• Title/Summary/Keyword: Breast (MCF-7) cell line

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Quantitative Assessment of Xenoestrogenic Environmental Pollutants using E-SCREEN Assay (E-SCREEN Assay를 이용한 내분비계 장애물질의 정량적 평가)

  • 오승민;이상기;정규혁
    • YAKHAK HOEJI
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    • v.44 no.5
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    • pp.416-423
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    • 2000
  • There is a growing concern that a wide variety of chemicals released into the environment can disrupt the endocrine system of fish, wildlife and humans. Endocrine disrupting chemicals (EDCs) include pesticides such as DDT lindane and atrazine, the food packaging chemicals, phthalates and bisphenol A, alkylphenol ethoxylate detergents and the chemical industry by-products, dioxins. Xenoestrogens in the environment have been argued about health risk, because of estrogen mimetic chemicals are exposed only small amounts to human. A number of in vivo and in vitro assays are now in use to assess the activity of xenoestrogens in the environment. A human breast cancer cell line (MCF-7) was used to develop in vitro screening assay for the detection of xenoestrogenic environmental pollutants. The E-SCREEN (MCF7-BUS) assay is proposed as a reliable, easy and rapid-to-perform method. To optimize and validate this method before it can be used routinely, several phenol compounds and pesticides suspected to be estrogenic were tested using I-SCREEN assay. The results showed that this method is a valuable tool for screening potential estrogen-mimicking environmental pollutants and quantitative determination of estrogeniciy.

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BAG5 regulates PTEN stability in MCF-7 cell line

  • Zhang, Ying;Gao, Haiyan;Gao, Haidong
    • BMB Reports
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    • v.46 no.10
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    • pp.490-494
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    • 2013
  • The phosphatase and tensin homolog deleted on chromosome 10 (PTEN) is a tumor-suppressing lipid phosphatase that is frequently absent in breast tumors. Thus, the stability of PTEN is essential for tumor prevention and therapy. The ubiquitin-proteasome pathway has an important role in regulating the functions of PTEN. Specifically, carboxyl terminus Hsp70-interacting protein (CHIP), the E3 ubiquitin ligase of PTEN, can regulate PTEN levels. In this study, we report that BCL-2-associated athanogene 5 (BAG5), a known inhibitor of CHIP activity, reduces the degradation of PTEN and maintains its levels via an ubiquitylation-dependent pathway. BAG5 is identified as an antagonist of cell tumorigenicity.

Caveolin-1, Through its Ability to Negatively Regulate TLR4, is a Crucial Determinant of MAPK Activation in LPS-challenged Mammary Epithelial Cells

  • Wang, Xiao-Xi;Wu, Zheng;Huang, Hui-Fang;Han, Chao;Zou, Wei;Liu, Jing
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.4
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    • pp.2295-2299
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    • 2013
  • Background: To explore the role of caveolin-1(CAV-1) gene silencing on MAPK activation in lipopolysaccharide (LPS)-challenged human mammary epithelial cells. Methods: We established a MCF-10ACE of CAV-1 gene silencing from human mammary epithelial cell line MCF-10A by RNAi technology. DNA Microarray were used to detect the expression of inflammation-associated genes in MCF10ACE. Western blotting was used to examine the activation of MAPK in lipopolysaccharide(LPS)-challenged MCF-10A and MCF-10ACE. Moreover, immunofluorescence and Western bloting were performed to detect the co-localization of CAV-1 and toll-like receptor 4 (TLR4) in human mammary epithelial cells. Results: MCF-10ACE exhibited significant increases in inflammation-associated gene expression, especially IL-6 (~7-fold) and IL6R (~17-fold). In addition, LPS-induced p38 MAPK and JNK MAPK activation was significantly increased in MCF-10ACE. Furthermore, CAV-1 co-localized with TLR4 and appeared a negative correlation trend. Conclusion: CAV-1 gene silencing promotes MAPK activation via TLR4 signaling in human mammary epithelial cells response to LPS.

Bioassays of Polycyclic Aromatic Hydrocarbons in ZR-75-1 Human Breast Cancer Cells

  • Min, Kyung-N.;Sheen, Yhun-Y.
    • Environmental Mutagens and Carcinogens
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    • v.23 no.1
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    • pp.35-40
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    • 2003
  • Polycyclic aromatic hydrocarbons (PAH) are frequently detected in food, water, soil, and sediment and are widespread environmental pollutants formed by the incomplete combustion of fossil fuels, woods and other organic matter. PAHs are considered to be probable human carcinogens. The mechanism of action of PAHs has been studied extensively, however it is not clear how PAHs turn on CYP1A1 in human breast cancer. Our laboratory have been studied the effect of PAHs in the human breast cancer cell MCF7. In this study, we examined the ZR-75-1 human breast cancer cells as a new system to evaluate bioactivity of PAHs. ZR-75-1 human breast cancer cell line responses to estrogen and progesteron. We have been able to estbilish long term culture system of this cells then used for the study to observe the effect of PAHs. We demonstrate that PAHs induced the CYP1A1 promoter and 7-ethoxyresolufin O-deethylase (EROD) activity in a concentration-dependant manner. RT-PCR analysis indicated that PAHs significantly up-regulate the level of CYP1A1 mRNA. Some of PAHs showed stronger stimulatory effect on CYP1 gene expression than TCDD. Apparently, ZR-75-1 cells have Aryl hydrocarbon receptors, therefore it would be good experimental tool to study the cross-talk between PAHs and steroid actions.

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Effect of Soy Isoflavones on the Expression of $TGF-{\beta}1$ and Its Receptors in Cultured Human Breast Cancer Cell Lines

  • Kim Young-Hwa;Jin Kyong-Suk;Lee Yong-Woo
    • Biomedical Science Letters
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    • v.11 no.2
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    • pp.175-183
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    • 2005
  • The two major isoflavones in soy, genistein and daidzein, are well known to prevent hormone-dependent cancers by their anti estrogenic activity. The exact molecular mechanisms for the protective action are, however, not provided yet. It has been reported that genistein and daidzein have a potential anticancer activity through their antiproliferative effect in many hormone-dependent cancer cell lines. Transforming growth $factor-\beta1(TGF-\beta1)$ has also been found to have cell growth inhibitory effect, especially in mammary epithelial cells. This knowledge led to a hypothetical mechanism that the soy isoflavones-induced growth inhibitory effect can be derived from the regulation of $TGF-\beta1$ and $TGF-\beta$ receptors. In order to test this hypothesis, the effects of the soy isoflavones at various concentrations and periods on the expression of $TGF-\beta1$and $TGF-\beta$ receptors were investigated by using Northern blot analysis in human breast carcinoma epithelial cell lines, an estrogen receptor positive cell line (MCF-7) and an estrogen receptor negative cell line (MDA-MB-231). As a result, only genistein has shown a profound dose-dependent effect on $TGF-\beta1$ expression in the $ER^+$ cell line within the range of doses tested, and the expression levels are correspondent to their inhibitory activities of cell growth. Moreover, daidzein showed down-regulated $TGF-\beta1$ expression at a low dose, the cell growth proliferation was promoted at the same condition. Therefore, antiproliferative activity of the soy isoflavones can be mediated by $TGF-\beta1$ expression, and the effects are mainly, if not all, occurred by ER dependent pathway. The expression of $TGF-\beta$ receptors was induced at a lower dose than the one for $TGF-{\beta}1$ induction regardless of the presence of ER, and the expression patterns are similar to those of the cell growth inhibition. These results indicated that the regulation of $TGF-\beta$ receptor expression as well, prior to $TGF-\beta1$ expression, may be involved in the antiproliferative activity of soy isoflavones. Little or no expression of $TGF-\beta$ receptors was found in the MCF-7 and MDA-MB-231 cells, suggesting refractory properties of the cells to growth inhibitory effect of the $TGF-\beta$. The soy isoflavones can seemingly restore the sensitivity of growth inhibitory responses to $TGF-\beta1$ by re-inducing $TGF-\beta$ receptors expression. In conclusions, our findings presented in this study show that the antitumorigenic activity of the soy isoflavones could be mediated by not only $TGF-\beta1$induction but $TGF-\beta$ receptor restoration. Thus, soy isoflavones could be good model molecules to develop new nonsteroidal antiestrogenic chemopreventive agents, associated with, regulation of $TGF-\beta$ and its receptors.

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Antiproliferation effects of ethanol extract of garlic peels on human cancer cell lines (마늘껍질 70% 에탄올 추출물의 인간 암세포 증식억제 활성)

  • Son, Dae-Yeul
    • Food Science and Preservation
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    • v.24 no.2
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    • pp.289-293
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    • 2017
  • Ethanol extract of garlic peels (GPE) was investigated for its antiproliferative effects on human cancer cell lines. Human lung cancer cell line A549 treated with $500{\mu}g/mL$ GPE resulted in the growth inhibition of A549 by 90%. In stomach cancer cell AGS proliferation inhibition activity, GPE showed 45% and 71% inhibition of AGS growth at $1,000{\mu}g/mL$ and $2,000{\mu}g/mL$, respectively. GPE inhibited the growth of the breast cancer cells MCF-7 effectively at low concentration and showed 78% and 90% inhibitions of MCF-7 growth at $200{\mu}g/mL$ and $500{\mu}g/mL$, respectively. GPE showed very significant antiproliferation effect on liver cancer cell line Hep3B and inhibited Hep3B cell growth by 57% at $100{\mu}g/mL$, and the inhibition's rate increased up to 87% at $500{\mu}g/mL$. Antiproliferation effect of GPE on colorectal cancer cell HT-29 showed 15% reduction of HT-29 cell growth at $200{\mu}g/mL$ and the growth rate was reduced in a dose dependent manner up to $1,000{\mu}g/mL$. These results indicated that GPE had high antiproliferation effects on breast and liver cancer cell lines at low concentrations ($200{\mu}g/mL$), and by higher concentrations over $500{\mu}g/mL$, GPE inhibited the growth of A549 and HT-29. The results of our study suggested the potential use of garlic peels for use as an excellent antiproliferative substance for human cancer cells.

The Effect of Hwangryunhaedoktang on Proliferations of Various Human Cancer Cells (황련해독탕이 수종의 인간 암세포 증식에 미치는 영향)

  • Sung, Hyun Kyung;Min, Sang Yeon;Kim, Jang Hyun
    • The Journal of Pediatrics of Korean Medicine
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    • v.27 no.1
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    • pp.59-68
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    • 2013
  • Objectives The aim of this study is to investigate whether hwang-ryun-haedok-tang (HDT) affect proliferations of androgen-dependent LNCaP prostate cancer cells, androgen-independent PC-3, DU-145 prostate cancer cells, MCF-7 human breast cancer cells, A549, NCI-H292 human pulmonary cancer cells and K-562 human chronic myelogenous leukemia cells. Materials and Methods Effects of HDT on proliferations of each cancer cell line were investigated. 20,000 cells/well were plated in each well of 96-well culture plate. After 24 hrs, 0.01-10% of HDT in culture medium was added to cancer cells. The number of cells was counted by using SRB assay or direct cell counting method after 72 hours from drug treatment. Effect of baicalein or berebrine on proliferation was assessed according to the same method. Results (1) HDT inhibited proliferations of LNCaP, PC-3 and DU-145 prostate cancer cells. (2) HDT inhibited proliferation of MCF-7 breast cancer cells. (3) HDT also inhibited proliferations of A549, NCI-H292 pulmonary cancer cells and K-562 chronic myelogenous leukemia cells. (4) Baicalein and berberine also showed inhibitory effects on proliferations of prostate and breast cancer cells. Conclusion : HDT inhibited proliferations of human prostate, breast, pulmonary and blood cancer cells. These results suggest us the potential use of HDT as a chemopreventive or chemotherapeutic agent. Effect of HDT on human cancer should be further investigated using in vivo experimental models that can reflect pathophysiology of human cancer through another studies.

Translocation of Protein Kinase C Isozymes in the Breast Cancer Cell Line (유방 암세포에서 Protein Kinase C 동위효소의 전위)

  • Won Chul Choi;Joo Young Son;Seok Jin Seo
    • Journal of Life Science
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    • v.8 no.6
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    • pp.638-647
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    • 1998
  • Protein Kinase C (PKC) activators, phorbol 12-myristate 13-acetate (PMA), bryostatin, and dioctanoyl glycerol (DiC8), induce translocation of PKC isozymes from cytoplasm to plasma membrane or into nucleus. The activated PKC negatively modulates growth of human breast cancer cells. Antiproliferative effect and translocation of PKC were investigated in MCF-7 cells. The translocation of activated PKC isozymes by PMA, bryostatin and DiC8 was occurred at the various different regions in MCF-7 cell. PKC $\alpha$ and $\beta$ could be translocated to the nucleus or the nuclear mem-brane, and PKC $\delta$and $\varepsilon$ to cell membrane by PMA while DiC8 and bryostatin induced the translocation of PKC $\alpha$ and $\beta$ to the nucleus or plasma membrane, respectively. In the antiproliferative effect of PKC activators, PMA ($IC_{50}$/ values of 1.2$\pm$0.3nM) and DiC8 ($IC_{50}$/ values of 5.0$\pm$1.1$\mu$M) inhibited the cell growth. Bryostatin also inhibited the cell growth but to a much less degree than one obser-ved with PMA : 16% growth reduction by 100nM bryostatin. However, PMA treated with bryostatin induced gro-wth inhibition, but PMA with DiC8 at 10$\mu$M was not effective. These results suggest that each PKC isozyme is tran-slocated to various specific sites, and that especially, PKC $\alpha$ isozyme plays an important role in control of antiprolife-raive function of cell growth.

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In-vitro Anticancer and Antioxidant Activity of Gold Nanoparticles Conjugate with Tabernaemontana divaricata flower SMs Against MCF -7 Breast Cancer Cells

  • Preetam Raj, J.P;Purushothaman, M;Ameer, Khusro;Panicker, Shirly George
    • Korean Chemical Engineering Research
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    • v.54 no.1
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    • pp.75-80
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    • 2016
  • Biologically stabilized gold nanoparticles were synthesized from the flower aqueous extract of T. divaricata. The synthesized nanoparticles were characterized by UV-Vis spectrophotometer, Zeta sizer, FTIR and TEM analysis. T. divaricata reduced gold nanoparticles having particle size and potential of 106.532 nm and -10.2 mV, respectively, with a characteristic peak of 550 nm in UV-visible spectrophotometer. FTIR graph after comparison between the crude flower extract and gold nanoparticles showed three major shifts in the functional groups. The morphology and size of the gold nanoparticles were examined by HRTEM analysis, which showed that most of the nanoparticles were nearly spherical with size of 100 nm. The gold nanoparticles synthesized demonstrated potent anticancer activity against MCF-7 cell line. The findings conclude that the antioxidant molecule present in T. divaricata may be responsible for both reduction and capping of gold nanoparticles which possess potential applications in medicine and pharmaceutical fields.

Evaluation of antibacterial activity and cytotoxic effects of green AgNPs against Breast Cancer Cells (MCF 7)

  • Vizhi, Dhandapani Kayal;Supraja, Nookala;Devipriya, Anbumani;Tollamadugu, Naga Venkata Krishna Vara Prasad;Babujanarthanam, Ranganathan
    • Advances in nano research
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    • v.4 no.2
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    • pp.129-143
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    • 2016
  • The present work reports a facile, rapid and an eco-friendly method for the synthesis of silver nanoparticles using Luffa acutangula (L. acutangula) leaves extract and their antibacterial and cytotoxic effects. The synthesized silver nanoparticles (AgNPs) were characterized by UV-Visible spectroscopy (UV-Vis), Fourier transform infrared spectroscopy (FT-IR) and X-ray diffraction analysis (XRD). Additionally the topography, morphology and the elemental composition of the particles were determined by Scanning Electron Microscopy (SEM) and Energy dispersive spectrophotometric (EDS) technique and the measured particle sizes from SEM micrographs are in the range of 12.5 to 24.5nm. The in-vitro antimicrobial activity of the synthesized nanoparticles was high against gram positive Staphylococcus aureus and moderate against gram negative Escherichia coli and Pseudomonas aeruginosa strains. Further, the cytotoxic effects of synthesized AgNPs were evaluated against Human Breast Cancer (MCF 7) cell line.