• Title/Summary/Keyword: cell growth assay

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Inhibitory Effect of Linum usitatissimum and Perilla frutescens as Sources of Omega-3 Fatty Acids on Mutagenicity and Growth of Human Cancer Cell Lines (식물성 오메가-3계 지방산 급원인 아마씨 및 들깨의 항돌연변이 및 암세포 증식 억제 효과)

  • Lim, Sun-Young
    • Journal of Life Science
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    • v.19 no.12
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    • pp.1737-1742
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    • 2009
  • It has been known that Linum usitatissimum and Perilla frutescens are dietary sources of possible chemopreventive compounds such as lignans and $\alpha$-linolenic acid. Here, we investigated and compared the inhibitory effects of methanol extracts from Linum usitatissimum and Perilla frutescens on mutagenicity using the Ames test, and growth of human cancer cells (AGS human gastric adenocarcinoma, HT-29 human colon cancer, Hep 3B hepatocellular carcinoma cells). In the Ames test system using Salmonella typhimurium TA100, aflatoxin $B_1$ ($AFB_1$)-induced mutagenicity was significantly inhibited by treatment with the methanol extract from either Linum usitatissimum or Perilla frutescens (p<0.05) in a dose dependent manner. As for N-methyl-N'-nitro-N-nitrosoguamidine (MNNG)-induced mutagenicity, the methanol extracts (5 mg/assay) from Linum usitatissimum and Perilla frutescens showed 63% and 78% inhibitory rates, respectively, indicating that Perilla frutescens possessed stronger antimutagenic activity than did Linum usitatissimum. Inhibitory effects of methanol extracts from Linum usitatissimum and Perilla frutescens on the growth of human cancer cells (AGS, HT-29 and Hep 3B) appeared to increase dose dependently, and the inhibition was more effective against AGS and HT-29 compared to Hep 3B cells. Our results suggested that the methanol extract from Perilla frutescens showed stronger antimutagenic activity than that from Linum usitatissimumas assayed by the Ames mutagenic test, whereas the methanol extract from Linum usitatissimum was more effective than its counterpart for growth inhibition of human cancer cells. It is concluded that intake of Linum usitatissimum and Perilla frutescens as sources of omega-3 fatty acids will be beneficial for preventing cancer.

Intratumoral Administration of Dendritic Cells Combined with Hyperthermia Induces Both Local and Systemic Antitumor Effect in Murine Tumor Models (온열 요법 후 종양 내 주입한 수지상 세포의 국소 및 원격 항종양 효과)

  • Kwon Byung-Hyun;Kim Won-Taek;Kim Young-Kan;Kim Dong-Won
    • Radiation Oncology Journal
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    • v.24 no.1
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    • pp.51-57
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    • 2006
  • Puroose: We examined whether intratumoral (i.t.) administration of dendritic cells (DCs) into a treated tumor could induce local and systemic antitumor effects in a mouse tumor model. Methods and Materials: C57BL/6 mice were inoculated s.c. in the right and left thighs with MCA-102 fibrosarcoma cells on day 0 and on day 7, respectively. On day 7, the tumors (usually 6 mm in diameter) on the right thigh were heated by immersing the tumor-bearing leg in a circulating water bath at $43^{\circ}C$ for 30 min; thereafter, the immature DCs were i.t administered to the right thigh tumors. This immunization procedure was repeated on days 7, 14 and 21. The tumors in both the right and left thighs were measured every 7 days and the average sizes were determined by applying the following formula, tumor $size=0.5{\times}(length+width)$. Cytotoxicity assay was done to determine tumor-specific cytotoxic T-lymphocyte activity. Results: Hyperthermia induced apoptosis and heat shock proteins (HSPs) in tumor occurred maximally after 6 hr. For the local treated tumor, hyperthermia (HT) alone inhibited tumor growth compared with the untreated tumors (p<0.05), and furthermore, the i.t. administered DCs combined with hyperthermia (HT + DCs) additively inhibited tumor growth compared with HT alone (p<0.05). On the distant untreated tumor, HT alone significantly inhibited tumor growth (p<0.05), and also HT + DCs potently inhibited tumor growth (p<0.001); however, compared with HT alone, the difference was not statistically significant. In addition, HT + DCs induced strong cytotoxicity of the splenocytes against tumor cells compared to DCs or HT alone. Conclusion: HT + DCs induced apoptosis and increased the expression of HSPs, and so this induced a potent local and systemic antitumor response in tumor-bearing mice. This regimen may be beneficial for the treatment of human cancers.

Mycoplasma pneumoniae-induced production of proasthmatic mediators in airway epithelium (인체 기관지 상피세포에서 Mycoplasma pneumoniae 감염에 의한 천식 매개물질의 발현)

  • Kim, Kyung Won;Lee, Byung Chul;Lee, Kyung Eun;Kim, Eun Soo;Song, Tae Won;Park, Mi Yeoun;Sohn, Myung Hyun;Kim, Kyu-Earn
    • Clinical and Experimental Pediatrics
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    • v.49 no.9
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    • pp.977-982
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    • 2006
  • Purpose : There has been an increasing amount of literature concerning the association between Mycoplasma pneumoniae and asthma pathogenesis. Interleukin(IL)-6 stimulates the differentiation of monocytes, and can promote Th2 differentiation and simultaneously inhibit Th1 polarization. IL-8 is a potent chemoattractant and, it has been suggested, has a role in asthma pathogenesis. Nitric oxide (NO) synthesized by airway epithelium may be important in the regulation of airway inflammation and reactivity. Vascular endothelial growth factor(VEGF) has been reported to be a mediator of airway remodeling in asthma. We investigated the effects of M. pneumoniae on IL-6, IL-8, NO and VEGF production in human respiratory epithelial cells. Methods : A549 cells were cultured and inoculated with M. pneumoniae at a dose of 20 cfu/cell. After infection, the presence of M. pneumoniae in epithelial cell cultures was monitored by immunofluorescence and confirmed by polymerase chain reaction(PCR) detection. IL-6, IL-8 and VEGF were determined by an enzyme-linked immunosorbent assay and reverse transcriptase-polymerase chain reaction. NO was measured using the standard Griess reaction. Results : In A549 cells, M. pneumoniaeinduced IL-6, IL-8, NO and VEGF release in time-dependent manners. It also induced mRNA expression of IL-6, IL-8 and VEGF in similar manners. Conclusion : These observations suggest that M. pneumoniae might have a role in the pathogenesis of the allergic inflammation of bronchial asthma.

Hepatogenic Potential of Umbilical Cord Derived-Stem Cells and Human Amnion Derived-Stem Cells (사람의 제대 및 양막유래 줄기세포의 간세포로의 분화)

  • Kim, Ji-Young;Lee, Yoon-Jung;Park, Se-Ah;Kang, Hyun-Mi;Kim, Kyung-Sik;Cho, Dong-Jae;Kim, Hae-Kwon
    • Clinical and Experimental Reproductive Medicine
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    • v.35 no.4
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    • pp.247-265
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    • 2008
  • Objectives: Many types of liver diseases can damage regenerative potential of mature hepatocytes, hepatic progenitor cells or oval cells. In such cases, a stem cell-based therapy can be an alternative therapeutic option. We examined whether human amnion-derived mesenchymal stem cells (HAM) and human umbilical cord-derived stem cells (HUC) could differentiate into hepatocyte-like cells as therapeutic cells for the liver diseases. Methods: HAM and HUC were isolated from the amnion and umbilical cord of the volunteers after a caesarean section with informed consent. In order to differentiate these cells into hepatocyte-like cells, cells were cultivated in hepatogenic medium using culture plates coated with fibronectin. Effects of hepatocyte growth factor, L-ascorbic acid 2-phosphate, insulin premixture fibroblast growth gactor 4, dimethylsulfoxide, oncostatin M and/or dexamethasone were examined on the hepatic differentiation. After differentiation, the cells were analyzed by RT-PCR, immunocytochemistry, immunoblotting, albumin ELISA, urea assay and periodic acid-schiffs staining. Results: Initial fibroblast-like appearance of HAM and HUC changed to a round shape during culture in the hepatogenic medium. However, in all hepatogenic conditions examined, HUC secreted more amounts of albumin or urea into medium than HAM. Expression of some of hepatocyte-specific genes increased and expression of new genes were observed in HUC following cultivation in hepatogenic medium. Results of immunocytochemistry and immunoblotting analyses demonstrated that HUC secreted albumin into the culture medium. PAS staining further demonstrated that HUC could store glycogen inside of the cells. Conclusions: Both HUC and HAM could differentiate into albumin-secreting, hepatocyte-like cells. Under the same hepatogenic conditions examined, HUC more efficiently differentiated into hepatocyte-like cells compared with the HAM. The results suggest that HUC and HAM could be used as sources of stem cells for the cell-based therapeutics such as in liver diseases.

Antiangiogenic Effect of $As_4O_6$ on the Angiogenesis Induced by Vascular Endothelial Growth Factor (VEGF) in the Rat Cornea (랫드 각막에서 Vascular Endothelial Growth Factor(VEGF)로 유발시킨 신생혈관에 대한 $As_4O_6$의 혈관신생 억제효과)

  • Kwon Do-hyoung;Jang Jae young;Yi Na-young;Jeong Man-bok;Park Shin-ae;Kim Min-su;Nam Tchi-chou;Park Myung-jin;Bae Ill-ju;Rhee Chang-hun;Seo Kang-moon
    • Journal of Veterinary Clinics
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    • v.22 no.1
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    • pp.16-20
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    • 2005
  • The purpose of this study was to compare the antiangiogenic effects of As₄O/sub 6/ to those of As₂O₃ on the rat corneal micropocket model induced by VEGF. 20 ng VEGF impregnated pellets were used for angiogenic inducer on the rat cornea micropocket assay in this study. After ophthalmoscopic examination, Sprague-Dawley rats with normal cornea were implanted VEGF pellet. Total 60 eyes were used in this study. Control group only received VEGF pellet, As₂O₃ group followed oral administration of As₂O₃ at a dose of 50 mg/kg per day after VEGF pellet implantation and As₄O/sub 6/ group followed oral administration of As₄O/sub 6/ at a dose of 50 mg/kg per day after VEGF pellet implantation were classified. The eyes were examined under a surgical microscope daily on postoperative from day 3 to day 9 after pellet implantation. The number, length, clock hour of vascularization, and area of vessels in As₄O/sub 6/ group were significantly less evident than those of control group and As₂O₃ group (P < 0.05). In conclusion, As₄O/sub 6/ had better antiangiogenic effects on the new vessel induced by VEGF in the rat cornea.

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.