• Title/Summary/Keyword: Cell-based assay

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Quinone Reductase Inductive Activity and Growth Inhibitory Effect against Hepatoma Cell of Oriental Melon Extract (참외 추출물의 Quinone Reductase 유도활성 및 간암세포 증식 억제효과)

  • Kim, Hye-Suk;Ku, Kang-Mo;Suh, Jun-Kyu;Kang, Young-Hwa
    • Journal of Bio-Environment Control
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    • v.18 no.4
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    • pp.448-453
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    • 2009
  • This study was performed to elucidate anticancer activities of various parts, such as peel, flesh, placenta, seed, stalk and stem leaf of oriental melon. Chemopreventive and anticancer effects of oriental melon extract were evaluated by detoxifying enzyme, quinone reductase (QR) inductive activity, cytotoxicity and growth inhibitory effect against hepatoma cell. Stalk and stem leaf extracts of oriental melon showed the increment of QR inductive activity with dose-dependent manner and induced quinone reductase 3.9, 1.5-fold at $200{\mu}g/mL$ respectively compared to control. The growth inhibitory effect of oriental melon extract against mouse hepatoma cell (Hepa1c1c7) was investigated by crystal violet (CV) assay. Stalk and stem leaf of oriental melon showed potent growth inhibitory effect. Based on these result, the growth inhibitory effects of stalk, stem leaf at various concentration were examined in detail by MTT assay using human hepatoma cancer cell (HepG2). All of two parts showed growth inhibitory effects and expecially stalk exhibited inhibitory effect of 60.3% at maximum concentration. The above results suggest that stalk of oriental melon has a possibility as a source of natural cancer chemopreventive materials.

The Effects of Anticarcinogenic Activity of Solanum tuberosum Peel Fractions (감자 껍질 분획성분의 항발암 효과)

  • 배송자
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.5
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    • pp.905-909
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    • 2002
  • This study was peformed to determine the anticarcinogenic activity of the Solanum tuberosum Peel (SP) on several microorganisms and human cancer cell lines. Among the various solvent fractions of SP, the ethylether Partition layer (SPMEE) showed the strongest antimicrobial activity, ethylacetate partition layer (SPMEA) and butanol partition layer (SPMB) resulted in good antimicrobial activity. We also determined the effect of SP extract and fractions on cytotoxicity, and chemopreventive effect on human cancer cells. The experiment was conducted to determine cytotoxicity of SP partition layers on HepG2, HeLa and MCF-7 cells by MTT assay. Among the various partition layers of SP, SPMEE and SPW were showed the strongest cytotoxic effects on all cancer cell lines. The quinone reductase induced activities of HepG2 cell, the butanol partition layer (SPMB) at a does of 40 $\mu\textrm{g}$/mL was 8.49 times more effective compared to the control value of 1.0. This value was significantly higher than that of previous results using the other materials. Therefore, based on these studies, SP may be developed into a potentially useful antimicrobial and anticarcinogenic agents.

Protective Effects of Hemerocallis Fulva Extracts on Amyloid $\beta$-Protein-Induced Death in Neuronal Cells (아밀로이드 베타 단백질에 의해 유도된 신경세포 독성에 대한 원추리의 억제 효과 탐색)

  • Kim Eun-Sook;Choi Soo-Jin;Ryu Beung-Ho;Choi Jin-Ho;Oh Myung-Sok;Park Woo-Jin;Choi Young-Whan;Paik Do-Hyeon;Ha Kwon-Chul;Kang Dae-Ook;Cho Yong-Kweon;Park Ki-Tae;Moon Ja-Young
    • The Journal of Korean Medicine
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    • v.27 no.2 s.66
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    • pp.122-133
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    • 2006
  • Objectives : The amyloid $\beta$-protein ($A\beta$) is the principal component of the senile plaques characteristic of Alzheimer's disease (AD) and elicits a toxic effect on neurons in vitro and in vivo. Many environmental factors including antioxidants and proteoglycans modify $A{\beta}toxicity$. In this study, we have investigated the protective effects of water- and organic solvent-extracts of Hemerocallis fulva root fractions pre-extracted with methanol on $A\beta$-induced oxidative cell death in cultured rat pheochromocytoma (PC12) cells. Methods : For this study, we used MTT reduction assay for detection of protective effects of water- and organic solvent-extracts of Hemerocallis fulva root fractions pre-extracted with methanol on $A{\beta}_{25-35}$-induced cytotoxicity to PC12 cells. We also used cell-based $\beta$-secretase assay system to investigate the inhibitory effect of water- and organic solvent-extracts of Hemerocallis fulva root on $\beta$-secretase activity. Results : We previously reported that methanol extracts of Hemerocallis fulva root strongly attenuated cytotoxicity induced by the three $A\beta$ fragments ($A{\beta}_{25-35},\;A{\beta}_{1-42}\;A{\beta}_{1-43}$) to both SK-N-MC and PC12 cells. In the present study, we found that butanol-, ethylacetate-, chloroform-, and water-extracts of Hemerocallis fulva root fractions pre-extracted with methanol had strong protective effects against $A{\beta}_{25-35}$-induced cytotoxicity to PC12 cells and inhibitory potency to $\beta$-secretase activity. Conclusion : These results suggest that butanol-, ethylacetate-, chloroform-, and water-extracts of Hemerocallis fulva root fractions pre-extracted with methanol may contain the protective component(s) against $A\beta$-induced cell death in PC12 cells as well as inhibitory component(s) to $\beta$-secretase activity.

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Cytoprotective Effect of Ethanol Extract from Maesil (Prunus mume Sieb. et Zucc.) on Alloxan-induced Oxidative Damage in Pancreatic-cell, HIT-T15 (Alloxan에 의한 HIT-T15 세포의 산화적 손상에 대한 매실(Prunus mume Sieb. et Zucc.) 주정추출물의 세포보호효과)

  • Kim, In-Hye;Kim, Jong-Bae;Cho, Kang-Jin;Kim, Jae-Hyun;Om, Ae-Son
    • Korean Journal of Plant Resources
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    • v.25 no.2
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    • pp.184-192
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    • 2012
  • The present study was designed to examine the potential antidiabetic and antioxidant effect of ethanol extract from $Prunus$ $mume$ fruit (PME) against alloxan-induced oxidative stress in pancreatic ${\beta}$-cells, HIT-T15. To evaluate the antidiabetic effect of PME, 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazoliu bromide (MTT) cell proliferation assay, lactate dehydrogenase (LDH) release assay, $NAD^+$/NADH ratio and insulin secretion were assessed. We also measured its antioxidant effect against alloxan-induced oxidative stress in the cells by assessing the levels of the antioxidant enzymes including superoxide dismutase (SOD), glutathione S-transferase (GST), glutathione reductase (GR) and glutathione peroxidase (GPx). The results of this analysis showed that alloxan significantly decreased cell viability, increased LDH leakage, and lowered $NAD^+$ /NADH ratio and insulin secretion in HIT-T15 cells. However, PME significantly increased the viability of alloxan-treated cells and lowered LDH leakage. The intracellular $NAD^+$ /NADH ratio and insulin secretion were also increased by 1.5~1.9-fold and 1.4-fold, respectively, after treatment with the PME. The HIT-T15 cells treated with alloxan showed significant decreases in the activities of antioxidant enzymes, while PME significantly elevated the levels of antioxidant enzymes. Based on these results, we suggest that PME could have a protective effect against the cytotoxicity and dysfunction of pancreatic ${\beta}$-cells in the presence of alloxan-induced oxidative stress.

The Significance of p53 Expression in Serum and Tissue from Patients with Lung Cancer (원발성 폐암환자의 혈청 및 조직에서의 p53단백 표현)

  • Chang, Jung-Hyun;Sung, Sun-Hee
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.2
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    • pp.333-340
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    • 1998
  • Background: Lung cancer is the leading cause of cancer over the world. P53 alteration is by far the most common genetic defect in lung cancer. The mutation of p53 protein involves the loss of inhibitory function of p53 related tumor suppressor gene and resultant oncogenesis. The analysis of p53 alterations consists of immunohistochemical stain, PCR based assay, or serologic ELISA (enzyme-linked immunosorbent assay). Methods : Serum levels of p53 mutant protein were measured in 69 cases of lung cancer (adenocarcinoma n=29, epidermoid n=16, small cell n=13, large cell n=1, undifferentiated n=1, undetermined n=9) and 42 controls of respiratory disorders using ELISA. Immunohistochemical stain in tissue was performed using monoclonal antibody of p53 in lung cancer subjects. Results: Both serum p53s in nonsmall cell cancer ($0.28{\pm}0.44ng/ml$) and in small cell cancer ($0.20{\pm}0.14ng/ml$) were not different from controls ($0.34{\pm}0.20ng/ml$). Also there was no significant difference in serum p53 according to tumor stages. P53 immunohistochemical stain showed 50% positivity overall in lung cancer. There were no close correlation between serologic level and positivity of immunohistochemical stain. Conclusion: The serologic determination of p53 mutant protein is thought to have no diagnostic role in lung cancer. Immunohistochemical stain in lung cancer specimen shows 50% positivity.

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LncRNA H19/miR-29b-3p/PGRN Axis Promoted Epithelial-Mesenchymal Transition of Colorectal Cancer Cells by Acting on Wnt Signaling

  • Ding, Dayong;Li, Changfeng;Zhao, Tiancheng;Li, Dandan;Yang, Lei;Zhang, Bin
    • Molecules and Cells
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    • v.41 no.5
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    • pp.423-435
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    • 2018
  • This investigation was aimed at working out the combined role of lncRNA H19, miR-29b and Wnt signaling in the development of colorectal cancer (CRC). In the aggregate, 185 CRC tissues and corresponding para-carcinoma tissues were gathered. The human CRC cell lines (i.e. HT29, HCT116, SW480 and SW620) and normal colorectal mucosa cell line (NCM460) were also purchased. Si-H19, si-NC, miR-29b-3p mimics, miR-29b-3p inhibitor, si-PGRN and negative control (NC) were, respectively, transfected into the CRC cells. Luciferase reporter plasmids were prepared to evaluate the transduction activity of $Wnt/{\beta}-catenin$ signaling pathway, and dual-luciferase reporter gene assay was arranged to confirm the targeted relationship between H19 and miR-29b-3p, as well as between miR-29b-3p and PGRN. Finally, the proliferative and invasive capacities of CRC cells were appraised through transwell, MTT and scratch assays. As a result, overexpressed H19 and down-expressed miR-29b-3p displayed close associations with the CRC patients' poor prognosis (P < 0.05). Besides, transfection with si-H19, miR-29b-3p mimic or si-PGRN were correlated with elevated E-cadherin expression, decreased snail and vimentin expressions, as well as less-motivated cell proliferation and cell metastasis (P < 0.05). Moreover, H19 was verified to directly target miR-29b-3p based on the luciferase reporter gene assay (P < 0.05), and miR-29b-3p also bound to PGRN in a direct manner (P < 0.05). Finally, addition of LiCl ($Wnt/{\beta}-catenin$ pathway activator) or XAV93920 ($Wnt/{\beta}-catenin$ pathway inhibitor) would cause remarkably altered E-cadherin, c-Myc, vimentin and snail expressions, as well as significantly changed transcriptional activity of ${\beta}-catenin/Tcf$ reporter plasmid (P < 0.05). In conclusion, the lncRNA H19/miR-29b-3p/PGRN/Wnt axis counted a great deal for seeking appropriate diagnostic biomarkers and treatment targets for CRC.

A Synthesis of Iron Oxide Based and Gadolinium Oxide Based Radiosensitizer for the Therapeutic Enhancement of Proton Beam Cancer (양성자 빔 암치료효과 개선을 위한 산화철 및 산화가돌리늄 나노입자 기반의 방사선증감제 합성)

  • Kang, Bo Sun
    • Journal of the Korean Society of Radiology
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    • v.8 no.6
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    • pp.325-332
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    • 2014
  • Metallic nanoparticles have attractive properties in biomedical applications such as diagnostics and therapeutics. Cross linked dextran coated iron oxide nanoparticles (SPIONs) and silica coated gadolinium oxide nanoparticles (SPGONs) have been synthesized as a radiosensitizer in the proton beam cancer therapy. The dextran and silicaused for the protective moieties on the SPIONs and SPGONs respectively. Size distributions of synthesized nanoparticles were confirmed 3~5 nm for SPIONs and 30~100 nm for SPGONs by transmission electron microscope (TEM). Cell survival fraction measurement and Western blot assay were performed to evaluate the radiosensitization effects of synthesized radiosensitizer. The calculated radiosensitization of SPIONs and SPGONs at 90 % cell death from the measured cell survival curves were 1.23 and 1.03 respectively. Western blotting results also show the same consistent results that the amount of released cytochrome c from mitochondria was considerably increased for the cancer cells taken up SPIONs.

Induction of Apoptotic Cell Death by Egg white combined-Chalcanthite on NCI-H460 Human Lung Cancer Cells (난담반의 인체폐암세포주 NCI-H460에 대한 세포자살유도 효능)

  • Choi, Eun-A;Kim, Kyung-Hee;Yoo, Byong-Chul;Yoo, Hwa-Seung
    • Journal of Pharmacopuncture
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    • v.12 no.3
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    • pp.49-59
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    • 2009
  • Background : Anticancer effects of herbal medicine have been reported in various types of cancer, but the systematic approaches to explain molecular mechanism(s) are not established yet. Objective : The purpose of this study is to investigate the apoptotic cell death by Egg White combined Chalcanthite in NCI-H460 human lung cancer cells. Methods : Inhibitory effects were estimated by the MTT-assay. Cancer cells were stained with DAPI and showed condensed and fragmented nuclei. The expression of cleaved caspase-3, bcl-2, and bax was detected by western blotting. To establish a basis of understanding for anti-cancer mechanism, whole proteins have been obtained from NCI-H460 harvested at 24 hrs after the treatment of Egg White combined Chalcanthite, protein expression has been profiled by 2DE-based proteomic approach. Results : NCI-H460 human lung cancer cells were treated by three samples of IS3, IS4 and IS5. IS4 inhibited most effectively the growth of NCI-H460 human lung cancer cells. The expression of cleaved caspase-3 increased in IS4 in a concentration-dependent manner. Various changes of the protein expression have been monitored, and most frequent dysregulation was found in Vimentin, Lamin-A/C. Conclusion : Egg White combined-Chacanthite inhibited the growth of NCI-H460 human lung cancer cells by inducing the apoptotic cell death via caspase-3 activation. Based upon the present findings, the further study will focus on monitoring various cancer survival factors after artificial regulation of the proteins identified, and it would be the basis for the understanding of the Chacabthite anticancer effect(s) at the molecular level.

Formulation and Cytotoxicity of Ribosome-Inactivating Protein Mirabilis Jalapa L. Nanoparticles Using Alginate-Low Viscosity Chitosan Conjugated with Anti-Epcam Antibodies in the T47D Breast Cancer Cell Line

  • Wicaksono, Psycha Anindya;Sismindari, Sismindari;Martien, Ronny;Ismail, Hilda
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.4
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    • pp.2277-2284
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    • 2016
  • Ribosome-inactivating protein (RIP) from Mirabilis jalapa L. leaves has cytotoxic effects on breast cancer cell lines but is less toxic towards normal cells. However, it can easily be degraded after administration so it needs to be formulated into nanoparticles to increase its resistance to enzymatic degradation. The objectives of this study were to develop a protein extract of M. jalapa L. leaves (RIP-MJ) incorporated into nanoparticles conjugated with Anti-EpCAM antibodies, and to determine its cytotoxicity and selectivity in the T47D breast cancer cell line. RIP-MJ was extracted from red-flowered M. jalapa L. leaves. Nanoparticles were formulated based on polyelectrolyte complexation using low viscosity chitosan and alginate, then chemically conjugated with anti-EpCAM antibody using EDAC based on carbodiimide reaction. RIP-MJ nanoparticles were characterised for the particle size, polydispersity index, zeta potential, particle morphology, and entrapment efficiency. The cytotoxicity of RIP-MJ nanoparticles against T47D and Vero cells was then determined with MTT assay. The optimal formula of RIP-MJ nanoparticles was obtained at the concentration of RIP-MJ, low viscosity chitosan and alginate respectively 0.05%, 1%, and 0.4% (m/v). RIP-MJ nanoparticles are hexagonal with high entrapment efficiency of 98.6%, average size of 130.7 nm, polydispersity index of 0.380 and zeta potential +26.33 mV. The $IC_{50}$ values of both anti-EpCAM-conjugated and non-conjugated RIP-MJ nanoparticles for T47D cells (13.3 and $14.9{\mu}g/mL$) were lower than for Vero cells (27.8 and $33.6{\mu}g/mL$). The $IC_{50}$ values of conjugated and non-conjugated RIP-MJ for both cells were much lower than $IC_{50}$ values of non-formulated RIP-MJ (>$500{\mu}g/mL$).

Apoptosis-Induced Cell Death due to Oleanolic Acid in HaCaT Keratinocyte Cells -a Proof-of-Principle Approach for Chemopreventive Drug Development

  • George, V. Cijo;Kumar, D.R. Naveen;Suresh, P.K.;Kumar, R. Ashok
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.5
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    • pp.2015-2020
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    • 2012
  • Oleanolic acid (OA) is a naturally occurring triterpenoid in food materials and is a component of the leaves and roots of Olea europaea, Viscum album L., Aralia chinensis L. and more than 120 other plant species. There are several reports validating its antitumor activity against different cancer cells apart from its hepatoprotective activity. However, antitumor activity against skin cancer has not beed studied well thus far. Hence the present study of effects of OA against HaCaT (immortalized keratinocyte) cells - a cell-based epithelial model system for toxicity/ethnopharmacology-based studies - was conducted. Radical scavenging activity ($DPPH{\cdot}$) and FRAP were determined spectrophotometrically. Proliferation was assessed by XTT assay at 24, 48 and 72 hrs with exposure to various concentrations (12.5-200 ${\mu}M$) of OA. Apoptotic induction potential of OA was demonstrated using a cellular DNA fragmentation ELISA method. Morphological studies were also carried out to elucidate its antitumor potential. The results revealed that OA induces apoptosis by altering cellular morphology as well as DNA integrity in HaCaT cells in a dose-dependent manner, with comparatively low cytotoxicity. The moderate toxicity observed in HaCaT cells, with induction of apoptosis, possibly suggests greater involvement of programmed-cell death-mediated mechanisms. We conclude that OA has relatively low toxicity and has the potential to induce apoptosis in HaCaT cells and hence provides a substantial and sound scientific basis for further validation studies.