• Title/Summary/Keyword: MTT assay

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An Experimental Study on the Effect of Angelica acutiloba Ethanol Extract on Hyaluronic Acid Synthesis (일당귀 에탄올 추출물의 Hyaluronic Acid 합성 효과에 대한 실험적 연구)

  • Kang, Min-Seo;Ha, Hun-Yong;Kim, Hee-Taek
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.28 no.1
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    • pp.32-40
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    • 2015
  • Objectives : Hyaluronic acid(HA) is a mucopolysaccharide, occuring naturally in living organisms. It is one of the most hydrophilic molecules, so it has been known as being related to skin hydration and anti-aging. The purpose of this study is to examine the effects of Angelica acutiloba ethanol extract on hyaluronic acid synthesis. Methods : To determine cytotoxicity and hyaluronic acid synthase 2 gene expression, hyaluronic acid production in HaCaT cells, MTT assay and RT-PCR ELISA was used. Results : There was no cytotoxicity in $50{\mu}g/ml$ concentration Angelica acutiloba extract in MTT assay. Hyaluronic acid synthase 2(HAS2) gene expression was increased by all treated concentration Angelica acutiloba extract. Hyaluronic acid production was higher in $50{\mu}g/ml$ & $100{\mu}g/ml$ concentration Angelica acutiloba extract than control group. Conclusions : Hyaluronic acid production was increased by Angelica Acutiloba extracts. Therefore, We suggest that Angelica acutiloba can make a contribution to the moisturing effect on human skin. Conclusions : Hyaluronic acid production was increased by Angelica Acutiloba extracts. Therefore, We suggest that Angelica acutiloba can make a contribution to the moisturing effect on human skin.

Combination Effect of Nimotuzumab with Radiation in Colorectal Cancer Cells (대장암 세포에서 EGFR 저해제 Nimotuzumab의 방사선 병합 효과)

  • Shin, Hye-Kyung;Kim, Mi-Sook;Jeong, Jae-Hoon
    • Radiation Oncology Journal
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    • v.28 no.3
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    • pp.147-154
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    • 2010
  • Purpose: To investigate the radiosensitizing effect of the selective epidermal growth factor receptor (EGFR) inhibitor nimotuzumab in human colorectal cancer cell lines. Materials and Methods: Four human colorectal cancer cell lines, HCT-8, LoVo, WiDr, and HCT-116 were treated with nimotuzumab and/or radiation. The effects on cell proliferation, viability, and cell cycle progression were measured by MTT, clonogenic survival assay, flow cytometry, and Western blot. Results: An immunoblot analysis revealed that EGFR phosphorylation was inhibited by nimotuzumab in colorectal cancer cell lines. Under these experimental conditions, pre-treatment with nimotuzumab increased radiosensitivity of colorectal cancer cell lines, except for cell line HCT-116. However, cell proliferation or cell cycle progression was not affected by the addition of nimotuzumab, irrespective of irradiation. Conclusion: Nimotuzumab enhanced the radiosensitivity of colorectal cancer cells in vitro by inhibiting EGFR-mediated cell survival signaling. This study provided a rationale for the clinical application of the selective EGFR inhibitor, nimotuzumab in combination with radiation in colorectal cancer cells.

Effects of Garlic Oil on Pancreatic Cancer Cells

  • Lan, X.Y.;Sun, H.Y.;Liu, J.J.;Lin, Y.;Zhu, Z.Y.;Han, X.;Sun, X.;Li, X.R.;Zhang, H.C.;Tang, Z.Y.
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.5905-5910
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    • 2013
  • Background: To investigate the preventive and therapeutic potential of garlic oil on human pancreatic carcinoma cells. Methods: The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay was performed to study the effects of garlic oil on three human pancreatic cancer cell lines, AsPC-1, Mia PaCa-2 and PANC-1. Cell cycle progression and apoptosis were detected by flow cytometry (FCM), staining with PI and annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI), respectively. Morphologic changes of pancreatic cancer cells were observed under transmission electron microscopy (TEM) after treatment with garlic oil at low inhibitory concentrations ($2.5{\mu}M$ and $10{\mu}M$) for 24 hours. Results: Proliferation of the AsPC-1, PANC-1, and Mia PaCa-2 cells was obviously inhibited in the first 24 hours with the MTT assay. The inhibition effect was more significant after 48 hours. When cells were exposed to garlic oil at higher concentrations, an early change of the apoptotic tendency was detected by FCM and TEM. Conclusion: Garlic oil could inhibit the proliferation of AsPC-1, PANC-1, and Mia PaCa-2 cells in this study. Moreover, due to programmed cell death, cell cycle arrest, or both, pro-apoptosis effects on AsPC-1 cells were induced by garlic oil in a dose and time dependent manner in vitro.

Inhibition of Tumor Growth in Vitro by a Combination of Extracts from Rosa Roxburghii Tratt and Fagopyrum Cymosum

  • Liu, Wei;Li, Su-Yi;Huang, Xin-En;Cui, Jiu-Jie;Zhao, Ting;Zhang, Hua
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.5
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    • pp.2409-2414
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    • 2012
  • Objective: Traditional Chinese herbal medicines have a very long history. Rosa roxburghii Tratt and Fagopyrum cymosum are two examples of plants which are reputed to have benefits in improving immune responses, enhancing digestive ability and demonstrating anti-aging effects. Some evidence indicates that herbal medicine soups containing extracts from the two in combination have efficacy in treating malignant tumors. However, the underlying mechanisms are far from well understood. The present study was therefore undertaken to evaluate anticancer effects and explore molecular mechanisms in vitro. Methods: Proliferation and apoptosis were assessed with three carcinoma cell lines (human esophageal squamous carcinoma CaEs-17, human gastric carcinoma SGC-7901 and pulmonary carcinoma A549) by MTT assay and flow cytometry, respectively, after exposure to extract from Rosa roxburghii Tratt (CL) and extract from Fagopyrum cymosum (FR). $IC_{30}$ of CL and FR were obtained by MTT assay. Tumor cells were divided into four groups : control with no exposure to CL or FR; CL with $IC_{30}$ CL; FR with $IC_{30}$ FR; CL+FR group with 1/2 ($IC_{30}$ CL + $IC_{30}$ FR). RT-PCR and Western blot analysis were used to detect the expression of Ki-67, Bax and Bcl-2 at mRNA and protein levels. Results: Compared with the CL or FR groups, the combination of CL+FR showed significant inhibition of cell growth and increase in apoptosis; the mRNA and protein expression levels of Ki-67 and Bcl-2 in CL+FR group were all greatly decreased, while the expression of Bax was markedly increased. Conclusions: These results indicate that the synergistic antitumor effects of combination of CL and FR are related to inhibition of proliferation and induction of apoptosis.

Steroidal Saponins from Paris polyphylla Suppress Adhesion, Migration and Invasion of Human Lung Cancer A549 Cells Via Down-Regulating MMP-2 and MMP-9

  • He, Hao;Zheng, Lei;Sun, Yan-Ping;Zhang, Guang-Wei;Yue, Zheng-Gang
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.24
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    • pp.10911-10916
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    • 2015
  • Background: Tumor metastases are the main reasons for oncotherapy failure. Paris polyphylla (Chinese name: Chonglou) has traditionally been used for its anti-cancer actions. In this article, we focus on the regulation of human lung cancer A549 cell metastases and invasion by Paris polyphylla steroidal saponins (PPSS). Materials and Methods: Cell viability was evaluated in A549 cells by MTT assay. Effects of PPSS on invasion and migration were investigated by wound-healing and matrigel invasion chamber assays. Adhesion to type IV collagen and laminin was evaluated by MTT assay. Expression and protease activity of two matrix metalloproteinases (MMPs), MMP-2 and MMP-9, were analyzed by Western blotting and gelatin zymography, respectively. Results: PPSS exerted growth inhibitory effects on A549 cells, and effectively inhibited A549 cell adhesion, migration and invasion in a concentration-dependent manner. Western blotting and gelatin zymography analysis revealed that PPSS inhibited the expression and secretion of MMP-2 and MMP-9 in A549 cells. Conclusions: PPSS has the potential to suppress the migration, adhesion and invasion of A549 cells. PPSS could be a potential candidate for interventions against lung cancer metastases.

The Effects on Antimicrobial and Anticarcinogenic Activity of Momordica Charantia L. (메탄올로 추출한 여주 분획성분의 항균 및 항발암 효과)

  • 배송자
    • Journal of Nutrition and Health
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    • v.35 no.8
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    • pp.880-885
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    • 2002
  • This study was performed to determine the antimicrobial and anticarcinogenic activities of the Momordica charantia L. (MC) on several microorganisms and human cancer cell lines. In the paper disk test, its antimicrobial activity was increased in proportion to its concentration. Among the various solvent fractions of Momordica charantia L., the ethylether partition layer (MCMEE) showed the strongest antimicrobial activity. Also, the ethylacetate partition layer (MCMEA) and the butanol partition layer (MCMB) showed antimicrobial activity. We also determined the cytotoxicity and chemopreventive effect of Momordica charantia L. extract and fractions on human cancer cells. The experiment was conducted to determine the cytotoxicity of Momordica charantia L. partition layers on HepG2, HeLa and MCF-7 cells by MTT assay. Among the various partition layers of Momordica charantia L., MCMEE and MCMEA showed strong cytotoxic effects on all cancer cell lines. The chemopreventive effect of the quinone reductase induced activities of HepG2 cell, the hexane partition layer (MCMH) at a dose of 50 $\mu\textrm{g}$/mL was 3.62 times more effective compared with the control values of 1.0. Therefore, based on these studies, Momordica charantia L. may be developed into a potentially useful cancer chemopreventive agent.

제 3세대 백금착체 항암제 신약개발 2. Antitumor activity and ex vivo pharmacodynamics of SKI 2053R

  • 박재갑;홍원선;방영주;조용백;태주호;김훈택;김대기;김기협;김노경
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.74-74
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    • 1993
  • The in vitro cytotoxicity of SKI 2053R was evaluated against human tumor cell lines along with those of cisplatin and carboplatin using MTT assay. The cell lines tested were two human lung cancer cell lines and five human stomach cancer celt lines. The level of cytotoxic effects of SKI 2053R against two human lung cancer cell lines was located between cisplatin and carboplatin. However, the cytotoxic activity of SKI 2053R against five human stomach cancer cell lines was similar to that of cisplatin. SKI 2053R is considered to be selectively cytotoxic toward human stomach cancer cell lines. We carried out pharmacokinetic and ex vivo phrmacodynamic studies of SKI 2053R in beagle dogs to predict the clinical antitumor effect of SKI2053R, comparing with those of cisplatin and carboplatin. In ex vivo pharmacodynamics which used MTT assay as bioassay on the 2 lung and 5 stomach cancer cell, mean antitumor indexes (ATIs) of SKI 2053R were highest among three compounds in both lung and stomach cancer cell lines, especially in stomach cancer cell. Much higher ATI profile and maximal inhibition rates of SKI 2053R appeared in the stomach cancer cells will give desirable advantages to clinical trial s against gastric carcinoma. The anti tumor activity and target organ toxicity of SKI 2053R were compared with those of cisplatin on stomach cancer cell line, KATO III xenografted into nude BALB/c(nu/nu) mice. All groups of cisplatin and SKI 2053R showed active tumor regression. The inhibition rates(IR) of SKI 2053R were higher than that of cisplatin on the basis of mean IR. Though the loss of body weight was observed in all groups from the first week, the SKI 2053R group recovered it soon from the third week after the initiation of treatment, maintaining the most active anti tumor activity among three groups.

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The Effects on Antimicrobial and Cytotoxicity of Brassica oleracea L. Fractions. (적채 분획물의 항균, 암세포 증식 억제효과)

  • 박윤자;전광혜;김선희;배송자
    • Journal of Life Science
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    • v.14 no.4
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    • pp.567-572
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    • 2004
  • This study was carried out to investigate the affects on antimicrobial and cytotoxicity of red cabbage (Brassica oleracea L.,BO). In the paper disc test, antimicrobial activity of BO fractions was increased in proportion to its concentration. Among the various solvent fractions of methanol extract (BOM) of BO, the ethylacetate partition layer (BOMEA) showed the strongest antimicrobial activity We also determined the cytotoxicity and Quinone Reductase induced effect of BO extract and fractions on human cancer cells. The cytotoxicity of BO fractions on HepG2, HeLa and MCF-7 cells was evaluated by MTT assay. The BOMEE and BOMEA showed strong cytotoxic effects on all cancer cell lines we used. The quinone reductase induced effect of BO fractions on HepG2 cells, the hexane partition layer (BOMH) at a dose of 200 $\mu{g}$/ml was 2.88 times more effective compared to the control values of 1.0.

In vitro cytotoxicity of four calcium silicate-based endodontic cements on human monocytes, a colorimetric MTT assay

  • Khedmat, Sedigheh;Dehghan, Somayyeh;Hadjati, Jamshid;Masoumi, Farimah;Nekoofar, Mohammad Hossein;Dummer, Paul Michael Howell
    • Restorative Dentistry and Endodontics
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    • v.39 no.3
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    • pp.149-154
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    • 2014
  • Objectives: This study was performed to evaluate the cytotoxicity of four calcium silicate-based endodontic cements at different storage times after mixing. Materials and Methods: Capillary tubes were filled with Biodentine (Septodont), Calcium Enriched Mixture (CEM cement, BioniqueDent), Tech Biosealer Endo (Tech Biosealer) and ProRoot MTA (Dentsply Tulsa Dental). Empty tubes and tubes containing Dycal were used as negative and positive control groups respectively. Filled capillary tubes were kept in 0.2 mL microtubes and incubated at $37^{\circ}C$. Each material was divided into 3 groups for testing at intervals of 24 hr, 7 day and 28 day after mixing. Human monocytes were isolated from peripheral blood mononuclear cells and cocultered with 24 hr, 7 day and 28 day samples of different materials for 24 and 48 hr. Cell viability was evaluated using an MTT assay. Results: In all groups, the viability of monocytes significantly improved with increasing storage time regardless of the incubation time (p < 0.001). After 24 hr of incubation, there was no significant difference between the materials regarding monocyte viability. However, at 48 hr of incubation, ProRoot MTA and Biodentine were less cytotoxic than CEM cement and Biosealer (p < 0.01). Conclusions: Biodentine and ProRoot MTA had similar biocompatibility. Mixing ProRoot MTA with PBS in place of distilled water had no effect on its biocompatibility. Biosealer and CEM cement after 48 hr of incubation were significantly more cytotoxic to on monocyte cells compared to ProRoot MTA and Biodentine.

Secretion of Inflammatory Cytokines by Aloe vera Extract in HepG2 Cells (HepG2 세포에서 알로에 베라 추출물에 의한 염증성 사이토카인 분비)

  • Kim, Ilrang
    • The Korean Journal of Food And Nutrition
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    • v.27 no.3
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    • pp.400-405
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    • 2014
  • Recently, cases of Aloe vera induced-toxic hepatitis have been reported. However, the precise inflammatory effects of Aloe vera extract have not been clearly elucidated yet. In this study, the inflammatory effects and the mechanism of 70% ethanolic Aloe vera extract on liver were evaluated by in vitro assays using human hepatoma HepG2 cells. Cell viability was investigated using MTT assay at $0.001{\sim}100{\mu}g/mL$ of Aloe vera extract. To evaluate inflammatory effect of the Aloe vera extract, the secretion of pro-inflammatory cytokine Interleukin 8 (IL-8) and Macrophage colony-stimulating factor (M-CSF) were detected. Aloe vera extract did not induce cell death at concentrations of $0.001{\sim}100{\mu}g/mL$. However, Aloe vera extract significantly increased the IL-8 secretion by 15.7~25.8% and the M-CSF secretion by 36.6~61.5% at the same concentrations. These results indicate that Aloe vera extract shows an inflammation-related mild hepatotoxicity than a severe toxicity such as cell death and this hepatitis is mediated by the secretion of inflammatory cytokine IL-8 and M-CSF.