Extracts of Undaria pinnatifida with Fermented Micro-organism (미역 발효추출물의 HCT-15 대장암 세포 사멸 유도 효과)

  • Kim, Tae-Yun;Han, Hyo-Sang;Lee, Young-Jong
    • The Korea Journal of Herbology
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    • v.28 no.4
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    • pp.33-40
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    • 2013
  • Objectives : To study the apoptosis effects of fermented Undaria pinnatifida extracts(FUP) against HCT-15 colon cancer cells. Method : By measuring cell proliferation, DNA fragmentation, cell cycle, morphology, and western blot from FUP, the study investigated the effects of the extractions had upon the HCT-15 colon cancer cells, and concluded that the inhibiting effects upon cells were induced by apoptosis. Result : FUP effectively inhibited the growth of HCT-15 colon cancer cells. After analyzing the DNA fragmentation, the study observed a DNA ladder, while examining the cells, and found an increase of sub-G1 hypodiploid cells. On the changes regarding the nucleus of the cells, a condensation of cells and chromatin, as well as an apoptotic body was clearly observed. By observing through western blot from FUP, the study found a decreased level of Bcl-2 from HCT-15 colon cancer cells, but the increased level of Bax and cleaved caspase-3, which as a result induced apoptosis, inhibiting the growth of HCT-15 colon cancer cells. FUP increased the natural death of HCT-15 colon cancer cells by the induction of apoptosis. FUP seemed to have no suppressing effect upon HL-60/MX2 cells. However, compared to the fucoidan, the study was able to clearly observe morphological changes of HCT-15 cells apoptosis, in a 1/2 concentration. Conclusion : FUP had antiproliferative effects on different kinds of cancer cells, while proving especially efficacious against colon cancer cells.

Isolation and Cytotoxic Potency of Endophytic Fungi Associated with Dysosma difformis, a Study for the Novel Resources of Podophyllotoxin

  • Hoa Thi Tran;Giang Thu Nguyen;Hong Ha Thi Nguyen;Huyen Thi Tran;Quang Hong Tran;Quang Ho Tran;Ngoc Thi Ninh;Phat Tien Do;Ha Hoang Chu;Ngoc Bich Pham
    • Mycobiology
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    • v.50 no.5
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    • pp.389-398
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    • 2022
  • Endophytic fungi are promising sources for the production of podophyllotoxin-an important anticancer compound, replacing depleted medical plants. In this study, the endophytes associated with Dysosma difformis-an ethnomedicinal plant species were isolated to explore novel sources of podophyllotoxin. Fifty-three endophytic fungi were isolated and identified by morphological observation and ITS-based rDNA sequencing, assigning them to 27 genera in 3 divisions. Fusarium was found the most prevalent genus with a colonization frequency of 11.11%, followed by Trametes (9.26%) and Penicillium (7.41%). Phylogenetic trees were constructed for the endophytic fungi community in two collection sites, Ha Giang and Lai Chau, revealing the adaptation of the species to the specific tissues and habitats. Cytotoxic activity of endophytic fungal extracts was investigated on cancer cell lines such as SK-LU-1, HL-60, and HepG2, demonstrating strong anti-cancer activity of six isolates belonging to Penicillium, Trametes, Purpureocillium, Aspergillus, and Ganoderma with IC50 value of lower than 10 ㎍/mL. The presence of podophyllotoxin was indicated in Penicillium, Trametes, Aspergillus and for the first time in Purpureocillium and Ganoderma via high-performance liquid chromatography, which implied them as a potential source of this anticancer compound.

Bulb of Lilium longiflorum Thunb Extract Fermented with Lactobacillus acidophilus Reduces Inflammation in a Chronic Obstructive Pulmonary Disease Model

  • Ji-Eun Eom;Gun-Dong Kim;Young In Kim;Kyung min Lim;Ju Hye Song;Yiseul Kim;Hyeon-Ji Song;Dong-Uk Shin;Eun Yeong Lim;Ha-Jung Kim;Sung Hoon Kim;Deuk Sik Lee;So-Young Lee;Hee Soon Shin
    • Journal of Microbiology and Biotechnology
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    • v.33 no.5
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    • pp.634-643
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    • 2023
  • Chronic obstructive pulmonary disease (COPD), one of the leading causes of death worldwide, is caused by repeated exposure to harmful matter, such as cigarette smoke. Although Lilium longiflorum Thunb (LLT) has anti-inflammatory effects, there is no report on the fermented LLT bulb extract regulating lung inflammation in COPD. Thus, we investigated the protective effect of LLT bulb extract fermented with Lactobacillus acidophilus 803 in COPD mouse models induced by cigarette smoke extract (CSE) and porcine pancreas elastase (PPE). Oral administration of the fermented product (LS803) suppressed the production of inflammatory mediators and the infiltration of immune cells involving neutrophils and macrophages, resulting in protective effects against lung damage. In addition, LS803 inhibited CSE- and LPS-induced IL-6 and IL-8 production in airway epithelial H292 cells as well as suppressed PMA-induced formation of neutrophil extracellular traps in HL-60 cells. In particular, LS803 significantly repressed the elevated IL-6 and MIP-2 production after CSE and LPS stimulation by suppressing the activity of the nuclear factor kappa-light-chain-enhancer of activated B (NFκB) in mouse peritoneal macrophages. Therefore, our results suggest that the fermented product LS803 is effective in preventing and alleviating lung inflammation.