• Title, Summary, Keyword: HL-60 cells

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Induction of Apoptosis in Human Monocytes by Human Cytomegalovirus is Related with Calcium Increase

  • Moon, Myung-Sook;Lee, Gyu-Cheol;Lee, Chan H.
    • Journal of Microbiology
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    • v.40 no.3
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    • pp.224-229
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    • 2002
  • The effect of human cytomegalovirus (HCMV) on three human monocyte cell lines at different stages of differentiation was investigated. While the viability of HL-60 cells or U-937 cells was not significantly affected by HCMV infection, the viability of THP-1 cells was reduced. Acridine orange/ethidiurn bromide staining revealed that the reduction of THP-1 cell viability was due to increased apoptotic death following HCMV infection. Apoptosis in HL-60 cells was not affected by HCMV infection, and induction of apoptosis of U-937 cells by HCMV was intermediate between HL-60 and THP-1 cells. Since HL-60 cells are the least differentiated and THP-1 cells are the most differentiated, the induction of apoptosis of human monocytes appears to be related to the degree of cell differentiation. Flow cytometric and confocal microscopic studies using fluorescent calcium indicator Fluo-3 suggested a significant increase in intracellular free calcium concentration ([Ca$\^$2+/]i) in THP-1 cells undergoing apoptosis by HCMV infection. Again [Ca$\^$2+/]i in HCMV-infected HL-60 cells was not critically altered, and that in HCMV-infected U-937 cells was intermediate between THP-1 cells and HL-60 cells. Calcium influx blockers such as verapamil and nifedipine partially reversed HCMV-induced apoptosis in THP-1 cells.

Influence of Rubiae Radix Extract on the Mechanism of Apoptosis in HL-60 Cells (천초근 추출물이 HL-60 세포주의 세포자멸사 기전에 미치는 영향)

  • Choi, Ho-Seung;Park, Jin-Mo;Ju, Sung-Min;Kim, Sung-Hoon;Kim, Dae-Keun;Kim, Won-Sin;Jeon, Byung-Hun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.3
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    • pp.548-555
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    • 2008
  • Rubiae radix belonging to the family Rubiaceae have been used in traditional medicine to blood stasis and hemostasis. In this study, we reported that methanol extract of Rubiae radix (RRME) induced apoptotic cell death through MAPKs activation in human promylocytic leukemia (HL-60) cells. The cytotoxic activity of activity of RRME in HL-60 cells was increased in a dose-dependent manner. RRME was cytotoxic to HL-60 cells, with IC50 of $8{\mu}g/mL$. Treatment of RRME to HL-60 cells showed apoptotic bodies, and the fragmentation of DNA, suggesting that these cells underwent apoptosis. Caspase-3 activity and PARP cleavage were time-dependently increased the expression of Bcl-2 and Bax. And ratio of Bax/Bcl-2 protein expression. Activation of p38 and JNK were increased 6 hr after RRME treatment in HL-60 cells, but activation of ERK was reduced 24 hr after treatment. Taken together, these results suggest that RRME induces apoptotic cell death through activation of p38 and JNK in HL-60 cells.

Induction of Apoptosis by Extracts of Trichosanthes kirilpwii var. japonica in HL-60 Leukemia Cells (노랑하늘타리 추출물의 HL-60 혈액종양세포 Apoptosis 유도 효과)

  • 김상철;박수영;현재희;이영기;박덕배;강사윤;유은숙;강희경
    • YAKHAK HOEJI
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    • v.47 no.5
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    • pp.319-324
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    • 2003
  • This study examined the inhibitory effect of extracts of Trichosanthes kirilpwii sorted according to the parts on the growth of HL-60 cells. The growth of HL-60 leukemia cells was markedly inhibited by the treatment of the 80% methanol extract of roots (10 $\mu\textrm{g}$/mι), stems (50$\mu\textrm{g}$/mι), pips (10$\mu\textrm{g}$/mι), and gourds (100 $\mu\textrm{g}$/mι), or the ethylacetate fraction of leaves (100 $\mu\textrm{g}$/mι). when the HL-60 cells were treated with the extracts of T. kirilpwii sorted according to the parts, DNA fragmentation and sub-G1 hypodiploid cells were observed. Moreover, T. kirilpwii extracts increased the level of the expression of the active form of caspase-3 and the activation of caspase-3 was demonstrated by the cleavage of poly(ADP-ribose) polymerase, a vital substrate of effector caspase. The results suggest that the inhibitory effect of extracts of T. kirilpwii sorted according to the parts on the growth of HL-60 cells seems to arise from the induction of apoptosis.

Cytotoxicity in HL-60 cells and human lymphocytes and effect of leukemia cell differentiation induced by Saussureae Radix extract (정상사람림프구와 HL-60 cell에서 목향의 세포독성과 백혈병세포 분화효과에 관한 연구)

  • Lee, Young-Joon;Kang, Su-Jin;Ku, Sae-Kwang
    • The Korea Journal of Herbology
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    • v.26 no.2
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    • pp.31-37
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    • 2011
  • Objectives : This study was focused to investigate the toxicity of Saussurea lappa (SL) extracts in HL-60 cells and human lymphocytes. We also examined the differentiation effect of SL against leukemia cells. Methods : For examining the toxicity of SL, cytokinesis-block micronucleus (CBMN) assay and single cell gel eletrophoresis (SCGE) assay were used in present study. The cell differentiation effect of SL was evaluated by nitroblue tetrazolium (NBT) reduction assay. Results : The inhibition of cell growth in HL-60 cells was observed in a dose-dependant manner after SL treatment for 24 h. According to SCGE assay, HL-60 cells treated with SL increased DNA damage at $10{\mu}g/m{\ell}$, while DNA damage was induced by 0.1, 1, $10{\mu}g/m{\ell}$ concentration of SL in human lymphocytes. Our results indicated that SL have no genotoxic effect in HL-60 cells and human lymphocytes. Additionally, the differentiation effect was induced in $1{\mu}g/m{\ell}$ SL-treated HL-60 cells. Conclusions : From above results it is suggested that SL could be beneficial for the preparation of the useful agent for treating leukemia.

Terpinen-4-ol Induces Autophagic and Apoptotic Cell Death in Human Leukemic HL-60 Cells

  • Banjerdpongchai, Ratana;Khaw-on, Patompong
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.12
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    • pp.7537-7542
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    • 2013
  • Background: Terpinen-4-ol, a monoterpene, is found as the main component of essential oil extracts from many plants. In this study apoptotic and autophagic types of cell death induced by terpinen-4-ol and associated mechanisms were investigated in human leukemic HL-60 cells. Materials and Methods: The cytotoxicity of human leukemic U937 and HL-60 cells was determined by MTT assay. Cytochrome c release, expression of Bax, Bcl-2, Bcl-xl and cleaved Bid were determined by Western blotting. Cell morphology was examined under a transmission electron microscope. LC3-I/II, ATG5 and Beclin-1 levels were detected by immunoblotting. Results: Terpinen-4-ol exhibited cytotoxicity to human leukemic HL-60 but not U937 cells. The apoptotic response to terpinen-4-ol in HL-60 cells was due to induction of cytochrome c release from mitochondria and cleavage of Bid protein after the stimulation of caspase-8. There was a slightly decrease of Bcl-xl protein level. The characteristic cell morphology of autophagic cell death was demonstrated with multiple autophagosomes in the cytoplasm. At the molecular level, the results from Western blot analysis showed that terpinen-4-ol significantly induced accumulation of LC3-I/II, ATG5 and Beclin-1, regulatory proteins required for autophagy in mammalian cells. Conclusions: Terpinen-4-ol induced-human leukemic HL-60 cell death was via both autophagy and apoptosis.

Extract of Listeria monocytogenes Induces the Apoptosis on the Human Promyelocytic Leukemia Cells, HL-60 Cells (Listeria monocytogenes에 의해 HL-60 cell의 세포고사 유도 효과 규명)

  • Yang, Eun-Ju;Kim, Dong-Hyun;Chang, Jeong-Hyun
    • The Journal of the Korea Contents Association
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    • v.12 no.2
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    • pp.339-348
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    • 2012
  • Acute promyelocytic leukemia (APL) is a cancer of the blood and bone marrow. Although all-trans retionic acid (ATRA) is the agents for ALP therapy, there are various side effects. For overcome this problem, we need the development of new therapeutic agents for APL. A number of bacteria produce various virulence factors with cytotoxic effects on human cancer cells. To understand the anti-cancer effect of Listeria monocytogenes on APL, we examined alteration of the cell viability, apoptosis and cell cycle arrest of the human promyelocytic leukemia cell line, HL-60 cells. The cell supernatant (LmSup) and the extract of L. monocytogenes (LmE) inhibited the cell viability and induced apoptosis of HL-60 cells. These cytotoxic effect of LmSup and LmE mediated by modulation of cell cycle and ROS production. These results indicate that released or included bacterial molecules from L. monocytogenes have a cytotoxicity in HL-60 cells. Therefore, LmSup and LmE may be used as the potential target for the treatment of cancer induced by HL-60 cells.

Growth Inhibitory Effects of Sesamolin from Sesame Seeds on Human Leukemia HL-60 Cells (참깨에서 분리된 세사몰린의 백혈병 세포주 HL-60 생장억제 효과)

  • Kim, Kwan-Su;Kang, Sam-Sik;Ryu, Su-Noh
    • Korean Journal of Pharmacognosy
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    • v.34 no.3
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    • pp.237-241
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    • 2003
  • This study was carried out to test the growth inhibitory effects of sesamolin obtained from sesame seeds. Sesamolin inhibited the growth of human leukemia HL-60 cells in cultures and the synthesis of macromolecules in dose- and time-dependent manners. Sesamolin in the $60{\sims}100\;{\mu}g/ml$ range was cytostatic. At concentrations greater than $200\;{\mu}g/ml$ sesamolin was cytocidal to HL-60 cells and at $60\;{\mu}g/ml$ inhibited the synthesis of DNA, RNA and protein in HL-60 cells by 35.1, 6.1, and 5.3%, whereas at $200\;{\mu}g/ml$ these inhibitions were 86.8%, 81.5% and 96.7%, respectively. The inhibitory effect of sesamolin on DNA synthesis was irreversible.

Investigation of Chemotactic Activities in Differentiated HL-60 Cells by a Time-lapse Videomicroscopic Assay

  • Jung, Yun-Jae;Woo, So-Youn;Ryu, Kyung-Ha;Jang, Myoung-Ho;Miyasaka, Masayuki;Seoh, Ju-Young
    • IMMUNE NETWORK
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    • v.6 no.2
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    • pp.76-85
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    • 2006
  • Background: Chemotaxis is one of the cardinal functions of leukocytes, which enables them to be recruited efficiently to the right place at the right time. Analyzing chemotactic activities is important not only for the study on leukocyte migration but also for many other applications including development of new drugs interfering with the chemotactic process. However, there are many technical limitations in the conventional in vitro chemotaxis assays. Here we applied a new optical assay to investigate chemotactic activities induced in differentiated HL-60 cells. Methods: HL-60 cells were stimulated with 0.8% dimethylformamide (DMF) for 4 days. The cells were analyzed for morphology, flow cytometry as well as chemotactic activities by a time-lapse videomicroscopic assay using a chemotactic microchamber bearing a fibronectin-coated cover slip and an etched silicon chip. Results: Videomicroscopic observation of the real cellular motions in a stable concentration gradient of chemokines demonstrated that HL-60 cells showed chemotaxis to inflammatory chemokines (CCL3, CCL5 and CXCL8) and also a homeostatic chemokine (CXCL12) after DFM-induced differentiation to granulocytic cells. The cells moved randomly at a speed of $6.99{\pm}1.24{\mu}m/min$ (n=100) in the absence of chemokine. Chemokine stimulation induced directional migration of differentiated HL-60 cells, while they still wandered very much and significantly increased the moving speeds. Conclusion: The locomotive patterns of DMF-stimulated HL-60 cells can be analyzed in detail throughout the course of chemotaxis by the use of a time-lapse videomicroscopic assay. DMF-stimulated HL-60 cells may provide a convenient in vitro model for chemotactic studies of neutrophils.

Apoptosis Induction of HL-60 Leukemia Cells by Extract of Crinum asiaticum (문주란 추출물의 HL-60 백혈병 세포 Apoptosis 유도 효과)

  • Hyun, Jae-Hee;Kim, El-Vi-Ra;Kang, Jung-Il;Kim, Sang-Cheol;Yoo, Eun-Sook;Kang, Hee-Kyoung
    • YAKHAK HOEJI
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    • v.52 no.1
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    • pp.1-6
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    • 2008
  • The present study investigated the antiproliferative effects of Crnum asiaticum var. japonicum against HL-60 human leukemia cells. The 80% MeOH extract or several solvent fractions from the C. asiaticum inhibited the growth of HL-60 cells, whereas the growth of HEL-299 cells, human embryonic lung fibroblast, was scarcely inhibited. When the HL-60 cells were treated with the $CHCl_3$ fraction, the BuOH fraction, the EtOAc fraction and the $H_2O$ fraction, DNA ladder, chromatin condensation and increase of sub-G1 hypodiploid cells were observed. Furthermore, the $CHCl_3$ fraction and the BuOH fraction reduced Bc1-2 mRNA level, whereas Bax mRNA level was increased. These results suggest that the inhibitory effect of C. asiaticum on the growth of the HL-60 cell might be mediated through the induction of apoptosis via the down-regulation of Bc1-2. Taken together, components of C. asiaticum might have a therapeutic potential for the treatment of human leukemia.

Effect of AC-264, a Novel Indole Derivative, on Apoptosis in HL-60 Cells

  • Lee, Kyeong;Kwon, Ok-Kyoung;Xia, Yan;Ahn, Kyung-Seop
    • Bulletin of the Korean Chemical Society
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    • v.31 no.12
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    • pp.3777-3781
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    • 2010
  • The anticancer effect and apoptotic mechanism of a novel indole derivative AC-264, a lead derived from a chemical library, were investigated in human promyelocytic leukemia HL-60 cells. HL-60 cells treated with AC-264 at various concentrations showed the morphological features of apoptosis, such as plasma membrane blebbing and cell shrinkage. AC-264 exhibited cytotoxic effect in various cancer cell lines with different degrees of potency. Especially, AC-264 was effective on increasing the population of apoptotic cells in HL-60 cells, as detected by the number of cells stained with Annexin V and PI. Furthermore, AC-264 activated caspase-3 enzyme activity and induced internucleosomal DNA fragmentation. These results indicated that AC-264 produces anti-cancer effect via apoptotic cell death by activating caspase-3 and inducing internucleosomal DNA fragmentation in HL-60 cells.