• Title/Summary/Keyword: MTT assay

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Inhibitory Effect of Ginseng on Breast Cancer Cell Line Growth Via Up-Regulation of Cyclin Dependent Kinase Inhibitor, p21 and p53

  • Shabanah, Othman A AL;Alotaibi, Moureq R;Rejaie, Salim S Al;Alhoshani, Ali R;Almutairi, Mashal M;Alshammari, Musaad A;Hafez, Mohamed M
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.11
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    • pp.4965-4971
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    • 2016
  • Objective: Breast cancer is global female health problem worldwide. Most of the currently used agents for breast cancer treatment have toxic side-effects. Ginseng root, an oriental medicine, has many health benefits and may exhibit direct anti-cancer properties. This study was performed to assess the effects of ginseng on breast cancer cell lines. Materials and Methods: Cytotoxicity of ginseng extract was measured by MTT assay after exposure of MDA-MB-231, MCF-10A and MCF-7 breast cancer cells to concentrations of 0.25, 0.5, 1, 1.5, 2 and 2.5 mg/well. Expression levels of p21WAF, p16INK4A, Bcl-2, Bax and P53 genes were analyzed by quantitative real time PCR. Results: The treatment resulted in inhibition of cell proliferation in a dose-and time-dependent manner. p53, p21WAF1and p16INK4A expression levels were up-regulated in ginseng treated MDA-MB-231 and MCF-7 cancer cells compared to untreated controls and in MCF-10A cells. The expression levels of Bcl2 in the MDA-MB-231 and MCF-7 cells were down-regulated. In contrast, that of Bax was significantly up-regulated. Conclusion: The results of this study revealed that ginseng may inhibit breast cancer cell growth by activation of the apoptotic pathway.

Cytotoxicity of Diesel Exhaust Particles from Various Vehicles toward Macrophage Cells (국내 디젤 차량 배기 입자가 쥐 대식세포에 미치는 세포독성 평가)

  • Lee, Jang-Han;Lee, Yong-Kwon;Lee, Ji-Young;Lee, Seung-Bok;Kim, Sun-Hwa;Bae, Gwi-Nam;Lee, Hak-Sung;Lim, Cheol-Soo;Chung, Nam-Hyun
    • Environmental Analysis Health and Toxicology
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    • v.25 no.2
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    • pp.111-120
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    • 2010
  • DEPs (diesel exhaust particles) like any other particles can be also inhaled into lung to participate in a damaging reaction to the organ. Possible damages might be apoptosis and inflammatory responses to the cells in respiratory track. The aim of this study was cytotoxicity evaluation of DEPs from five in-use diesel vehicles using a murine macrophage cell (RAW 254.7). We found that most DEPs have a considerable cytotoxicity compared to the control and SRM 2975. When measured by MTT assay and extents of apoptosis, DEPs of two highmileage vehicles had higher toxicity than those of the other three low-mileage vehicles tested. Although mRNA expression level of TNF-${\alpha$ somewhat explains the trend of cytotoxicity and apoptosis, that of IL-1$\beta$ did not. Correlation studies among the extents of MTT assay, apoptosis, and TNF-$\alpha$ expression showed that the extents between apoptosis and TNF-$\alpha$ expression was most highly correlated (r=0.96). These results suggest that cytotoxicity of various DEPs could be compared easily by measuring the extent of apoptosis or TNF-$\alpha$ expression by DEPs.

Effect of Red Ginseng Extract on the Inflammatory Response of LPS-Induced RAW 264.7 cell (홍삼추출물이 LPS로 유도된 RAW 264.7 cell의 염증반응에 미치는 효과)

  • Jang, Young-Ah;Kim, Han-Na;Kim, Bo-Ae
    • Journal of the Korean Applied Science and Technology
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    • v.36 no.4
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    • pp.1434-1442
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    • 2019
  • We conducted this study to investigate anti-inflammatory possibilities of applying cosmetic material about extracts from red ginseng. For this we carried out biological active evaluation about anti-inflammatory by using extracts of red ginseng. In order to evaluate the anti-inflammatory effects of the samples in macrophages (RAW 264.7 cells), MTT assay was used to evaluate the toxicity of red ginseng extracts and the inhibitory activity of nitric oxide production and the expression levels of inflammation-related proteins and genes. The inhibitory activity of nitric oxide in the LPS-induced RAW 264.7 cells was 71.2% at 25 ㎍/ml concentration and western blot analysis showed that the expression of iNOS and COX-2 protein decreased in a concentration-dependent manner. These results suggest that extracts from red ginseng may have value as the potential cosmetic materials.

The Antioxidation Effect of Salsola komarovii Extract and Its Influence on Cell Bio activity (수송나물(Salsola komarovii)의 항산화, 항염 및 미백 활성)

  • Kim, Min Jeong;Kim, Sook-Hee;Lee, Ja-Bok
    • Journal of Convergence for Information Technology
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    • v.10 no.10
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    • pp.308-317
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    • 2020
  • S. komarovii is halophyte that grows in soil or waters of high salinity, such as in saline semi-deserts, sloughs and seashores. Traditionally, S. komarovii has been used for food and medicinal purposes in Korea. S. komarovii was extracted in 70% ethanol to measure anti-oxidative activity using DPPH and ABTS assay. The IC50 values of the S. komarovii extract against DPPH radicals and ABTS radicals were 186.10 mg/mL and 121.89 mg/mL. In addition, total polyphenol and reducing power were measured. The S. komarovii extract exhibited superior polyphenolic (22.5%) and antioxidant (28.4%) contents. Regarding cell bioactivity, MTT assay was conducted to reveal cytotoxicity of S. komarovii extract and showed the non-cytotoxicity of S. komarovii extract. Anti-inflammatory and skin whitening effects were measured at 100 ㎍/mL. Therefore, this study suggests that the S. komarovii extract can be used as a functional cosmetic product material.

Antiproliferative effect of Chungjogupae-tang treatment was associated with the inhibition of prostaglandin E2 release and Telomere active in human lung carcinoma cells (인체폐암세포에서의 prostaglandin E2 생성과 Telomere 활성에 미치는 청조구폐탕의 영향에 관한 연구)

  • Kim, Hoon;Park, Dong-Il
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.19 no.2
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    • pp.26-39
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    • 2006
  • Objective : The effect of water extract of Chungjogupae-tang (CJGPT) was investigated on the growth of human lung carcinoma A549 cells. Methods : MTT assay and fluorescent microscope performed to compare and examine the efficacy of CJGPT treatment on the cytostaticity of lung cancer cells in proportion to time and doses, and DAPI staining and Western blot analysis were used to examine their effect on apoptosis. In addition the quantitative RT-PCR was used to examine to lung cancer cells growth and Progtaglandin E2 and Telomerase activity were measured Results : Exposure of A549 cells to CJGPT resulted in the growth inhibition and apoptosis in a dose-dependent manner as measured by MTT assay and fluorescent microscope. The antiuoliferative effect by CJGPT treatment in A549 cells was associated with morphological changes such as membrane shrinking and cell rounding up. CJGPT treatment resulted in an up-regulation of cyclin-dependent kinase inhibitor p21(WAF1/CIPl) in a p53-independent fashion. We found that CJGPT treatment decreased the levels of cyclooxygenase (COX)-2 and inducible nitric oxide synthease (iNOS) expression without significant changes in the expression of COX-1, which was correlated with a decrease in protaglandin E2 (PGE2) synthesis. CJGPT treatment also inhibited the levels of human telomerase reverse transcriptase (hTERT) and telomerase-associated protein (TEP)-1 mRNA expression, however the activity of telomerase was slightly increased by CJGPT treatment. Conclusion : These findings suggested that CJGPT-induced inhibition of human lung carcinoma A549 cell growth was connected with the induction of apoptotic cell death and the results provided important new insights into the possible molecular mechanisms of the anti-cancer activity of CJGPT.

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Chemical composition, antioxidant potential and cyto-protecting activity of essential oil of Liriodendron tulipifera L. leaves

  • Yadav, Anil Kumar;Kim, Sang Ho;Kang, Sun Chul
    • The Korea Journal of Herbology
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    • v.30 no.4
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    • pp.1-9
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    • 2015
  • Objectives : The present study was under taken to characterize chemical composition, antioxidant and cyto-protecting capacity of essential oil obtained from leaves of Liriodendron tulipifera L. Methods : Essential oil from the leafof L. tulipifera L. (EOLL) was extracted by hydro-distillation process and further its chemical composition was evaluated by GC-MS analysis. The in vitro antioxidant potential of the EOLL was determined by DPPH , ABTS ●+, superoxide and nitric oxide free radical scavenging activity using different concentrations in the range of 50-800 μg/mL. In addition, cyto-protecting property of the EOLLwas determined by MTT assay on Raw 264.7 macrophage cells challenged with hydrogen peroxide (H 2 O 2 ). Results : The result of GC-MS analysis showed presence of 34 volatile compounds, principally germacrene D, spathulenol, and α -cadinol in EOLL. The in vitro antioxidant assays of EOLL at the highest used concentration of 800 μg/mL showed 81.62, 84.29, 83.59 and 58.59% inhibition of DPPH , ABTS ●+, superoxide, and nitric oxide radicals, respectively. It also showed ferric reducing ability with 1310.04 mM Fe (II)/g of essential oil. The EOLL at three different concentrations (200, 400 and 800 μg/mL) protected the cells from H 2 O 2 -induced cell damage through scavenging intracellular ROS. Conclusion : The findings from the study suggest that essential oil isolated from leaves of L tulipifera L. is a potent sources of natural antioxidants, which could be used to treat the diseases associated with oxidative stress condition.

CHARACTERISTICS OF FIBROUS DYSPLASIA DERIVED CELLS (섬유성이형성증 유래세포의 특성연구)

  • Lee, Chan-Hee;Han, Ihn;Seo, Byoung-Moo
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.35 no.5
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    • pp.304-309
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    • 2009
  • Purpose: Fibrous dysplasia (FD) is a fibro-osseous disease associated with activating missense mutations of the gene encoding the $\alpha$-subunit of stimulatory G protein. FD may affect a single bone (called monostotic form) or multiple bones (called polyostotic form). The extent of lesions reflects the onset time of mutation. In this study, cells from monostotic FD in maxilla of a patient were isolated and cultured in vitro for characterization. Materials and Methods: The single cells were released from FD lesion which was surgical specimen from 15 years-old boy. These isolated cells were cultured in vitro and tested their proliferation activity with MTT assay. In osteogenic media, these cells underwent differentiation process comparing with its normal counterpart i.e. bone marrow stromal cells. The proliferated FD cells were detached and transplanted into the dordsal pocket of nude mouse and harvested in 6 weeks and 12 weeks. Results and Summary: FD cells have an increased proliferation rate and poor differentiation. As a result, cells isolated from FD lesion decreased differentiation into osteoblast and increased proliferation capacity. MTT assay presented that proliferation rate of FD cells were higher than control. However, the mineral induction capacity of FD was lesser than that of control. Monostotic FD cells make fewer amounts of bone ossicles and most of them are woven bone rather than lamellar bone in vivo transplantation. In transplanted FD cells, hematopoietic marrow were not seen in the marrow space and filled with the organized fibrous tissue. Therefore, they were recapitulated to the original histological features of FD lesion. Collectively, these results indicated that the FD cells were shown that the increased proliferation and decreased differentiation potential. These in vitro and in vivo system can be useful to test FD cell's fate and possible.

Cytotoxicity Assessment of Six Different Extracts of Abelia triflora leaves on A-549 Human Lung Adenocarcinoma Cells

  • Al-Taweel, Areej Mohammad;Perveen, Shagufta;Fawzy, Ghada Ahmed;Ibrahim, Taghreed Abdou;Khan, Afsar;Mehmood, Rashad
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.11
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    • pp.4641-4645
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    • 2015
  • The present investigation was designed to assess the anticancer activity of six different leaf extracts (ethyl acetate, methanol, chloroform, petroleum ether, n-butanol, and water soluble) of Abelia triflora on A-549 human lung adenocarcinoma epithelial cells. A-549 cells were exposed to $10-1000{\mu}g/ml$ concentrations of the leaf extracts of A. triflorafor 24 h and then percentage cell viability was assessed by 3-(4,5-dimethylthiazol-2yl)-2,5-biphenyl tetrazolium bromide (MTT) assay. The results showed that leaf extracts of A. triflora significantly reduced the viability of A-549 cells in a concentration-dependent manner. Decrease was recorded as 31% with ethyl acetate, 36% with methanol, 46% with chloroform, 54% with petroleum ether, 62% with n-butanol, and 63% with water soluble extracts at $1000{\mu}g/ml$ each. Among the various plant extracts, ethyl acetate extract showed the highest decrease in the percentage cell viability, followed by methanol, chloroform, petroleum ether, n-butanol, and water soluble extracts. Our results demonstrated preliminary screening of anticancer activity of different soluble extracts of A. triflora extracts against A-549 cells, which can be further used for the development of a potential therapeutic anticancer agents.

CELLULAR ATTACHMENT AND GENE EXPRESSION OF OSTEOBLAST-LIKE CELLS ON ZIRCONIA CERAMIC SURFACES

  • Pae, Ah-Ran;Lee, Hee-Su;Kim, Hyeong-Seob; Baik, Jin;Woo, Yi-Hyung
    • The Journal of Korean Academy of Prosthodontics
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    • v.46 no.3
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    • pp.227-237
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    • 2008
  • STATEMENT OF PROBLEM: Zirconium oxide can be a substitute to titanium as implant materials to solve the esthetic problems of dark color in the gingival portion of implant restorations. PURPOSE: This study was performed to define attachment and growth behavior of osteoblast- like cells cultured on grooved surfaces of zirconium oxide and evaluate the genetic effect of zirconium oxide surfaces using the reverse transcriptase-polymerase chain reaction (RT-PCR). MATERIAL AND METHODS: MC3T3-E1 cells were cultured on (1) commercially pure titanium discs with smooth surface (T group), (2) yttrium-stabilized tetragonal zirconia polycrystal (Y-TZP) with machined surface (ZS group), and (3) Y-TZP with $100{\mu}m$ grooves (ZG group). Cell proliferation activity was evaluated through MTT assay and cell morphology was examined by SEM. The mRNA expression of Runx2, alkaline phosphatase, osteocalcin, TGF-${\beta}1$, IGF-1, G3PDH in E1 cells were evaluated by RT-PCR. RESULTS: From the MTT assay, after 48 hours of adhesion of MC3T3-E1 cells, the mean optical density value of T group and ZG group significantly increased compared to the ZS group. SEM images of osteoblast-like cells showed that significantly more cells were observed to attach to the grooves and appeared to follow the direction of the grooves. After 24 hours of cell adhesion, more spreading and flattening of cells with active filopodia formation occurred. Results of RT-PCR suggest that T group, ZS group, and ZG group showed comparable osteoblast-specific gene expression after 24 hours of cell incubation. CONCLUSION: Surface topography and material of implants can play an important role in expression of osteoblast phenotype markers. Zirconia ceramic showed comparable biological responses of osteoblast-like cells with titanium during a short-time cell culture period. Also, grooves influence cell spreading and guide the cells to be aligned within surface grooves.

Experimental Study on the Anti-depressive Effect of Bee Venom Injection (봉독이 우울증 모델 흰쥐에게 미치는 영향)

  • Lee, Jin-Hee;Kim, Geun-Woo;Koo, Byung-Soo
    • Journal of Oriental Neuropsychiatry
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    • v.21 no.2
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    • pp.61-73
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    • 2010
  • Objectives : The purpose of this study was to assess anti-depressive effects of Bee Venom(BV) on an Animal Model of Depression induced immobility stress. Methods : There was 2 pre-experiments MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and Western blot test and 3 main experiments ; forced swimming test, tail suspension test and Y-maze task. Male rats were used for main experiment. The subject was divided into 4 groups(1. control group injected only saline, without immobility stress 2. Negative group injected saline after 2 hours immobility stress 3. Positive group injected Amitriptyline after 2 hours immobility stress 4. BV group injected Bee Venom after 2 hours immobility stress). Each group consisted of 6 rats. Forced swimming test, tail suspension test, Y-maze task were used to evaluate anti-depressive effect of Bee Venom. Results : In MTT assay, as the density of BV increased, the existence rate of primary neuronal cell increased. In Western blot test, the density of CREB and AKT was increasing as time went by. In forced swimming test, BV group showed immobility decreased more than Normal group and Positive group. In tail suspension test, Normal group and Positive group showed immobility decreased more than BV group. In Y-maze task, BV group showed immobility decreased more than Normal group, but Positive group showed immobility decreased more than BV group. Conclusions : These results suggest that Bee Venom may have anti-depressive effect on depression.