• Title/Summary/Keyword: HL60 cells

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Cytotoxicity Evaluation of Essential Oil and its Component from Zingiber officinale Roscoe

  • Lee, Yongkyu
    • Toxicological Research
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    • v.32 no.3
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    • pp.225-230
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    • 2016
  • Zingiber officinale Roscoe has been widely used as a folk medicine to treat various diseases, including cancer. This study aims to re-examine the therapeutic potential of co-administration of natural products and cancer chemotherapeutics. Candidate material for this project, ${\alpha}$-zingiberene, was extracted from Zingiber officinale Roscoe, and ${\alpha}$-zingiberene makes up $35.02{\pm}0.30%$ of its total essential oil. ${\alpha}$-Zingiberene showed low $IC_{50}$ values, $60.6{\pm}3.6$, $46.2{\pm}0.6$, $172.0{\pm}6.6$, $80.3{\pm}6.6$ (${\mu}g/mL$) in HeLa, SiHa, MCF-7 and HL-60 cells each. These values are a little bit higher than $IC_{50}$ values of general essential oil in those cells. The treatment of ${\alpha}$-zingiberene produced nucleosomal DNA fragmentation in SiHa cells, and the percentage of sub-diploid cells increased in a concentration-dependent manner in SiHa cells, hallmark features of apoptosis. Mitochondrial cytochrome c activation and an in vitro caspase-3 activity assay demonstrated that the activation of caspases accompanies the apoptotic effect of ${\alpha}$-zingiberene, which mediates cell death. These results suggest that the apoptotic effect of ${\alpha}$-zingiberene on SiHa cells may converge caspase-3 activation through the release of mitochondrial cytochrome c into cytoplasm. It is considered that anti-proliferative effect of ${\alpha}$-zingiberene is a result of apoptotic effects, and ${\alpha}$-zingiberene is worth furthermore study to develop it as cancer chemotherapeutics.

A Comparison of Functional Fragrant Components of Cymbidium (Oriental Orchid) Species (기능성 지표물질 확인을 위한 동양란 심비디움(Cymbidium) 향기 성분 비교 분석)

  • Kim, Sung Min;Jang, Eu Jean;Hong, Jong Won;Song, Sung Ho;Pak, Chun Ho
    • Horticultural Science & Technology
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    • v.34 no.2
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    • pp.331-341
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    • 2016
  • We analyzed the functional fragrant components of three species of Cymbidium oriental orchids using gas chromatography-mass spectrometry (GC/MS). For the comparative analysis, C. goeringii 'Minchunran', 'Jugeumhwa', C. forrestii 'Chwigae', 'Songmae', 'Yongja', and C. faberi 'Choemae', 'Namyangmae', 'Hwaja' were investigated. Major fragrant components detected by GC/MS were selected on the basis of more than 3% value according to the analysis of peak area (%). We found that ${\alpha}$-bergamotene, which has a cytotoxic effect on breast cancer, cervical cancer, and glioblastoma, and nerolidol, which induces apoptosis of human hepatoma cells (HepG2), inhibits the growth of Streptococcus mutans and babesiosis, and has antibacterial properties, are common substances produced by C. goeringii L. Nerolidol and ${\beta}$-bisabolene, which is cytotoxic and suppresses the growth of malignant melanoma cells (B16-F10), HepG2, and leukemia cells (HL-60, K562), are major substances in C. forrestii R. Furthermore, ${\alpha}$-pinene, which inhibits the growth of gliobastoma cells (SF-767) and inhibits the anti-inflammatory action of hepatoma cells (BEL-7402); 1,8-cineole, which is a potential therapeutic agent for the treatment of gastric ulcers; and 1,3,7-octatriene, which functions as a pheromone, are the most common substances in C. faveri R. Thus, substances identified as major fragrant components in oriental orchid species have multiple beneficial applications in human health. This research forms the basis for further studies of the roles of major fragrant components in oriental orchids.

Effects of Oxidative Stress on Apoptosis and Antioxidant Enzyme Levels

  • Kim, Choonmi;Lee, Ji-Young
    • Biomolecules & Therapeutics
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    • v.4 no.4
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    • pp.402-407
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    • 1996
  • Effects of oxidative stress on the induction of apoptosis and the activity of antioxidant enzymes were investigated in HL-60 cells using $H_2O$$_2$and cisplatin which generate oxygen species in the cell. Various concentrations of oxidants were treated to cells and at different incubation time, cells were harvested for assays. Cell viability, morphology by propidium iodide staining and DNA fragmentation by agarose gel electrophoresis were observed to determine whether they induce apoptosis. The activity of antioxidant enzymes such as superoxide dismutase and catalase was also measured to evaluate the cellular response to the oxidative damage. The results are as follows: $H_2O$$_2$ induced apoptosis at 10 $\mu$M after 6h incubation, while it took 12h for cisplatin. Both oxidants induced the superoxide dismutase activity at a tolerable low concentration. However, at a concentration which causes apoptotic cell death, the enzyme level was dropped markedly at first and then recovered to the normal level after which it declined again, probably due to cell death. On the other hand, changes in the activity of catalase were not significant at most concentrations except the statistically significant decrease at 24h after 10 $\mu$M-$H_2O$$_2$treatment. In this study, $H_2O$$_2$- and cisplatintreated cells showed similar results in apoptotic response and enzyme activities, suggesting that anticancer activity of cisplatin may be related, at least in part, to the production of oxygen free radicals.

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Water Extracts of Cultured Mountain Ginseng Stimulate Immune Cells and Inhibit Cancer Cell Proliferation

  • Oh, Chan-Ho;Kang, Pil-Sung;Kim, Jae-Whune;Kwon, Jin;Oh, Suk-Heung
    • Food Science and Biotechnology
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    • v.15 no.3
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    • pp.369-373
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    • 2006
  • Water extracts obtained from cultured mountain ginseng (CMG) were evaluated for their ability to stimulate immune cells and inhibit cancer cell proliferation. The lymphocyte subpopulation in mouse splenocytes in vivo was significantly increased by the administration of the CMG extract (27.4 mg/mouse). Interleukin-2 and ${\gamma}$-interferon in the mice serum increased up to 30% in CMG extract-treated mice. At a concentration of 1.37 mg/mL, nitric oxide increased up to 400% in the macrophage cell line treated with CMG extract. The CMG extract significantly retarded the proliferation of human acute promyelocytic (HL60), human histiocytic (U937), and mouse lymphocytic (L1210) leukemia cell lines in vitro at concentrations over 2.74-13.7 mg/mL. In addition, CMG extract treatments (1.37 mg/mL and 2.74 mg/mL) lead to the increased expression of the p53 gene and protein in cultured U937 leukemia cell lines. These results indicate that water extracts of CMG are capable of both immune cell stimulation and cancer cell growth inhibition.

Cloning of a Glutathione S-Transferase Decreasing During Differentiation of HL60 Cell Line (HL6O 세포주의 분화 시 감소 특성을 보이는 Glutathione S-Transferase의 클로닝)

  • Kim Jae Chul;Park In Kyu;Lee Kyu Bo;Sohn Sang Kyun;Kim Moo Kyu;Kim Jung Chul
    • Radiation Oncology Journal
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    • v.17 no.2
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    • pp.151-157
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    • 1999
  • Purpose : By sequencing the Erpressed Sequence Tags of human 걸ermal papilla CDNA library, we identified a clone named K872 of which the expression decreased during differentiation of HL6O cell line. Materials and Methods : K872 plasmid DNA was isolated according to QIA plasmid extraction kit (Qiagen GmbH, Germany). The nucleotide sequencing was performed by Sanger's method with K872 plasmid DNA. The most updated GenBank EMBL necleic acid banks were searched through the internet by using BLAST (Basic Local Alignment Search Tools) program. Nothern bots were performed using RNA isolated from various human tissues and cancer cell lines. The gene expression of the fusion protein was achieved by His-Patch Thiofusicn expression system and the protein product was identified on SDS-PAGE. Results : K872 clone is 1006 nucleotides long, and has a coding region of 675 nucleotides and a 3' non-coding region of 280 nucleotides. The presumed open reading frame starting at the 5' terminus of K872 encodes 226 amino acids, including the initiation methionine residue. The amino acid sequence deduced from the open reading frame of K872 shares $70\%$, identity with that of rat glutathione 5-transferase kappa 1 (rGSTKl). The transcripts were expressed in a variety of human tissues and cancer cells. The levels of transcript were relatively high in those tissues such as heart, skeletal muscle, and peripheral blood leukocyte. It is noteworthy that K872 was found to be abundantly expressed in coloreetal cancer and melanoma cell lines. Conclusion : Homology search result suggests that K872 clone is the human homolog of the rGSTK1 which is known to be involved in the resistance of cytotoxic therapy. We propose that meticulous functional analysis should be followed to confirm that.

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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.

Antibiotic Spectrum and Mechanism of Centipedin (Centipede Scolopendra subspinipes multilans L. KOCH로부터 정제된 항균 물질 Centipedin의 항균 Spectrum 및 작용 Mechanism 연구)

  • Kim, Ki-Tae;Hong, Sa-Weon;Won, Ho-Shik;Kim, Hyo-Joon;Park, Kyung-Bae;Cho, Key-Seung
    • Korean Journal of Microbiology
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    • v.34 no.1_2
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    • pp.31-36
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    • 1998
  • A broad spectrum of antibiotic action was studied with Centipedin purified from centipede Scolopendra subspinipes multilans L. Koch aganist gram-positive, gram-negative bacteria and fungi. The minimal inhibitory concentrations(MICs) were determined in liquid medium. The significant antibiotic activity was obtained aganist gram-negative Klebsiella pneumoniae ATCC 8308 responsible for causing infection at lung and intestine. The MIC value against Klebsiella pneumoniae ATCC 8308 was $2{\mu}g/ml$, and this Centipedin was active against Proteus vulgaris NRRL B-123. In addition, it has been shown that Centipedin blocks procaryotic RNA transcription and a little of DNA replication system in vitro. Centipedin did not exhibit any significant cytotoxicity against animal cells such as human blood leukemia (HL-60) and mouse B lymphocyte myeloma cell.

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Chemical Constituents from the Leaf and Twig of Acer okamotoanum Nakai and their Cytotoxicity

  • Jin, Wen-Yi;Min, Byung-Sun;Youn, Ui-Jung;Hung, Tran-Manh;Song, Kyung-Sik;Seong, Yeon-Hee;Bae, Ki-Hwan
    • Korean Journal of Medicinal Crop Science
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    • v.14 no.2
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    • pp.77-81
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    • 2006
  • As a result of cytotoxic compounds against cancer cell lines from natural sources, senven compounds were isolated from the leaf and twig of Acer okamotoanum Nakai. The compounds (1-7) were identified as ethyl gallate (1), methyl gallate (2), gallic acid (3), trans $resveratrol-3-O-{\beta}-D-glucopyranoside$ (4), acertannin (5), nikoenoside (6), and fraxin (7) by physicochemical and spectroscopic data (including mp, UV, IR, MS, $^1H-NMR,\;^{13}C-NMR$, DEPT, and HMBC) in comparison with those of published papers. All the compounds were tested for their cytotoxic activity against L1210, HL-60, K562, and B16F10 cancer cell lines in vitro by MTT assay method. Compounds 1-3 and 5 showed cytotoxic activity against all tested cell lines with $IC_{50}$ values ranged from 12.5 to $72.2\;{\mu}M$. Of the compounds, methyl gallate (2) exhibited the most potent cytotoxic activity against L1210, HL-60, K562, and B16F10 tumor cells with $IC_{50}$ values of 12.5, 48.3, 22.8, and $22.8\;{\mu}M$, respectively. Other compounds did not show any cytotoxic activity against four cancer cell lines.

Bioactive Constituents from the n-Butanolic Fraction of Aruncus dioicus var. kamtschaticus

  • Vo, Quoc Hung;Nguyen, Phi Hung;Zhao, Bing Tian;Thi, Yen Nguyen;Nguyen, Duc Hung;Kim, Won Il;Seo, U Min;Min, Byung Sun;Woo, Mi Hee
    • Natural Product Sciences
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    • v.20 no.4
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    • pp.274-280
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    • 2014
  • Six compounds were isolated from the n-BuOH fraction of the aerial parts of Aruncus dioicus var. kamtschaticus including: sambunigrin (1), prunasin (2), aruncide A (3), aruncide C (4), 1-O-caffeoyl-${\beta}$-D-glucopyranose (5), and caffeic acid (6). Their structures were confirmed by comparing the spectral data with those reported in the literature. The isolated compounds (1 - 6) were then examined for their cytotoxic effects towards MCF-7, HL-60, and HeLa cancer cell lines, as well as their DPPH radical scavenging activity. The results indicated that compound 4 possessed the strongest inhibitory effect toward HeLa cell line with $IC_{50}$ value of $5.38{\pm}0.92{\mu}M$. Compound 3 possessed selective cytotoxic activity on HL-60 cells with $IC_{50}$ value of $6.27{\pm}0.17{\mu}M$, compound 5 was found as the best in inhibiting proliferation with $IC_{50}$ value of $2.25{\pm}0.09{\mu}M$, and the other compounds showed significant inhibition with $IC_{50}$ values ranging from 6.10 to $11.27{\mu}M$. Compound 5 also displayed the strongest cytotoxic effect toward MCF-7 cell line ($IC_{50}$ $4.32{\pm}0.15{\mu}M$). Both 5 and 6 demonstrated strong radical scavenging activity ($IC_{50}$ $6.87{\pm}0.03$ and $4.33{\pm}0.22{\mu}M$, respectively). Compounds 1 and 5 were isolated for the first time from this plant.

Anti-inflammatory and Anti-cancer Effects of Sterol-rich Fraction from Nannochloropsis oculata by using Saponification (해양미세조류(Nannochloropsis oculata)로부터 saponification을 통한 Sterols 분획물의 항염증 및 항암 효능 평가)

  • Kim, Junseong;Lakmal, H.H. Chaminda;Lee, Ji-Hyeok;Lee, WonWoo;Jeon, You-Jin
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.47 no.6
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    • pp.770-775
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    • 2014
  • The green microalga of the genus Nannochloropsis (class Eustigmatophyceae) is a leading candidate for biofuel production due to its ability to accumulate high oil content (28.7% of cellular ash-free dry weight). We investigated the anti-inflammatory and anticancer activities of sterol-rich fraction from nannochloropsis oculata n-hexane (NOH) extract after saponification of the microalga. Among the fractions with n-hexane, chloroform and ethyl acetate, the n-hexane fraction showed the highest anti-inflammatory activity in LPS-stimulated RAW 264.7 macrophage as well as anticancer activity against human leukemia HL-60 cells without the cytotoxity. And the sterol-rich fraction was obtained from the n-hexane fraction by open silica column under the gradient solvent condition with 100% hexane (1L), hexane : ethyl acetate (20 : 1, 10 : 1, 5 : 1, 1 : 1, v/v). Among the four fractions (NOH-1~4), especially NOH-1 contained the highest content of sterols. NOH1 showed the highest HL-60 (about 85%) and NO inhibitory activities at the concentration of $100{\mu}g/mL$. These results demonstrated that the sterol-rich fraction from N. oculata might be a useful candidate as anti-inflammatory and anticancer agents for anti-inflammatory and anticancer activity.