• Title/Summary/Keyword: CAM cell

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Effects of Caffeic Acid Methyl Ester from Mentha arvensis Linne var. piperascens on Function of E6 Oncoprotein of HPV 16 (박하에서 분리한 Caffeic Acid Methyl Ester가 자궁경부암 바이러스 발암단백질 E6의 기능에 미치는 영향)

  • Baek, Tae-Woong;Lee , Kyung-Ae;Ahn , Min-Jung;Joo, Hae-Hyung;Oh , Hyun-Cheol;Ahn , Jong-Seok;Cho , Yong-Kweon;Myung , Pyung-Keun;Yoon, Do-Young
    • YAKHAK HOEJI
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    • v.48 no.6
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    • pp.328-334
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    • 2004
  • Cervical cancer is one of the leading causes of female death. Viral oncoproteins E6 and E7 are selectively retained and expressed in carcinoma cells infected with HPV (Human pa pilloma virus) type 16 and cooperated in immotalization and transformation of primary keratinocyte. E6 and E7 oncoproteins interfere the functions of tumor suppressor proteins p53 and retinoblasoma protein (pRb), respectively. Among a lots of natural products, Mentha arvensis Linne var.piperascens have inhibitory effects on bindings between E6 oncoprotein and tumor suppressor p53, E3 ubiqutin- protein ligase (E6AP). HPV oncoprotein inhibitors from Mentha piperita L. were isolated by solvent partition and column chromatography (Silica gel, RP-18) and inhibitory compounds were finally purified by HPLC using an ELISA screening system based on binding between E6 and E6AP. The aim of this study is to identify the structure of inhibitory compounds and to investigate whether these compounds have inhibitory effects on functions of E6 oncoprotein. We investigated whether caffeic acid methyl ester (CAM) extracted from Mentha piperita L. could inhibit the function of E6 oncoprotein. CAM inhibited the in vitro binding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53 and also inhibited the proliferation of human cervical cancer cell lines (SiHa and CaSKi) in a dose response manner. These results suggest that CAM inhibited the function of E6 oncoprotein, suggesting that it can be used as a potential drug for the treatment of cervical cancers infected with HPV.

Relationship between ganglioside expression and anti-cancer effects of a plant-derived antibody in breast cancer cells

  • Ju, Won Seok;Song, Ilchan;Park, Se-Ra;Seo, Sang Young;Cho, Jin Hyoung;Min, Sung-Hun;Kim, Dae-Heon;Kim, Ji-Su;Kim, Sun-Uk;Park, Soon Ju;Ko, Kisung;Choo, Young-Kug
    • Journal of Plant Biotechnology
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    • v.46 no.3
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    • pp.217-227
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    • 2019
  • Production of therapeutic monoclonal antibodies (mAbs) using a plant platform has been considered an alternative to the mammalian cell-based production system. A plant-derived mAb CO17-1AK ($mAb^P$ COK) can specifically bind to various types of cancer cell lines. The target protein of $mAb^P$ COK is the epithelial cell adhesion molecule (EpCAM) highly expressed in human epithelial cancer cells, including breast and colorectal cancer cells. It has been hypothesized that its overexpression supports tumor growth and metastasis. A ganglioside is extended well beyond the surfaces of the various cell membranes and has roles in cell growth, inflammation, differentiation, and carcinogenesis. However, the regulation of EpCAM gene expression in breast cancers and the role of gangliosides in oncogenesis are unclear. Here, the purpose of this study was to determine the effects of $mAb^P$ COK on human breast cancer cell proliferation, apoptosis, and ganglioside expression patterns. Our results show that treatment with $mAb^P$ COK suppressed the growth of breast cancer cells and induced apoptotic cell death. It also upregulated the expression of metastasis-related gangliosides in breast cancer cells. Thus, treatment with $mAb^P$ COK may have chemo-preventive therapeutic effects against human breast cancer.

Expression and in vitro function of anti-cancer mAbs in transgenic Arabidopsis thaliana

  • Song, Ilchan;Kang, Yang Joo;Kim, Dae Heon;Kim, Mi Kyung;Ko, Kisung
    • BMB Reports
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    • v.53 no.4
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    • pp.229-233
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    • 2020
  • The anti-colorectal cancer monoclonal antibody CO17-1A (mAb CO), which recognizes the tumor-associated antigen EpCAM, was expressed in transgenic Arabidopsis plants. PCR and western blot analyses showed the insertion and expression of heavy chain (HC)/HC fused to the KDEL ER retention modif (HCK) and light chain (LC) of mAb CO and mAb CO with HCK (mAb COK) in Arabidopsis transformants. Both plant-derived mAbP CO and mAbP COK were purified from a biomass of approximately 1,000 seedlings grown in a greenhouse. In sandwich ELISA, both mAbP CO showed a slightly higher binding affinity for the target, EpCAM, compared to mAbM CO. In cell ELISA, both mAbsP COs showed binding affinity to the human colorectal cancer cell line SW480. Furthermore, mAbM CO, mAbP CO, and mAbP COK exhibited dose and timedependent regression effects on SW480 cells in vitro. In summation, both mAbP CO and mAbP COK, expressed in Arabidopsis, recognized the target antigen EpCAM and showed anti-proliferative activity against human colorectal cancer cells.

Study on driver's distraction research trend and deep learning based behavior recognition model

  • Han, Sangkon;Choi, Jung-In
    • Journal of the Korea Society of Computer and Information
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    • v.26 no.11
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    • pp.173-182
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    • 2021
  • In this paper, we analyzed driver's and passenger's motions that cause driver's distraction, and recognized 10 driver's behaviors related to mobile phones. First, distraction-inducing behaviors were classified into environments and factors, and related recent papers were analyzed. Based on the analyzed papers, 10 driver's behaviors related to cell phones, which are the main causes of distraction, were recognized. The experiment was conducted based on about 100,000 image data. Features were extracted through SURF and tested with three models (CNN, ResNet-101, and improved ResNet-101). The improved ResNet-101 model reduced training and validation errors by 8.2 times and 44.6 times compared to CNN, and the average precision and f1-score were maintained at a high level of 0.98. In addition, using CAM (class activation maps), it was reviewed whether the deep learning model used the cell phone object and location as the decisive cause when judging the driver's distraction behavior.

Identification of a Technique Optimized for the Isolation of Spermatogonial Stem Cells from Mouse Testes

  • Han, Na Rae;Park, Hye Jin;Lee, Hyun;Yun, Jung Im;Choi, Kimyung;Lee, Eunsong;Lee, Seung Tae
    • Journal of Embryo Transfer
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    • v.33 no.4
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    • pp.327-336
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    • 2018
  • To date, there are no protocols optimized to the effective separation of spermatogonial stem cells (SSCs) from testicular cells derived from mouse testes, thus hindering studies based on mouse SSCs. In this study, we aimed to determine the most efficient purification method for the isolation of SSCs from mouse testes among previously described techniques. Isolation of SSCs from testicular cells derived from mouse testes was conducted using four different techniques: differential plating (DP), magnetic-activated cell sorting (MACS) post-DP, MACS, and positive and negative selection double MACS. DP was performed for 1, 2, 4, 8, or 16 h, and MACS was performed using EpCAM ($MACS^{EpCAM}$), Thy1 ($MACS^{Thy1}$), or GFR ${\alpha}1$ ($MACS^{GFR{\alpha}1}$) antibodies. The purification efficiency of each method was analyzed by measuring the percentage of cells that stained positively for alkaline phosphatase. DP for 8 h, $MACS^{Thy1}$ post-DP for 8 h, $MACS^{GFR{\alpha}1}$, positive selection double $MACS^{GFR{\alpha}1/EpCAM}$, and negative selection double $MACS^{GFR{\alpha}1/{\alpha}-SMA}$ were identified as the optimal protocols for isolation of SSCs from mouse testicular cells. Comparison of the purification efficiencies of the optimized isolation protocols showed that, numerically, the highest purification efficiency was obtained using $MACS^{GFR{\alpha}1}$. Overall, our results indicate that $MACS^{GFR{\alpha}1}$ is an appropriate purification technique for the isolation of SSCs from mouse testicular cells.

Biotransformation of Flavone by CYP105P2 from Streptomyces peucetius

  • Niraula, Narayan Prasad;Bhattarai, Saurabh;Lee, Na-Rae;Sohng, Jae Kyung;Oh, Tae-Jin
    • Journal of Microbiology and Biotechnology
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    • v.22 no.8
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    • pp.1059-1065
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    • 2012
  • Biocatalytic transfer of oxygen in isolated cytochrome P450 or whole microbial cells is an elegant and efficient way to achieve selective hydroxylation. Cytochrome P450 CYP105P2 was isolated from Streptomyces peucetius that showed a high degree of amino acid identity with hydroxylases. Previously performed homology modeling, and subsequent docking of the model with flavone, displayed a reasonable docked structure. Therefore, in this study, in a pursuit to hydroxylate the flavone ring, CYP105P2 was co-expressed in a two-vector system with putidaredoxin reductase (camA) and putidaredoxin (camB) from Pseudomonas putida for efficient electron transport. HPLC analysis of the isolated product, together with LC-MS analysis, showed a monohydroxylated flavone, which was further established by subsequent ESI/MS-MS. A successful 10.35% yield was achieved with the whole-cell bioconversion reaction in Escherichia coli. We verified that CYP105P2 is a potential bacterial hydroxylase.

Study on Antitumor Activity of Sobokchukeotang and Kamisobokchukeotang (소복축어탕과 가미소복축어탕이 항암활성에 미치는 영향)

  • 신원웅;최주선;길재호;김성훈
    • The Journal of Korean Medicine
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    • v.22 no.2
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    • pp.22-30
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    • 2001
  • This study was attempted to investigate the anti-tumor and anti-metastatic effects of Sobokchukeotang(SBCT) and Kamisobokchukeotang(KSBCT). Cytotoxicity against various cancer cell lines, anti-adhesion, pulmonary colonization, anti-angiogenesis, and T/C% were evaluated. SBCT and KSBCT exhibited no cytotoxicity against HT-1080, A549, SK-OV-3, B16-F10 and SK-Mel-2 cell lines. In inhibitory effect on DNA topoisomerase I, the $IC_{50S}$ were shown $250-500{\;}\mu\textrm{g}/ml$ of SBCT and $62.5-125{\;}\mu\textrm{g}/ml$ of KSBCT respectively. In the in vivo experiments, SBCT(135.98%) and KSBCT(151.92%) apparently increased the life span of mice bearing sarcoma-180. KSBCT significantly inhibited the adhesion of HT-1080 to complex extracellular matrix in a dose-dependent manner in contrast to SBCT. In pulmonary colonization assay by B16-F10, a number of colonies in the lungs were decreased more significantly in KSBCT group than those in SBCT group. In vitro neovascularization and CAM assay, angiogenesis was more significantly inhibited in KSBCT-treated group than in SBCT- treated group. Above results suggests that KSBCT is more effectively applied to prevention and treatment of cancer than SBCT.

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Study on Antitumor Activity of Kamibojungikgi-tang (가미보중익기탕이 항암활성 및 항전이효과에 관한 연구)

  • Lee Byung Ju;Kim Dong Hee;Lee Hyo Jeong;Kim Sung Hoon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.3
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    • pp.765-770
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    • 2003
  • To explore the possible cancer agent from oriental prescriptions, we have examined its antitumor and anti metastatic activities of Kamibojungikgi-tang(KBIT). KBIT extracts exhibited cytotoxicity against P388, A549 and B16-F10 cell lines in a dose-dependent manner and showed antiadhesive effect of A549 cell to complex extracellular matrix at 1 ㎎/㎖ in vitro. In DNA topoisomerase I assay, KBIT extracts showed strong inhibitoty effect in a dose-dependent manner. In pulmonary colonization assay with B16BL6, a number of colonies in the lungs were decreased effectively in KBIT treated group as compared with control group. Moreover, in CAM assay, KBIT extracts significantly inhibited angiogenesis at 15㎍/egg as compared with control. The T/C% was 141% in KBIT treated group in S-180 bearing ICR mice. From the above results it was concluded that KBIT had antitumor and anti metastatic activities. So it is expected to be clinically helpful on the prevention and treatment of cancer, although it is still necessary to study its mechanism on molecular biology and immunology.

Study on Antitumor and Antimetastatic Effect of Kamigedang-tang (가미저당탕의 항암 및 항전이 효과에 관한 연구)

  • Lee Dong Hoon;Kim Dong Hee;Kang In Cheol;Park Young Mi;Song Gyu Yong;Kim Sung Hoon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.3
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    • pp.472-478
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    • 2002
  • The purpose of this study was to investigate the effect of Kamigedang-tang(KGDT) water extract on the antitumor and anti metastatic activity. The results were summarized as follows: KGDT extracts exhibited a significant cytotoxicity against P388, SK-MEL-2, SK-OV-3, and B16-F10 cell lines and showed significant inhibitoty effect on DNA topoisomerase I from calf thymus. The T/C% was 122.9% in KGDT treated group in S-180 bearing ICR mice. Also, KGDT extracts exhibited efficient affect adhesive effect of A549 cell to complex extracellular matrix. In CAM assay, KGDT extracts inhibited angiogenesis at 15㎍/egg concentration insignificantly as compared with control. These results suggested that KGDT extracts might be usefully applied for prevention and treatement of cancer.

Study on Antitumor Activity of Samiyeongeontanggamibang(SYTG) (사미연견탕가미방(四味軟堅湯加味方)이 항암(抗癌) 및 항전이(抗轉移) 효과(效果)에 미치는 영향)

  • Bae, Moon-Yong;Kang, In-Cheol;Kim, Sung-Hoon
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.5 no.1
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    • pp.33-46
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    • 1999
  • To evaluate the antitumor and antimetastatic effects of Samiyeongeontanggamibang(SYTG), We have examined whether SYTG can inhibit the growth of several tumor cell lines, tumor cell adhesion, experimental tumor metastasis and increase survival rate of tumor-bearing mice by inhibition of DNA topoisomrase activity. The results were obtained as follows: 1. SYTG extracts revealed an efficient cytotoxicity against A549, SK-OV-3, B16-F10, and SK-MEL-2 cell lines. 2. SYTG extracts inhibited DNA topo-isomerase I from calf thymus. 3. The T/C% in S-180 bearing ICR mice treated with SYTG was 115.8% 4. SYTG extracts significantly inhibited adhesion of A549 cell to complex extracellular matrix. 5. In pulmonary colonization assay, SYTG suppressed lung metastases in tumor cell-injected mice. 6. In CAM assay, SYTG extracts inhibited angiogenesis at $15{\mu}g/egg$ concentration as compared with control. These results suggested that SYTG extracts might be a potent inhibitor of cancer.

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