• Title/Summary/Keyword: cell growth assay

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The Anticarcinogenic and Antioxidative Activity of Hemicentrotus pulacherrimus Fractions in Various Cancer Cells. (말똥성게 분획물에 의한 항 발암 및 항산화 효과)

  • Shin, Mi-Ok;Bae, Song-Ja
    • Journal of Life Science
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    • v.19 no.5
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    • pp.607-614
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    • 2009
  • This study was carried out to investigate the anticarcinogenic and antioxidative activities of Hemicentrotus pulacherrimus (HP). HP was extracted with methanol (HPM), which was then further fractionated into four sub-fractions by using the solvent partition method, affording methanol (HPMM), hexane (HPMH), butanol (HPMB) and aqueous (HPMA) soluble fractions. We determined the anticarcinogenic activities of these four fractions in four kinds of cancer cell lines, such as HepG2, HT29, MCF-7 and B16-F10, by MTT assay. Among various fractions from HPM, the HPMH showed the strongest growth inhibition effect. We also determined the inductive effect on quinone reductase (QR) of HP fractions. HPMB fraction exhibited strong inductive effects in HepG2 cells at a level of 90 ${\mu}g/ml$, showing inductive indexes of 2.26 compared to the control value of 1.0. The antioxidant activities of fractions from HP were also investigated by measuring the scavenging activities of HP against reactive oxygen speicies (ROS), peroxynitrite (ONOO-) and NO. Among the various solvent fractions, HPMH fractions displayed marked antioxidative activities.

Effect of Duck's Feet Derived Collagen Sponge on Skin Regeneration: In Vitro Study (오리발 유래 콜라겐 스펀지의 피부재생 효과: In vitro 연구)

  • Cha, Se Rom;Jeong, Hyun Ki;Kim, Su Young;Kim, Eun Young;Song, Jeong Eun;Park, Chan Hum;Kwon, Soon Yong;Khang, Gilson
    • Polymer(Korea)
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    • v.39 no.3
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    • pp.493-498
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    • 2015
  • For biomaterials for skin regeneration with minimized inflammatory response, high bioactivity and biocompatibility are highly required. Also, it should have a porous microstructure to improve cell adhesion and growth. In this study, we extracted a new collagen source from duck's feet which is by-product, and made the shape of sponges from duck's feet collagen (DC) to compare with DBP and SIS. To analyze physical and chemical property of the scaffold, SEM and FTIR were used. MTT assay was used to measure the attachment and proliferation of NIH/3T3 in the scaffolds. RTPCR was used to evaluate the expression of proinflammatory cytokine. Also, 1,1-diphenyl-2-picrylhydrazyl (DPPH) was used to measure the ability of antioxidant activity. Overall, this study shows that DC scaffold is biocompatible and has good physical property. Additionally, DC scaffold shows the potential as wound healing biomaterials.

Biological Activity and Manufacturing of Yanggeng with Yangha Flower Buds (양하 꽃대의 생리활성 및 양갱 제조)

  • Kim, Min-Ju;Kim, Ae-Jung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.8
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    • pp.1180-1185
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    • 2015
  • This study was performed to investigate the biological activity of yangha flower buds as well as to manufacture of yanggeng prepared with various levels (0 g, 3 g, 6 g, 9 g, and 12 g) of yangha flower buds. DPPH and ABTS scavenging activities of yangha flower buds were 96% and 57% compared to levels of vitamin C, respectively. In the oxygen radical antioxidant capacity assay, antioxidant activity increased dose dependently up to $500{\mu}g/mL$ of yangha flower buds. There was no toxicity up to $1,000{\mu}g/mL$ in vascular smooth muscle cells, and yanggeng significantly reduced migration and proliferation by platelet-derived growth factor-BB-stimulated rat aortic smooth muscle cell migration and proliferation. In the sensory evaluation, the optimal sample was YY9, which was prepared with 9 g of yangha flower buds. It can be concluded that yangha flower buds show antioxidant and vascular protective activities. The optimal sample (YY9) is expected to contribute as a new functional food.

Preparation and Characterization of Anti-GP73 Monoclonal Antibodies and Development of Double-antibody Sandwich ELISA

  • Li, Qi-Wen;Chen, Hong-Bing;Li, Zhi-Yang;Shen, Peng;Qu, Li-Li;Gong, Lai-Ling;Xu, Hong-Pan;Pang, Lu;Si, Jin
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.5
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    • pp.2043-2049
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    • 2015
  • Background: Serum Golgi protein 73 (GP73) as a novel and potential marker for diagnosing hepatocellular carcinoma (HCC) have been found to be elevated in HCC patients and associated with clinical variables representing tumor growth and invasiveness. The aim of this study was to prepare a pair of monoclonal antibodys (mAbs) against GP73 and develop a newly designed double-antibody sandwich enzyme-linked immunosorbent assay (s-ELISA), which would be used in the detection of serum GP73 (sGP73) as well as in the diagnosis of HCC. Materials and Methods: Produced by prokaryotic expression, the purified recombinant GP73 (rGP73), produced by prokaryotic expression, was used to immunize the Balb/c mice. Two hybridoma cell lines against GP73 were obtained by fusing mouse Sp2/0 myeloma cells with spleen cells from the immunized mice. The titers of anti-GP73 mAb reached 1:243,000. Western blotting analysis and Immunohistochemistry staining revealed that anti-GP73 mAb could recognize GP73 protein. The double-antibody s-ELISA was successfully established and validated by 119 HCC and 103 normal serum samples. Results: showed that the detection limit of this method could reach 1.56 ng/ml, and sGP73 levels in HCC group (mean=190.6 ng/ml) were much higher than those of in healthy controls (mean=70.92 ng/ml). Conclusions: Results of our study not only showed that sGP73 levels of HCC patients were significantly higher than those of healthy controls, but also indicated that the laboratory homemade anti-GP73 mAbs could be the optimal tool used in evaluating sGP73 levels, which would provide a solid foundation for subsequent clinical applications.

Anticancer Effect of Citrus Fruit Prepared by Gamma Irradiation of Budsticks (감귤 돌연변이체의 인간 암세포 증식 억제와 자연사멸 증강효과)

  • Kim, Ji Hye;Kim, Min Young
    • Journal of Life Science
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    • v.25 no.9
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    • pp.1051-1058
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    • 2015
  • Citrus mutant fruits were induced by irradiation of citrus budsticks with 120 Gy of cobalt (60CO) gamma irradiation. The citrus mutant inhibited the growth and induced apoptosis in various human cancer cells, including A549, HepG2, HCT116, MCF-7, and Hela. The results of a trypan blue exclusion assay showed that citrus mutant fruits exhibited excellent antiproliferation activity in various human cancer cells and low cytotoxicity in normal 16HBE140- and CHANG cells. In addition, the cell death induced by the citrus mutant fruits was associated with an increased population of cells in sub-G1 phase, and it caused DNA fragmentation in human lung adenocarcinoma A549 and hepatocellular carcinoma HepG2 cells. It also up-regulated the amount of cellular nitric oxide (NO) produced as a result of nitric oxide synthase (NOS) activation and suppressed the inhibitor of apoptosis protein (IAP) family in A549 and HepG2 cells. These findings indicate that the citrus mutant fruits activates the NO-mediated apoptotic pathway in A549 and HepG2 cells. It may merit further investigation as a potential chemotherapeutic and chemopreventive agent for the treatment of various types of cancer cells. The results provide important major new insights into the mechanisms of the anticancer activity of citrus mutant fruits.

Apoptotic Cell Death by Methanol Extract of Phellinus linteus-Barley Corn in Human Leukemic U937 Cells through Induction of p21 and Bax, and Activation of Caspase-3 (상황보리 추출물에 의한 p21 및 Bax 발현 증가와 caspase 활성화를 통한 U937 인체백혈병 세포의 apoptosis 유발)

  • Park, Cheol;Kim, Hyun-Joog;Chung, Kyung-Tae;Yoon, Tae-Kyung;Choi, Byung-Tae;Lee, Yong-Tae;Park, Dong-Il;Choi, Yung-Hyun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.5
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    • pp.1226-1232
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    • 2007
  • Phellinus linteus is a well-known Oriental medicinal fungus that has various biological activities, including immunomodulatory and anti-tumor activities, the mechanisms of which are poorly understood. In the present study, we investigated the anti-proliferative activity of the methanol extract of P. linteus-Barley corn (MEPLB) in human lekemic U937 cells. It was found that exposure of U937 cells to MEPLB resulted morphological change and growth inhibition in a dose-dependent manner as measured by trypan blue count and MTT assay. Upon treatment with MEPLB, U937 cells developed many of the hallmark features of apoptosis, including condensation of chromatin and an increase in the sub-G1 population suggesting that the anti-proliferative effect of MEPLB is associated with the induction of apoptosis. The anti-proliferative and apoptotic effects of MEPLB were connected with a marked induction of the pro-apoptotic Bax and cyclin-dependent kinase (Cdk) inhibitor p21 in a p53-independent manner. Additionally, MEPLB treatment significantly induced the caspase-3 activity in U937 cells. Taken together, the present results suggest that apoptotic signals evoked by MEPLB in human leukemic U937 cells may converge caspase-3 activation through an up-regulation of Bax rather than a down-regulation of Bcl-2 or Bel-xL.

Development of Anticancer Agents from Korean Medicinal Plants. Part 13. -Studies on the Cytotoxicity and Antitumor Activity of Herba crotalariae sessiliflorae- (한국산 생약으로부터 항암물질의 개발 (제13보). -농길리 추출물의 세포독성 및 항암작용에 관한 연구-)

  • Shin, Min-Kyo;Song, Ho-Joon;Kang, Young-Sung;Ryu, Hong-Sun;Han, Du-Seok;Kang, Kil-Ung;Baek, Seung-Hwa
    • Korean Journal of Pharmacognosy
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    • v.30 no.2
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    • pp.130-136
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    • 1999
  • The cytotoxic and antitumor activity of Herba cratalariae sessiliflorae on cultured NIH 3T3 fibroblast and human oral epitheloid carcinoma cells were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazoliumbromide (MTT) colorimetric method. The light microscopic study was carried out to observe morphological changes of cultured mouse fibroblast and human oral epitheloid carcinoma cells (KB). These results were obtained as follows; Ethyl acetate, chloroform and hexane extracts showed a significant cytotoxicity in NIH 3T3 fibroblast, but the other extracts did not show. All extracts exhibited a significant antitumor activity in human oral epitheloid carcinoma cells, but ethanol extract did not show a antitumor activity. Hexane extract showed low cytotoxic effect, but exhibited the most antitumor activity. The MTT absorbance in NIH 3T3 fibroblast was significantly decreased by treatment with chloroform, ethyl acetate and hexane extracts respectively. Human oral epitheloid carcinoma cells was significantly decreasd by treatment with all extracts with the exception of ethanol extract. The difference in MTT absorbance in two cell Types was most remarkable when treated with water and hexane extracts. Cholroform and hexane extracts showed the strongest effect in growth inhibition of human oral epitheloid carcinoma cells. These results indicated that water extract possessed no cytotoxicity and a strong antitumor activity.

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The Effect of Dohongsamul-tang on Cytokine Production in Peripheral Blood Monocular Cells of Patients with Acute Cerebral Infarction

  • Park, Sae-Wook;Lee, Sun-Woo;Kang, Baek-Gyu;Lee, In;Cho, Kwang-Ho;Moon, Byung-Soon
    • The Journal of Korean Medicine
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    • v.27 no.4
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    • pp.1-11
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    • 2006
  • Objectives : We investigated the effect of Dohongsamul-tang (DHSMT) on the production of various cytokines in lipopolysaccaride (LPS) stimulated peripheral mononuclear cells (PBMCs) from CI patients. Methods: Cell viability was determined using MTT assay. ELISA was carried out for investigating $TNF-{\alpha}$, $IL-1{\beta}$, IL-6, IL-8, IL-4, and $TGF-{\beta}$ 1 Results : The amount of tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-$1{\beta}$, IL-6, IL-8, IL-4, and transforming growth factor(TGF)-${\beta}$ 1 in PBMC culture supernatant significantly increased in the LPS treated cells compared to unstimulated cells. We show that DHSMT inhibited the production of TNF-${\alpha}$, IL-1${\beta}$, IL-6, IL-8, and IL-4 induced by LPS in a dose dependent manner. The maximal inhibition rate of the TNF-${\alpha}$, IL-1${\beta}$, IL-6, IL-8, and IL-4 production by pretreatment of DHSMT (1.0mg/ml) was 38.52 ${\pm}$ 2.5% (P < 0.01), 44.02 ${\pm}$ 3.5% (P < 0.05), 45.32 ${\pm}$ 2.3% (P < 0.01), 42.30 ${\pm}$ 3.1% (P < 0.05), and 49.70 ${\pm}$ 3.1%(P &lt; 0.05), respectively. On the other hand, DMSMT significantly increased the LPS-induced TGF-${\beta}$ 1 production(P<0.05). Conclusions : Taken together. these results suggest that DHSMT might have regulatory effects on cytokine production, which might explain its beneficial effect in the treatment of CI.

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Detection of PIK3CA Gene Mutations with HRM Analysis and Association with IGFBP-5 Expression Levels in Breast Cancer

  • Dirican, Ebubekir;Kaya, Zehra;Gullu, Gokce;Peker, Irem;Ozmen, Tolga;Gulluoglu, Bahadir M.;Kaya, Handan;Ozer, Ayse;Akkiprik, Mustafa
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.21
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    • pp.9327-9333
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    • 2014
  • Breast cancer is the second most common cancer and second leading cause of cancer deaths in women. Phosphatidylinositol-3-kinase (PI3K)/AKT pathway mutations are associated with cancer and phosphatidylinositol-4, 5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) gene mutations have been observed in 25-45% of breast cancer samples. Insulin growth factor binding protein-5 (IGFBP-5) can show different effects on apoptosis, cell motility and survival in breast cancer. We here aimed to determine the association between PIK3CA gene mutations and IGFBP-5 expressions for the first time in breast cancer patients. Frozen tumor samples from 101 Turkish breast cancer patients were analyzed with high resolution melting (HRM) for PIK3CA mutations (exon 9 and exon 20) and 37 HRM positive tumor samples were analyzed by DNA sequencing, mutations being found in 31. PIK3CA exon 9 mutations (Q546R, E542Q, E545K, E542K and E545D) were found in 10 tumor samples, exon 20 mutations (H1047L, H1047R, T1025T and G1049R) in 21, where only 1 tumor sample had two exon 20 mutations (T1025T and H1047R). Moreover, we detected one sample with both exon 9 (E542Q) and exon 20 (H1047R) mutations. 35% of the tumor samples with high IGFBP-5 mRNA expression and 29.4% of the tumor samples with low IGFBP-5 mRNA expression had PIK3CA mutations (p=0.9924). This is the first study of PIK3CA mutation screening results in Turkish breast cancer population using HRM analysis. This approach appears to be a very effective and reliable screening method for the PIK3CA exon 9 and 20 mutation detection. Further analysis with a greater number of samples is needed to clarify association between PIK3CA gene mutations and IGFBP-5 mRNA expression, and also clinical outcome in breast cancer patients.

Caffeic Acid Phenethyl Ester Induces the Expression of NAG-1 via Activating Transcription Factor 3 (ATF3를 통한 caffeic acid phenethyl ester에 의한 NAG-1 유전자의 발현 증가)

  • Park, Min-Hee;Chung, Chungwook;Lee, Seong Ho;Baek, Seung Joon;Kim, Jong Sik
    • Journal of Life Science
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    • v.28 no.1
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    • pp.37-42
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    • 2018
  • Non-steroidal anti-inflammatory drug-activated gene-1 (NAG-1) is a transforming growth factor beta (TGF-${\beta}$) superfamily gene associated with pro-apoptotic and anti-tumorigenic activities. In the present study, we investigated if caffeic acid phenethyl ester (CAPE) derived from propolis could induce the expression of anti-tumorigenic gene NAG-1. Our results indicate that CAPE significantly induced NAG-1 expression in a time- and concentration-dependent manner in HCT116 cells. We also found that CAPE induced NAG-1 expression in a concentration-dependent manner in another human colorectal cancer cell line, LOVO. In addition, CAPE triggered apoptosis, which was detected with Western blot analysis using poly-(ADP-ribose) polymerase antibody. NAG-1 induction by CAPE was not dependent on transcription factor p53, which was confirmed with Western blot analysis using p53 null HCT116 cells. The luciferase assay results indicated that the new cis-elements candidates were located between -474 and -1,086 of the NAG-1 gene promoter. CAPE dramatically induced activating transcription factor 3 (ATF3) expression, but not cAMP response element-binding protein (CREB), which shares the same binding sites with ATF3. The co-transfection experiment with pCG-ATF3 and pCREB showed that only ATF3 was associated with NAG-1 up-regulation by CAPE, whereas CREB had no effect. In conclusion, the results suggest that CAPE could induce the expression of anti-tumorigenic gene NAG-1 mainly through ATF3.