• Title/Summary/Keyword: B16 cell lines

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Role of Tumor Necrosis Factor-Producing Mesenchymal Stem Cells on Apoptosis of Chronic B-lymphocytic Tumor Cells Resistant to Fludarabine-based Chemotherapy

  • Valizadeh, Armita;Ahmadzadeh, Ahmad;Saki, Ghasem;Khodadadi, Ali;Teimoori, Ali
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8533-8539
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    • 2016
  • Background: B-cell chronic lymphocytic leukemia B (B-CLL), the most common type of leukemia, may be caused by apoptosis deficiency in the body. Adipose tissue-derived mesenchymal stem cells (AD-MSCs) as providers of pro-apoptotic molecules such as tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), can be considered as an effective anti-cancer therapy candidate. Therefore, in this study we assessed the role of tumor necrosis factor-producing mesenchymal stem cells oin apoptosis of B-CLL cells resistant to fludarabine-based chemotherapy. Materials and Methods: In this study, after isolation and culture of AD-MSCs, a lentiviral LeGO-iG2-TRAIL-GFP vector containing a gene producing the ligand pro-apoptotic with plasmid PsPAX2 and PMDG2 virus were transfected into cell-lines to generate T293HEK. Then, T293HEK cell supernatant containing the virus produced after 48 and 72 hours was collected, and these viruses were transduced to reprogram AD-MSCs. Apoptosis rates were separately studied in four groups: group 1, AD-MSCs-TRAIL; group 2, AD-MSCs-GFP; group 3, AD-MSCs; and group 4, CLL. Results: Observed apoptosis rates were: group 1, $42{\pm}1.04%$; group 2, $21{\pm}0.57%$; group 3, $19{\pm}2.6%$; and group 4, % $0.01{\pm}0.01$. The highest rate of apoptosis thus occurred ingroup 1 (transduced TRAIL encoding vector). In this group, the average medium-soluble TRAIL was 72.7pg/m and flow cytometry analysis showed a pro-apoptosis rate of $63{\pm}1.6%$, which was again higher than in other groups. Conclusions: In this study we have shown that tumor necrosis factor (TNF) secreted by AD-MSCs may play an effective role in inducing B-CLL cell apoptosis.

Development of a Reporter System Monitoring Regulated Intramembrane Proteolysis of the Transmembrane bZIP Transcription Factor ATF6α

  • Kim, Jin-Ik;Kaufman, Randal J.;Back, Sung Hoon;Moon, Ja-Young
    • Molecules and Cells
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    • v.42 no.11
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    • pp.783-793
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    • 2019
  • When endoplasmic reticulum (ER) functions are perturbed, the ER induces several signaling pathways called unfolded protein response to reestablish ER homeostasis through three ER transmembrane proteins: inositol-requiring enzyme 1 (IRE1), PKR-like ER kinase (PERK), and activating transcription factor 6 (ATF6). Although it is important to measure the activity of ATF6 that can indicate the status of the ER, no specific cell-based reporter assay is currently available. Here, we report a new cell-based method for monitoring ER stress based on the cleavage of $ATF6{\alpha}$ by sequential actions of proteases at the Golgi apparatus during ER stress. A new expressing vector was constructed by using fusion gene of GAL4 DNA binding domain (GAL4DBD) and activation domain derived from herpes simplex virus VP16 protein (VP16AD) followed by a human $ATF6{\alpha}$ N-terminal deletion variant. During ER stress, the GAL4DBD-VP16AD(GV)-$hATF6{\alpha}$ deletion variant was cleaved to liberate active transcription activator encompassing GV-$hATF6{\alpha}$ fragment which could translocate into the nucleus. The translocated GV-$hATF6{\alpha}$ fragment strongly induced the expression of firefly luciferase in HeLa Luciferase Reporter cell line containing a stably integrated 5X GAL4 site-luciferase gene. The established double stable reporter cell line HLR-GV-$hATF6{\alpha}$(333) represents an innovative tool to investigate regulated intramembrane proteolysis of $ATF6{\alpha}$. It can substitute active pATF6(N) binding motif-based reporter cell lines.

The Inhibitory Effects of Cheonsagunja-tang and Leech on the IL-6, IL-16, GM-CSF mRNA Level by Human Epithelial Cells (천사군자탕(喘四君子湯)과 수질(水蛭)(마황초(麻黃炒))가 인간 기관지상피세포의 IL-6, IL-16, GM-CSF 발현에 미치는 영향)

  • Jung, Hee-Jae;Jung, Sung-Ki;Rhee, Hyung-Koo;Han, Dong-Ha
    • The Journal of Internal Korean Medicine
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    • v.22 no.4
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    • pp.601-611
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    • 2001
  • Objectives : We aimed to identify the dose-dependent inhibitory effects of Cheonsagunja-tang(喘四君子湯), leech(Hirudo medicinalis/水蛭) roasted with Ephedrae Herba(麻黃) on the mRNA expression of IL-6, IL-16, GM-CSF involved in the asthma model. Methods : In the study BEAS-2B cell lines, human epithelial cells were used. These cells were stimulated with tumor necrosis factor(TNF)-${\alpha}$ for artificial inflammatory expression. ${\beta}$-actin messenger RNA(mRNA) was used by internal standard. After 24 hours of the Cheonsagunja-tang, leech-treatment, total cellular RNAs were collected treating RNA zol directly on the living cells. Then the transcriptional activities of IL-6, 16 and GM-CSF were measured by RT-PCR with electrophoresis. Results: In the Cheonsagunja-tang study, the mRNA expression of IL-6 showed 30% transcriptional inhibitory effect compared to the control group in the $100{\mu}l/ml$ category(p<0.005). In the IL-16, there was 26%, 31% and 31% transcriptional inhibitory effect compared to the control groups in the $4{\mu}l/ml$, $20{\mu}l/ml$ and $100{\mu}l/ml$ categories, respectively(p<0.05). In the GM-CSF, the experimental group had 56% transcriptional inhibitory effect compared to the control group in the $100{\mu}l/ml$ category(p<0.001). In other concentrations, there was no inhibitory effect. In the leech study, the mRNA expression of IL-6 showed 37% transcriptional inhibitory effect compared to the control group in the $100{\mu}l/ml$ category(p<0.001). In the IL-16, there was 63% and 67% transcriptional inhibitory effect compared to the control groups in the $20{\mu}/ml$ and $100{\mu}/ml$ categories, respectively(p<0.001). In the GM-CSF, there was 64% and 68% transcriptional inhibitory effect compared to the control groups in the $20{\mu}l/ml$ and $100{\mu}l/ml$ categories, respectively(p<0.001). In other concentrations, there was no inhibitory effect. Conclusions : This study shows that Cheonsagunja-tang and leech roasted with Ephedrae Herba have dose-dependent inhibitory effects on the mRNA expression of IL-6, IL-16 and GM-CSF in BEAS-2B cell lines, human epithelial cells.

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Chemopreventive Actions of Blond and Red-Fleshed Sweet Orange Juice on the Loucy Leukemia Cell Line

  • Dourado, Grace KZS;Stanilka, Joy M;Percival, Susan S;Cesar, Thais B
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.15
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    • pp.6491-6499
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    • 2015
  • Background: Red-fleshed sweet orange juice (ROJ) comes from a new variety of citrus cultivated in Brazil that contains high levels of ${\beta}$-carotene and lycopene, and similar amounts of hesperidin (HSP) and nutrients, equivalently to blond orange juice (BOJ). Such bioactive compounds are associated with chemopreventive actions in several cancer cell lines. The purpose of this study was to examine the cytotoxicity, cell cycle, apoptosis, and cytokine secretion after BOJ, ROJ, and HSP treatment of a novel T acute lymphoblastic leukemia cell line, Loucy. Materials and Methods: Loucy cells were incubated for 24-h with BOJ, ROJ, and HSP, and the viability was measured using trypan blue. Cell cycling and apoptosis were assessed by propidium iodide (PI) and annexin V-FITC/PI flow cytometry, respectively. Secretion of cytokines $IL-1{\alpha}$, $IL1-{\beta}$, IL-2, IL-4, IL-6, IL-10, IL-17A, $IFN{\gamma}$, $TNF{\alpha}$, $TGF{\beta}$, $MIP{\alpha}$, and $MIP{\beta}$ was determined by ELISA array. Results: BOJ and ROJ treatments promoted Loucy cell cytotoxicity. Additionally, BOJ induced cell cycle arrest in the G0/G1 phase, and decreased the cell accumulation in the G2/M. ROJ decreased only the G0/G1 fraction, while HSP did not change the cell cycle. BOJ led to apoptosis in a different fashion of ROJ, while the first treatment induced apoptosis by increase of late apoptosis and primary necrotic fractions, the second increased early and late apoptosis, and primary necrotic fraction compared to positive controls. HSP had no effect on apoptosis. IL-6 and IL-10 were abrogated by all treatments. Conclusions: Taking together, these results suggest potential chemopreventive effects of BOJ and ROJ on Loucy cells.

Study on 2-thioDMNQ-S160, A Derivative of Shikonin, Antitumor Constituent of Lithospermum Erythrorhizon (자초의 항암성분 수도체 2-thioDMNQ-S160에 관한 연구)

  • Song Gyu Yong;Lee Hyo Jung;Khil Jae-Ho;Kim Sung Hoon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.3
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    • pp.542-546
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    • 2002
  • The purpose of this study was to investigate the effect of 2-thioDMNQ-S160, shikonin analogue isolated from Uthospermum Erythrorhizon, on the antitumor activity. In the present study, 2-thioDMNQ-S160 exhibited a significant cytotoxicity against L1210, A549, U937, and 816-BL6 cell lines with IC/sub 50/s of 2.4ug/ml, 2,0ugjml. 4ug/ml and 20 ug/ml, respectively. 2-ThioDMNQ-S160 strongly inhibited adhesion of B16-BL6 cells to gelatin and matrigel coated matrices. Also, 2-ThioDMNQ-S160 exhibited anti-invasive effect of B16-BL6 cells. The T/C% was 123 % in 2-ThioDMNQ-S160 treated group in S-180 bearing ICA mice. These results suggested that 2-thioDMNQ-S160 might be a potent candidate of cancer drug.

Cytotoxicity of Lignans from Lindera erytherocarpa Makino

  • Lee, Sang-Myung;Baek, Seung-Hwa;Lee, Choong-Hwan;Lee, Hyang-Burm;Kho, Yung-Hee
    • Natural Product Sciences
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    • v.8 no.3
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    • pp.100-102
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    • 2002
  • Three lignans were isolated from a methanol extract of Lindera erytherocarpa Makino (Lauraceae) are evaluated in vitro cytotoxicity using three cancer cell line assay. The compounds were identified as methyllinderone (1), linderone (2), and kanakugiol (3) by spectroscopic methods. Amongst the compounds, methyllinderone (1) showed significant cytotoxicity against mouse melanoma (B16-FlO), human acetabulum fibrosarcoma (HT1080), and choronic myelogenous leukemia (K562) cancer cell lines with $ED_{50}$ values of 2.2, 2.5, 8.3 ${\mu}g/ml$, respectively.

Screening of Cytotoxicity of Hexane Extracts from Cornis fructus

  • Chun, Hyun-Ja;Choi, Won-Hyung;Lee, Jeong-Ho;Lee, Ji-Su;Yang, Hyun-Ok;Baek, Seung-Hwa
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.272.1-272.1
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    • 2003
  • Cornis fructus were extracted by successive extraction and then fractionated with hexane extract to get active fractions. This study was performed to determine the cytotoxic effect of hexane extract from Cornis fructus on NIH 3T3 fibroblasts and cancer cell lines using MTT assay. Hexane extract showed cytotoxic effect against A549, B16 melanoma and MDA-MB-231. Further fractionation with hexane extract were performed to obtain effective fraction, fraction 3 showed the cytotoxic effect against A549 and MDA-MB-231 cell line.

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The Effect of Aster glehni Fr. Schm. Extracts on Whitening and Anti-Wrinkle (섬쑥부쟁이(Aster glehni Fr. Schm.) 분획물의 미백 및 주름 개선 효과)

  • Kim, Han-Hyuk;Park, Gun-Hye;Park, Kang-Soo;Lee, Jin-Young;Kim, Tae-Hoon;An, Bong-Jeun
    • Journal of Life Science
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    • v.20 no.7
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    • pp.1034-1040
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    • 2010
  • Plants and their extracts can be utilized as inexpensive and rich resources of active constituents in the cosmetic field, as well as the food, pharmaceutical and medicinal fields. Until now, Aster glehni Fr. Schm. had no known active effect, except on anti-oxidation, that was found during investigations for application in the cosmetic field. In this study, we examined the inhibition of enzymatic reactions to protein levels in inclusive B16F10 melanoma cell lines. Significant inhibition of enzymatic reactions was observed in the EtOAc extract, which advanced to tyrosinase protein and TRP-1 in the B16F10 melanoma cell line. These results indicated that the effect of EtOAc extract inhibited expressions of tyrosinase protein and TRP-1 in the B16F10 melanoma cell line by 30.5% and 41.5% at 100ug/ml respectively. On the other hand, antimicrobial activity was evaluated to the four fractions in normal flora of the skin. Hexane extract was only exhibited in the higher clear zone in all strains. In conclusion, any cosmeceutical effects of Aster glehni Fr. Schm. may have a potential meaning, as well as possible value for further studies regarding the effects of chemical constituents of Aster glehni Fr. Schm.

Anti-oxidant, anti-inflammatory and Whitening effect of Benincasa hispida seed extract (동과자 추출물의 항산화, 항염 및 미백 효능)

  • Park, Gyu-Ri;Lee, Ji-An
    • Journal of Convergence for Information Technology
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    • v.10 no.7
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    • pp.249-256
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    • 2020
  • To investigate the natural cosmetic ingredients of Benincasa hispida seed extract on skin care, we measured anti-oxidant and anti-inflammatory, and whitening effect. DPPH free radical scavenging activity was increased in a dose-dependent manner. The total phenolic content was higher in methanol extract (22.42±0.002 mgGAE/g) than water extract (9.77±0.002 mgGAE/g). MTT assay was demonstrated that the seed extract did not have a cytotoxic effect in RAW264.7 and B16F10 cell lines. We also examined to find out the inhibitory activity on NO production and secretion of TNF-α cytokine in LPS-induced RAW264.7 cells. In B16F10 melanoma cells, the seed extract significantly suppressed α-MSH induced melanin synthesis. Furthermore, westernblot analysis revealed that methanol extract dramatically downregulated the expression level of MITF, TRP-1 and TRP-2. Taken together, the B. hispida seed extract posses anti-oxidant, anti-inflammatory and skin whitening activities, which might provided its functional efficacy in cosmetic materials.

The Cytotoxic Activity of 13(E)-Labd-13-ene-8$\alpha$, 15-diol from Brachyglottis monroi

  • Lim, Jin-A;Kwag, Jung-Sook;Nigel, B.Perry;Baek, Seung-Hwa
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.179.2-179.2
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    • 2003
  • The cytotoxic activity of 13(E)-Labd-13-ene-8$\alpha$, 15-diol(1), isolated from the ethanol extract of Brachyglottis monroi was evaluated against tumor cell lines such as P388, SNU-C4 MDA-MB231, B 16 melanoma and A549 in vitro. By mean of spectral analysis particularly by the aid of various two dimensional NMR experiments, 1H-NMR and 13C-NMR signals of (1) was completely assigned, and thus the structure of (1) was established unambiguously. (omitted)

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