• Title/Summary/Keyword: human cell lines

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shRNA Mediated RHOXF1 Silencing Influences Expression of BCL2 but not CASP8 in MCF-7 and MDA-MB-231 Cell Lines

  • Ghafouri-Fard, Soudeh;Abdollahi, Davood Zare;Omrani, Mirdavood;Azizi, Faezeh
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.11
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    • pp.5865-5869
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    • 2012
  • RHOXF1 has been shown to be expressed in embryonic stem cells, adult germline stem cells and some cancer lines. It has been proposed as a candidate gene to encode transcription factors regulating downstream genes in the human testis with antiapoptotic effects. Its expression in cancer cell lines has implied a similar role in the process of tumorigenesis. The human breast cancer cell lines MDA-MB-231 and MCF-7 were cultured in DMEM medium and transfected with a pGFP-V-RS plasmid bearing an RHOXF1 specific shRNA. Quantitative real-time RT-PCR was performed for RHOXF1, CASP8, BCL2 and HPRT genes. Decreased RHOXF1 expression was confirmed in cells after transfection. shRNA knock down of RHOXF1 resulted in significantly decreased BCL2 expression in both cell lines but no change in CASP8 expression. shRNA targeting RHOXF1 was shown to specifically mediate RHOXF1 gene silencing, so RHOXF1 can mediate transcriptional activation of the BCL2 in cancers and may render tumor cells resistant to apoptotic cell death induced by anticancer therapy. shRNA mediated knock down of RHOXF1 can be effective in induction of apoptotic pathway in cancer cells via BCL2 downregulation, so it can have potential therapeutic utility for human breast cancer.

Cytotoxic Effect of the Distilled Pine-Needle Extracts on Several Cancer Cell Lines in vitro (솔잎 수액 증류액의 암세포주에 대한 in vitro 세포독성)

  • Chung, Young-Jin;Bae, Myung-Won;Chung, Myoung-Il;Lee, Ji-Seon;Chung, Kyeong-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.4
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    • pp.691-695
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    • 2002
  • This study was performed to examine the cytotoxic effects of the distilled pine-needle extracts against several cancer cell lines. First, cell lines including mice leukemic cancer cell line (L1210), sarcoma 180 and human monocyte-like cancer cells (U937) were tested using XTT methods in uitro. Pine-needle extracts were prepared by pressing the pine needles and distilling it at below 98$^{\circ}C$ and then added to the growth medium in a final dilution of 10, 20, and 40 times. Growth of three kinds of cancer cells was significantly inhibited by more than 50% with the addition of the extracts. Fifty six to seventy six % of inhibition was shown with the 40 times dilution of the extracts. Greater inhibition was achieved with the 20 times dilution (81~90%) and the 10 times dilution (77~89%) of the extracts. Next, other human cancer cell lines including 3 kinds of breast cancer cell lines (T47D, MDA-MB-231 and MW7A) and one hepatoma cell line (SNU-354) were tested with the 20 times dilution of the extract. T47D and MDA-MB-231 cell lines showed lower inhibition (12%) with the addition of the extract. However, MH7A and SNU-354 cell lines showed 64% and 72% inhibition with the extract, respectively. These results suggest that the distilled pine-needle extracts have strong cytotoxic effect on certain cancer cell lines and the intensity of the effect may vary depending on the process of the pine needle.

Cytokine Induction of Intercellular Adhesion Molecule-1(ICAM-1) Expression on Human Glioblastoma Cell Line, U-251 MG, U-373 MG (교모세포종 U-251MG, U-373MG세포주의 Cytokines처리에 의한 세포내 ICAM-1 발현)

  • Lee, Jong-Won;Kwon, Jung-Taek;Min, Byung-Kook;Park, Seung-Won;Kim, Young-Baeg;Hwang, Sung-Nam;Suk, Jong-Sik;Choi, Duck-Young
    • Journal of Korean Neurosurgical Society
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    • v.29 no.4
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    • pp.477-484
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    • 2000
  • Objective : Despite advances in the understanding of tumor biology and the tumor immunology, there has been no effective treatment. The Intercellular adhesion molecule-1(ICAM-1) has been shown to be important in interaction involving cells of the immune system and to be upregulated in a number of cell culture systems by cytokines, including immune interferon($IFN-{\gamma}$) and tumor necrosis $factor-{\alpha}$($TNF-{\alpha}$). ICAM-1 has been identified as one of the ligands for lymphocyte function-associated antigen-1(LFA-1). The effectiveness of various cytokines to ICAM-1 induction on cultured human glioblastoma cell lines and potential efficacy of immunotherapy were studied. Method : Human glioblastoma cell lines, U-251 MG, U-373 MG were trypsinized and suspended at $1{\times}10^5cells/ml$ and grown on 8 well chamber slide, the cells were incubated in 0.3ml medium alone or medium containing $IFN-{\gamma}$(1000U/ml) or $TNF-{\alpha}$(250U/ml) or $IFN-{\gamma}$ plus $TNF-{\alpha}$ for 6, 12, 24, 48 and 72 hours. The coverslip were then removed and stained with a 1/30 dilution of anti-ICAM-1 antibody. Result : Surface antigen expression of ICAM-1 was increased by incubating glioblastoma cell lines with $IFN-{\gamma}$ and $TNF-{\alpha}$. Combined effect of $IFN-{\gamma}$ and $TNF-{\alpha}$ has induced more ICAM-1 expression on glioblastoma cell lines. Upregulation of ICAM-1 expression in an established glioblastoma cell line was of greater magnitude and more rapid following incubation with $IFN-{\gamma}$ plus $TNF-{\alpha}$. Surface antigen expression of ICAM-1 was increased for up to 48 hours after cytokine treatment on both cell lines(p<0.05). There was no difference on both cell lines(p>0.05). Conclusion : The results of the present study indicate that ICAM-1 expression in glioblastoma cell lines, U-251 MG and U-373 MG, are induced and enhanced after treatment with $IFN-{\gamma}$ and $TNF-{\alpha}$. Combined effect of $IFN-{\alpha}$ and $TNF-{\gamma}$ is stronger and more rapid than $IFN-{\gamma}$ or $TNF-{\alpha}$ alone.

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Effects of Continentalic from Aralia Continentalis on Growth Inhibition and Apoptosis Induction in Human Leukemia HL-60 Cells (독활 유래 Continentalic Acid가 인간 백혈병 HL-60 세포의 성장억제와 아포토시스 유도에 미치는 영향)

  • Kim, Sun-Young;Jeong, Seung-Il;Kim, Sung-Zoo;Shim, Jae-Suk;Jang, Seon-Il
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.6
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    • pp.1314-1319
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    • 2009
  • In previous study, we have shown that continentalic acid (CA) isolated from Aralia continentalis induced the growth inhibition and apoptosis in HepG2 cells. In this study, we examine the effects of CA from A. continentalis on growth inhibition and apoptosis induction in human leukemia HL-60 and mouse fibroblast NIH 3T3 cell lines. The results demonstrated that CA decreased cell growth of leukemia HL-60 cells but not human HaCaT keratinocytes, assessed with the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] and LDH (lactate dehydrogenase) assay. Flow cytometric analysis of mouse fibroblast cell lines exposed to CA showed that apoptotic cells increased in a time- and dose-dependent manner. Treatment with CA decreased the number of normal cells and increased the number of early apoptotic and late apoptotic cells in a dose-dependent manner. The induction of apoptosis in mouse cell lines by CA was mediated through the activation of caspase-3, Bak, and Bax and the down-regulation of Bcl-2. Our results suggest that CA efficiently induces apoptosis in human leukemia cells.

Enhancement of Anti-tumorigenic Polysaccharide Production, Adhesion, and Branch Formation of Bifidobacterium bifidum BGN4 by Phytic Acid

  • Ku, Seock-Mo;You, Hyun-Ju;Ji, Geun-Eog
    • Food Science and Biotechnology
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    • v.18 no.3
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    • pp.749-754
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    • 2009
  • The polysaccharide (BB-pol) extracted from Bifidobacterium bifidum BGN4 showed growth inhibitory effects on several colon cancer cell lines such as HT-29 and HCT-116. To increase the yield of polysaccharide, B. bifidum BGN4 was cultured in various culture media with different compositions. When B. bifidum BGN4 was cultured in modified MRS broth containing phytic acid, the cells showed increased branch formation and enlarged morphology. The content of total carbohydrate and the ability of adhesion to intestinal epithelial cells were also increased by phytic acid. The polysaccharide obtained from the cells grown in the presence of phytic acid inhibited the proliferation of cancer cell lines such as HT-29 and MCF-7 cells but not normal colon cell line, FHC. Taken together, Bifidobacterium grown in the presence of phytic acid may confer enhanced beneficial function for the host.

The Combined Effect of Moschus and Anti-tumor drug Mitomycin C (사향과 항암제 Mitomycin C의 병용효과)

  • Eun Jae Soon;Kim Dae Keun;Song Jung Mo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.6
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    • pp.1404-1408
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    • 2003
  • The combined effects of water-soluble fraction of Moschus (ME) and anti-tumor drug mitomycin C on the proliferation of human tumor cell-lines were estimated by MTT colorimetric assay. ME inhibited the proliferation of Hep G2, A540, HeLa, KHOS-NP and Balb/c 3T3 cells. Also, ME increased the cytotoxicity of mitomycin C on Hep G2, A549 and HeLa cells. In addition, ME enhanced the cell viability of murine splenocytes and human lymphocytes at the concentration of 100㎍/㎖. These results indicate that ME inhibits the proliferation of human tumor cells and increases the cytotoxicity of mitomycin C without cytoxicity on immune cells.

Effective Chemopreventive Activity of Genistein against Human Breast Cancer Cells

  • Shon, Yun-Hee;Park, Sun-Dong;Nam, Kyung-Soo
    • BMB Reports
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    • v.39 no.4
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    • pp.448-451
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    • 2006
  • Chemopreventive and cytotoxic effect of genistein against human breast cancer cell lines was investigated. Genistein inhibited cell proliferation in estrogen receptor-positive (MCF-7) and estrogen receptor-negative (MDA-MB-231) human breast carcinoma cell lines. Cytochrome P450 (CYP) 1A1-mediated ethoxyresorufin O-deethylase (EROD) activity was inhibited by genistein in a concentrationdependent manner. Genistein significantly inhibited 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced cyclooxy-genase-2 activity and protein expression at the concentrations of 10 (p < 0.05), 25 (p < 0.05) and 50 mM (p < 0.01). In addition, ornithine decarboxylase (ODC) activity was reduced to 53.8 % of the control after 6 h treatment with 50 mM genistein in MCF-7 breast cancer cells. These results suggest that genistein could be of therapeutic value in preventing human breast cancer.

Effect of ${\alpha}$-Glycosidase Inhibitor in Multidrug Resistant Cell Lines

  • Paek, Nam-Soo;Namgung, Jun;Lee, Jung-Joon;Choi, Yong-Jin;Kim, Tae-Han;Kim, Kee-Won
    • BMB Reports
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    • v.31 no.3
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    • pp.269-273
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    • 1998
  • The objective of this study was to evaluate the reversal of multi drug resistance of human cell lines by specific inhibitors of ${\alpha}-glycosidase$ and mannosidases that had been reported to be involved in N-linked oligosaccharide processing of glycoproteins. N-methyldeoxynojirimycin, I-deoxynojirimycin, and castanospermine, which were known to be potent inhibitors of both ${\alpha}-glycosidase$ I and II, showed no activity against the multidrug resistant phenotype of the cell lines of SNU1DOX, KB-V1, and MCF-7/ADR. In contrast, I-deoxymannojirimycin, an inhibitor of mannosidase I, resulted in a slight reversal for the vinblastine resistance of the KB-V1 cell line, but did not show any activity toward the other cell lines. Parallel experiments with tunicamycin, an inhibitor of N-linked glycosylation, also resulted in no significant changes in multidrug resistant (MDR) phenotype of the cell lines tested in this work. These observations suggest that the unglycosylation of P-glycoprotein associated with the inhibitor treatments might not be correlated with the reversal of multidrug resistance of the cell lines tested in this study.

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The Inhibitory Effect of Ferulic Acid and Related Phenolic Compounds against Cancer Cell Lines (Ferulic Acid와 관련 페놀화합물의 암세포주에 대한 독성억제효과)

  • Han, Du Seok;Chun, Joo Won;Jeon, Sung Woo;Ba다, Seung Hwa
    • YAKHAK HOEJI
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    • v.49 no.5
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    • pp.365-369
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    • 2005
  • The inhibitory effect of ferulic acid and related phenolic compounds on the growth of normal cell lines and can­cer cell line was evaluated by the MTT and XTT methods. Ferulic acid decreased the cell viability of human skin melanoma cells by the MTT method and the cell adhesion activity of human oral epithelioid carcinoma cells by the XTT method. These results suggest that ferulic acid has a potential anticancer activity.