• Title/Summary/Keyword: in vitro cancer research

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Inhibitory Effects of 23 Korean Local Plant Extracts on Aldo-keto reductase family 1 B10 (AKR 1 B10) (한국 자생식물 추출물 23종의 Aldo-keto reductase family 1 B10 (AKR 1 B10) 효소 억제효과)

  • Lee, Joo-Young;Song, Dae-Geun;Jung, Sang-Hoon;Kim, Jong-Hwan;Ahn, Soo-Young;Nho, Chu-Won;Pan, Cheol-Ho
    • Korean Journal of Pharmacognosy
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    • v.40 no.3
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    • pp.238-243
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    • 2009
  • We examined ethanol extracts prepared from 23 Korean local plants obtained in Pyeongchang, Gangwon-do for their inhibitory effects on recombinant human AKR 1 B10 (rhAKR1B10) in vitro. To do this, rhAKR1B10 was first expressed in E. coli as a biological active form and purified by using Ni-affinity chromatography followed by gel permeation chromatography. Then, rhAKR1B10 was used for screening out 23 Korean local plant extracts having an inhibitory activity against itself. Among them, six extracts showed more than 50% inhibition of rhAKR1B10 activity at the concentration of $10{\mu}g$/ml. Especially, the extracts of Ligularia fischeri var. spiciformis Nakai and Rhus trichocarpa Miq. were the most potent because their $IC_{50}$ values were 2.94 and $2.00{\mu}g$/ml, respectively.

Replicative Senescence in Cellular Aging and Oxidative Stress (세포 노화에 있어서 복제 세네센스 현상과 산화적 스트레스의 영향)

  • 박영철
    • Toxicological Research
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    • v.19 no.3
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    • pp.161-172
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    • 2003
  • Explanted mammalian cells perform a limited number of cell division in vitro and than are arrested in a state known as replicative senescence. Such cells are irreversibly blocked, mostly in the G1 phase of cell cycle, and are no longer sensitive to growth factor stimulation. Thus replicative senescence is defined as a permanent and irreversible loss of replicative potential of cells. For this characteristic, replicative senescence seems to evolve to protect mammalian organism from cancer. However, senescence also contributes to aging. It seems to decrease with age of the cell donor and, as a form of cell senescence, is thought to underlie the aging process. Extensive evidence supports the idea that progressive telomere loss contributes to the phenomenon of cell senescence. Telomeres are repetitive structures of the sequence (TTAGGG)n at the ends of linear chromosomes. It has been shown that the average length of telomere repeats in human somatic cells decreases by 30∼200 bp with each cell division. It is generally believed that when telomeres reach a critical length, a signal is activated to initiate the senescent program. This has given rise to the hypothesis that telomeres act as mitotic clocks to regulate lifespan. One proposes that cumulative oxidative stress, mainly reactive oxygen species generated from mitochondria, may mainly cause telomere shortening, accelerating aging. Here, the biological importance and mechanism of replicative senescence were briefly reviewed. Also it was summarized that how oxidative stress affects replicative senescence and telomere shortening.

Development and Application of Artificial Skin Using Tissue Engineering (조직배양공학을 이용한 인공피부의 개발 및 응용)

  • Yang, Eun-Kyung;Park, Sue-Nie;Park, Jung-Keug
    • Proceedings of the KOSOMBE Conference
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    • v.1995 no.11
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    • pp.14-17
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    • 1995
  • An in vitro construct of three dimensional artificial skin equivalent has been engineered using human cervical epithelial cells and human foreskin fibroblasts with a matrix of bovine type I collagen. Two cell lines were established from cervical uteri cancer tissues which have the HPV(human papillomavirus)18 genome. These two cell lines came from the same origin but have slight differencies in growth rate and tumorigenicity. The organotypic raft culturing of epithelial cells were accomplished at air-liquid interface. The differentiation related characteristics were examined by immunohistochemistry using monoclonal antibodies against EGFreceptor, cytokeratin 5/6/18 as proliferation markers and against filaggrin, involucrin, and cytokeratin 10/13 as differentiation marker. We have obtained the stratification and the differentiation in the artificial skin equivalent, and differentiation-related proteins were expressed more in the C3-artificial skin, and proteins of proliferation were expressed more in the C3N-artificial skin, relatively. We found that reconstituted artificial skin have the same characteristics of differentiation proteins of original tissue or cells of human body.

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Effect of Sojagangqi-tang on the Immunomodulatory Action (소자강기탕의 면역조절효과)

  • Shin Hyun Jong;Jeong Sang Hun;Jeong Han Sol;Lee Kwang Gyu
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.5
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    • pp.1182-1187
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    • 2003
  • The purpose of this research was to investigate the effect of Sojagangqi-tang(SJGQT) on the immune cell activity. The addition of SJGQT enhanced the proliferation of cultured-mice splenocytes and thymocytes. Administration of SJGQT(250 mg/kg) accelerated the subpopulation of splenic T lymphocytes especially CD/sup 4+/-TH cells in BALB/c mice. But high concentration(500 mg/kg) of SJGQT decreased the splenic T, B lymphocytes and thymic Tc (CD/sup 8+/) lymphocytes. Oral administration of SJGQT(250 mg/kg) significantly enhanced the production of IFN-γ and IL-4 in mice serum. And also, the addition of SJGQT(100 ㎍/ml) inhibited the proliferation of cultured-Jurkat leukemia cells in vitro. These results suggest that SJGQT have a cellular immuno-modulatory effect and anti-cancer property action

Immunomodulatory activity of phytoprotein isol Acanthopanax senticosus : Regulation of CTL responses and activation of macrop

  • Lee Seok Won;Lee Soo Jin;Park Jeon Ran;Kim Ha na;Ahn Kyoo Seok;Kim Jung-Hyo;Baek Nam-in;Kim Sung Hoon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.1
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    • pp.230-235
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    • 2004
  • We previously reported that the phytoprotein extracted from Acanthopanax senticosus (PA) had anti-carcinogenic anti-metastatic activity via increase of inhibition of gap junctional intercellular communication. In the present study investigated the immunomodulatory mechanism of phytoprotein isolated from the stem bark of Acanthopanax sentic (PA). PA was found to significantly stimulate macrophages producing TNF-α and IL-1β in vitro. It also showed tumori activity indicating that PA had the ability to stimulate macrophage directly. Moreover, PA induced the CDB/sup +/ CTL cy responses to recognize antigen on the B16-BL6 melanoma cells. Treatment of PA with B16-BL6 melanoma cells increased the proliferation of splenocytes compared with untreated control. These results demonstrate that PA immunomodulatory activity suggesting a useful anti-tumor agent applicable to treatment and prevention of cancer.

Synthesis and Antitumor Activity of Phthalimide-Based Polymers Containing Camptothecin

  • Lee, Neung-Ju
    • Macromolecular Research
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    • v.11 no.1
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    • pp.47-52
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    • 2003
  • The objective of this study was to develop a polymeric drug delivery system for camptothecin (CPT), capable of improving its therapeutic index and reducing its side effects. A monomeric conjugate, 3,6-endo-methylene-1,2,3,6-tetrahydrophthalimidoethanoylcamptothecin in (ETECPT) between CPT and 3,6-endo-methylene-1,2,3,6-tetrahydrophthalimidoethanoic acid was synthesized. Its homo-and copolymer with acrylic acid (AA) were prepared by photopolymerization using 2,2-dimethoxy-2-phenylacetophenone (DMP) as a photoinitiator. The monomer and its polymers were characterized by IR, $^1$H- and $^{13}$ C-NMR spectra. The ETECPT content in poly(ETECPT-co-AA) obtained by elemental analysis was 82 wt%. The number-average molecular weights of the polymers determined by gel permeation chromatography were as follows: M$_{n}$ = 11,400 for poly(ETECPT), M$_{n}$ = 17,900 for poly(ETECPT-co-AA). The $IC_{50}$/ values of ETECPT and its polymers against cancer cells were much larger than that of CPT. Our results from the in vivo antitumor activity indicated that all polymers show high antitumor activity than CPT at a dose of 100 mg/kg./kg.

Antiestrogenic Effects of Marijuana Smoke Condensate and Cannabinoid Compounds

  • Lee Soo Yeun;Oh Seung Min;Lee Sang Ki;Chung Kyu Hyuck
    • Archives of Pharmacal Research
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    • v.28 no.12
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    • pp.1365-1375
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    • 2005
  • The antiestrogenic effects of marijuana smoke condensate (MSC) and three major cannabinoids, i.e., $\bigtriangleup^{9}$-tetrahydrocannabinol (THC), cannabidiol (CBD), and cannabinol (CBN), were evaluated using in vitro bioassays, viz., the human breast cancer cell proliferation assay, the recombinant human estrogen receptor (ER) competitive binding assay, and the reporter gene assay. The inhibitory effects on estrogen were also examined using the ethoxyresorufin-O­deethylase (EROD) assay, the aromatase assay, and the 17$\beta$-estradiol ($E_{2}$) metabolism assay. The results showed that MSC induced the antiestrogenic effect via the ER-mediated pathway, while THC, CBD, and CBN did not have any antiestrogenic activity. This suggests that the combined effects of the marijuana smoke components are responsible for the antiestrogenicity of marijuana use. In addition, MSC induced the CYP1A activity and the $E_{2}$ metabolism, but inhibited the aromatase activity, suggesting that the antiestrogenic activity of MSC is also related to the indirect ER-dependent pathway, as a result of the depletion of the in situ $E_{2}$ level available to bind to the ER. In conclusion, pyrogenic products including polycyclic aromatic hydrocarbons (PAHs) in the non-polar fraction, which is the most biologically active fraction among the seven fractions of MSC, might be responsible for the antiestrogenic effect.

Establishment and characterization of gastric surface mucous cell lines (GSM06 and GSM10) from transgenic mice harboring temperature-sensitive simian virus 40 large T-antigen gene

  • Tabuchi, Yoshiaki;Sugiyama, Norifumi;Horiuchi, Tadashi;Furuhama, Kazuhisa;Obinata, Masuo;Furusawa, Mitsuru
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.04a
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    • pp.131-136
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    • 1994
  • In the present study, in order to make an in vitro model of gastric mucosa for physiological and pharmacological studies, we established two immortalized gastric surface mucous cell lines (GSM06 and GSM10), which produce periodic acid-Schiff (PAS)-and concanavalin A (Con A)-positive glycoproteins, from a primary culture of gastric fundic mucosal cells of adult transgenic mice harboring a temperature-sensitive simian virus 40 large T-antigen gene 〔1]. Gastric fundic mucosal cells were isolated as a modification of a previously described method for rats by Schepp et al. (2). The isolated gastric fundic mucosal cells were cultured in DME/F12 medium supplemented with 2% fetal bovine serum (FBS), 1% ITES (consisting of 2 mg/1 insulin, 2 mgg/1 transferrin, 0.122 mg/1 ethanolamine and 0.00914 mg/1 sodium selenite) and 10 ng/ml recombinant epidermal growth factor (EGF) in a collagen-coated culture dish. To remove fibroblastic cells from the culture, gastric mucosal cells were incubated in the culture medium containing dispase (25 U/ml) for 24 h. The cells, uncontaminated with fibroblastic cells, were then cloned by colony formation. In our series of three attempts, two cell lines (GSM06 and GSM10) have been established at last. The cells proliferated, attached to the dish ana grew until confluent monolayers were formed, and maintained tight contact with neighboring cells. Both GSM06 and GSM10 cells have now been in culture for more than 9 months with regular passaging. The either cell produced

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Evaluating the effect of conditioned medium from mesenchymal stem cells on differentiation of rat spermatogonial stem cells

  • Hoda Fazaeli;Mohsen Sheykhhasan;Naser Kalhor;Faezeh Davoodi Asl;Mojdeh Hosseinpoor Kashani;Azar Sheikholeslami
    • Anatomy and Cell Biology
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    • v.56 no.4
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    • pp.508-517
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    • 2023
  • In cancer patients, chemo/radio therapy may cause infertility by damaging the spermatogenesis affecting the self-renewal and differentiation of spermatogonial stem cells (SSCs). In vitro differentiation of stem cells especially mesenchymal stem cells (MSCs) into germ cells has recently been proposed as a new strategy for infertility treatment. The aim of this study was to evaluate the proliferation and differentiation of SSCs using their co-culture with Sertoli cells and conditioned medium (CM) from adipose tissue-derived MSCs (AD-MSCs). Testicular tissues were separated from 2-7 days old neonate Wistar Rats and after mechanical and enzymatic digestion, the SSCs and Sertoli cells were isolated and cultured in Dulbecco's modified eagle medium with 10% fetal bovine serum, 1X antibiotic, basic fibroblast growth factor, and glial cell line-derived neurotrophic factor. The cells were treated with the CM from AD-MSCs for 12 days and then the expression level of differentiation-related genes were measured. Also, the expression level of two major spermatogenic markers of DAZL and DDX4 was calculated. Scp3, Dazl, and Prm1 were significantly increased after treatment compared to the control group, whereas no significant difference was observed in Stra8 expression. The immunocytochemistry images showed that DAZL and DDX4 were positive in experimental group comparing with control. Also, western blotting revealed that both DAZL and DDX4 had higher expression in the treated group than the control group, however, no significant difference was observed. In this study, we concluded that the CM obtained from AD-MSCs can be considered as a suitable biological material to induce the differentiation in SSCs.

Radioprotective effect of naringin and naringenin against cellular damage and oxidative stress of γ-irradiated mice (감마선을 조사한 마우스의 세포 손상과 산화적 스트레스에 대한 나린진과 나린제닌의 방사선방호 효과)

  • Kang, Jung Ae;Kim, Hye Rim;Yoon, Seon Hye;Jang, Beom-Su;Choi, Dae Seong;Park, Sang Hyun
    • Korean Journal of Food Science and Technology
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    • v.49 no.6
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    • pp.659-667
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    • 2017
  • The present study was designed to evaluate the antioxidant activity and radioprotective effects of Naringin and Naringenin in ${\gamma}$-irradiated mice. The antioxidant activity of Naringin and Naringenin was evaluated by 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and ferric reducing antioxidant power (FRAP) assays. Healthy female BALB/c mice were administered Naringin and Naringenin orally ($90{\mu}M/dose$ and $180{\mu}M/dose$) for 7 consecutive days prior to ${\gamma}$-irradiation (6 Gy). Naringenin displayed a much higher antioxidant activity in ABTS and FRAP than naringin. ${\gamma}$-irradiation resulted in cellular damage with decreased spleen and thymus indices and white blood cells (WBC) count. Additionally, ${\gamma}$-irradiation significantly increased lipid peroxidation and decreased the levels of antioxidant enzymes and glutathione (GSH) in the liver tissue. Strikingly, prior administration of Naringenin resulted in considerable prevention of these symptoms. Protection against ${\gamma}$-irradiation-induced cellular damage by Naringenin is likely due to its higher its antioxidant activity. Together, these results confirm that Naringenin is a potent antioxidant and radioprotector.