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Anticancer and Related Immunomodulatory Effects of Kwibi-tang on Non-small Cell Lung Carcinoma, NCI-H520, Xenograft Mice (귀비탕(歸脾湯)이 비소세포 폐암세포(NCI-H520) 이식 마우스에서 항암 및 면역 활성에 미치는 영향)

  • Son, Ji-Young;Choi, Hae-Yun;Kim, Jong-Dae
    • The Journal of Internal Korean Medicine
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    • v.33 no.4
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    • pp.387-404
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    • 2012
  • Objectives : This study was to observe anticancer and related immunomodulatory effects of Kwibi-tang extracts (KBTe) on non-small cell lung carcinoma (squamous epithelial carcinoma), NCI-H520, xenograft Balb/c nu-nu nude mice. Methods : Three different dosages of KBTe, 50, 100 and 200 mg/kg were orally administered once a day for 42 days from 11 days after tumor cell inoculation. Six groups, each of 8 mice per group were used in the present study. Changes in body weight, tumor volume and weight, lymphatic organs (spleen and popliteal lymph node), serum interferon (IFN)-${\gamma}$ levels, splenocytes NK cell activity and peritoneal macrophage activities, splenic tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-$1{\beta}$ and IL-10 contents were observed with tumor mass and lymphatic organ histopathology to detect anticancer and immunomodulatory effects. The results were compared with a potent cytotoxic anticancer agent, 5-FU (5-Fluorouracil) 30 mg/kg, intraperitoneal treatment (3-day intervals for 42 days, the optimal effective treatment regimes already confirmed). Results & Conclusions : This study suggest that over 50 mg/kg of KBTe showed favorable anticancer effects on the NCI-H520 cell xenograft with immunomodulatory effects. Although relatively lower anticancer effects were observed in KBTe 200 mg/kg treated mice as compared with 5-FU 30 mg/kg treated mice, no meaningful favorable immunomodulatory effects were observed after 5-FU treatment in the present study.

Prominent IL-12 Production and Tumor Reduction in Athymic Nude Mice after Toxoplasma gondii Lysate Antigen Treatment

  • Pyo, Kyoung-Ho;Jung, Bong-Kwang;Xin, Chun-Feng;Lee, You-Won;Chai, Jong-Yil;Shin, Eun-Hee
    • The Korean Journal of Parasitology
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    • v.52 no.6
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    • pp.605-612
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    • 2014
  • Toxoplasma gondii is an intracellular protozoan parasite that causes a Th1 cellular immunity. Our previous study showed that T. gondii lysate antigen (TLA) treatment in S180 tumor-bearing mice resulted in tumor reduction by suppressing CD31 expression, a marker of angiogenesis. In the present study, to investigate tumor suppressive effect of TLA under the absence of T lymphocytes, athymic nude mice were compared with euthymic mice in the anti-tumorigenic effect triggered by TLA in CT26 tumors. According to the results, intratumorally injected TLA reduced tumor growth and TIMP-1 level, a metastatic marker, in both euthymic and athymic mice. TLA treatment led to a sharp increase in IL-12 expression in serum cytokine profiling of athymic mice, and increased MyD88 signals in macrophages derived from the bone marrow, implying the activation of innate immunity. The selective induction of IL-12 by TLA treatment had an anti-tumorigenic effect.

Regeneration of Bovine Mammary Gland in Immunodeficient Mice by Transplantation of Bovine Mammary Epithelial Cells Mixed with Matrigel

  • Park, Hyun Jung;Lee, Won Young;Jeong, Ha Yeon;Song, Hyuk
    • International Journal of Stem Cells
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    • v.9 no.2
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    • pp.186-191
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    • 2016
  • Background and Objectives: With the global demand for dairy protein for consumption growing annually, there has been increasing activity in the research field of dairy protein synthesis and production. From a manipulation perspective, it is more difficult to use live cattle for laboratory studies on the production of milk as well as of dairy protein such as casein, as compared with using laboratory animals like rodents. Therefore, we aimed to develop a mouse model of bovine mammary alveolar ducts for laboratory-scale studies. We studied the formation of the bovine mammary gland ductal structure by transplanting the MAC-T bovine alveolar cell line into mice. Methods and Results: MAC-T cells ($1{\times}10^7$) were suspended in Matrigel and injected into the dorsal tissue of 8-week-old male BALB/C nude mice. Histological analysis of tissue dissected from the MAC-T cell-transplanted mice after 6 weeks showed the typical morphology of the tubuloalveolar female gland, as well as glands made up of branching ducts that were surrounded by smooth muscle with small alveoli budding off the ducts. In addition, the epithelial markers CK14 and CK18 were expressed within the duct-like structure. Prolactin was detected in the duct interior in these CK14+ and CK18+ cells but not in the non-transplanted MAC-T cells. Conclusions: These results showed that duct-like tissue had been successfully formed after 6 weeks of transplantation of the CK14+ and CK18+ MAC-T cells into mice dorsal tissue. This mouse model will be a useful tool for further research on the bovine mammary gland.

Anticancer Effects of Egg White Combined-Chalcanthite on NCI-H460 Tumor Regression Model (NCI-H460 폐암 유발 누드마우스 모형을 이용한 난담반의 항암 효과 연구)

  • Choi, Eun-A;Kim, Jung-Keun;Kim, Kyung-Soon;Choi, Jung-Eun;Cho, Chong-Kwan;Lee, Yeon-Weol;Yoo, Hwa-Seung
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.24 no.2
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    • pp.272-277
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    • 2010
  • This experimental study was performed to investigate the antitumor effect of Egg white combined-Chalcanthite (InSan 4, IS4) in xenografted nude mice with NCI-H460 human lung cancer cell. We cultured NCI-H460 cell lines and xenografted them on nude mice. These mice were divided into 3 groups; group with dose of 45 mg/kg IS4 orally, group with dose of 90 mg/kg IS4 orally, and the control group. They had been raised and treated for 28 days. We checked their body weight, tumor weight and volume twice a week, and their absolute organ weight, microhistological observation and biochemical blood analysis at the final day by sacrificing them. We also calculated their tumor inhibition rate (IR), mean survival time and percent increase in life span (% ILS). In this study, we observed that all of the IS4 treated mice have tumor regression, dosage-dependently, compared to the control group. Tumor weight and volume of high dose treated mice were smallest. IR increased in IS4 in a dose-dependent manner. Mean survival time and percent increase in life span (% ILS) in high-dose IS4 treatment group were the highest of the three groups. There was no significant difference in biochemical blood analysis, alanine phopsphatase (ALP), Calcium, creatinine (CRE), alanine transferase (ALT), and aspartate transaminase (AST) levels. The urea nitrogen (UN) level results significantly decreased by IS4 45 and 90 mg/kg (IS4 45 mg/kg, IS4 90 mg/kg, p<0.01). IS4 may have potential anti-tumor effect in a solid tumor induced by NCI-H460 without remarkable side effects.

Hiwi Knockdown Inhibits the Growth of Lung Cancer in Nude Mice

  • Liang, Dong;Dong, Min;Hu, Lin-Jie;Fang, Ze-Hui;Xu, Xia;Shi, En-Hui;Yang, Yi-Ju
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.2
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    • pp.1067-1072
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    • 2013
  • Hiwi, a human homologue of the Piwi family, plays an important role in stem cell self-renewal and is overexpressed in various human tumors. This study aimed to determine whether an RNA interference-based strategy to suppress Hiwi expression could inhibit tumor growth in a xenograft mouse model. A rare population of $SSC^{lo}\;Alde^{br}$ cells was isolated and identified as lung cancer stem cells in our previous study. Plasmids containing U6 promoter-driven shRNAs against Hiwi or control plasmids were successfully established. The xenograft tumor model was generated by subcutaneously inoculating with lung cancer stem cell $SSC^{lo}\;Alde^{br}$ cells. After the tumor size reached about 8 mm in diameter, shRNA plasmids were injected into the mice via the tail vein three times a week for two weeks, then xenograft tumor growth was assessed. In nude mice, intravenously delivery of Hiwi shRNA plasmids significantly inhibited tumor growth compared to treatment with control scrambled shRNA plasmids or the vehicle PBS. No mice died during the experiment and no adverse events were observed in mice administered the plasmids. Moreover, delivery of Hiwi shRNA plasmids resulted in a significant suppressed expression of Hiwi and ALDH-1 in xenograft tumor samples, based on immunohistochemical analysis. Thus, shRNA-mediated Hiwi gene silencing in lung cancer stem cells by an effective in vivo gene delivery strategy appeared to be an effective therapeutic approach for lung cancer, and may provide some useful clues for RNAi gene therapy in solid cancers.

Antitumor Effect of Hang-Am-Dan Non-boiled Water Extracts on NCI-H460 Tumor Regression Model

  • Kim, Jun-Lae;Kim, Kyung-Soon;Park, Jae-Woo;Lee, Yeon-Weol;Cho, Chong-Kwan;Yoo, Hwa-Seung
    • The Journal of Korean Medicine
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    • v.31 no.3
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    • pp.34-46
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    • 2010
  • Objective: This experimental study was performed to examine if Hang-Am-Dan non-boiled water extracts (HAD-N) induce apoptosis in human lung carcinoma NCI-H460 cells in vitro and inhibits the growth of NCI-H460 cell-transplanted solid tumor in vivo. Materials and Methods: We cultured NCI-H460 cell lines and xenografted them to nude mice. The mice were divided into 3 groups, NCI-H460 cell alone, NCI-H460 + 90 mg/kg HAD-N treated group, and NCI-H460 + 180 mg/kg HAD-N treated group, with seven mice per group. HAD-N was orally administrated every day for four weeks. We checked their body weight and tumor weight and volumes two times a week and their absolute organ weight and biochemical blood analysis at the final day by sacrificing them. We also calculated their tumor inhibition rate (IR), mean survival time and percent increase in life span (% ILS). Results: In this study, we observed that all of the HAD-N treated mice got smaller tumors. The more doses of HAD-N used, the less IR showed at the 8th day after starting this experiment. Tumor weight and volume of HAD-N treatment groups also decreased. Mean survival time and percent increase in life span (% ILS) in the high-dose HAD-N treatment groups were higher than those of other groups. The test substances in the blood level UN results showed reduction in the significance in both HAD-N 90 mg/kg and HAD-N 180 mg/kg (p<0.01). The blood level phosphatase results in HAD-N 90 mg/kg group compared to NCI-H460 cell alone group showed a reduction in significance (p<0.05). AST levels HAD-N 180 mg/kg group compared to NCI-H460 cell alone group significance as well (p<0.05). Conclusion: We suggest that the results of the in vivo study showed that HAD-N may have potential as a growth inhibitor of tumor-induced NCI-H460 of nude mice in spite of the shortcomings of this study. More studies to overcome those shortcomings and to find out significant antitumor mechanism will be needed.

Evaluation of the Various Artificial Skin Substitutes Implanted onto Nude Mice (누드마우스를 이용한 다양한 피부 대체물의 성능비교)

  • Lee, Won Jai;Lee, Dong Won;Hur, Jae Young;Lee, Young Dae;Park, Beyoung Yun;Rah, Dong Kyun
    • Archives of Plastic Surgery
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    • v.35 no.2
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    • pp.127-133
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    • 2008
  • Purpose: The purpose of this study is to evaluate the remodeling process of the various skin substitutes in 4th and 6th weeks following the transplantation when transplanted onto nude mice. Methods: Three types of artificial skin substitutes, such as PLGA scaffold with keratinocyte sheets(group 1), acellular human dermis($Surederm^{(TM)}$) and keratinocyte sheet(group 2), bioengineered skin($Neoderm^{(TM)}$)(group 3), were applied to the wound on nude mice. All mice were killed in 2, 4 weeks and/or 6 weeks after grafting and tissue samples were harvested from the back of mice. The changes in wound size, degree of angiogenesis, formation of basement membrane and epidermis, density of collagen fibers and neural restoration were examined. Results: There was no significant changes in wound size among the three groups. However, the size of wound decreased in the non-substituted group due to contracture. Degree of angiogenesis and systhesis of collagen or neurofilaments were mostly increased in bioengineered skin($Neoderm^{(TM)}$)(group 3), followed by acellular human dermis($Surederm^{(TM)}$) and keratinocyte sheet(group 2), PLGA scaffold with keratinocyte sheets (group 1). However, group 3 and group 2 showed similar thickness of basement membrane and epidermis. Conclusion: We found that degree of angiogenesis, formation of basement membrane and skin appendages, density of collagen fibers and neurofilaments can be the categories to evaluate the success of artificial skin substitution in early stages.

Adipose-Derived Stem Cell Coculturing Stimulates Integrin-Mediated Extracellular Matrix Adhesion of Melanocytes by Upregulating Growth Factors

  • Kim, Hyangmi;Yi, Nayoung;Do, Byung-Rok;Lee, Ai-Young
    • Biomolecules & Therapeutics
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    • v.27 no.2
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    • pp.185-192
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    • 2019
  • Coculture with adipose-derived stem cells (ADSCs) can stimulate proliferation and migration of melanocytes. To enhance outcomes of skin disorders caused by melanocyte loss or death, mixed transplantation with ADSCs has been suggested. However, role of cocultured ADSCs in proliferation and migration of melanocytes remains unclear. This study determined the effect of ADSCs on production of growth factors and expression levels of intergrins in primary culture of adult human melanocytes with or without ADSCs and in nude mice grafted with such melanocytes. Higher amounts of growth factors for melanocytes, such as bFGF and SCF were produced and released from ADSCs by coculturing with melanocytes. Relative levels of integrins ${\beta}1$, ${\alpha}5$, and ${\alpha}6$ as well as adhesion to fibronectin and laminin were increased in melanocytes cocultured with ADSCs. Such increases were inhibited by neutralization of bFGF or SCF. Relative levels of bFGF, SCF and integrins were increased in nude mice skin after grafting with melanocyte+ADSC cocultures. Collectively, these results indicate that ADSCs can stimulate proliferation and migration of melanocytes by increasing expression of integrins in melanocytes through upregulation of production/release of melanocyte growth factors such as bFGF and SCF.

Mode of Action of Coptidis Rhizoma Protein (CRP) and Its Activity Against Subcutaneous Candidiasis due to Candida albicans (황련단백질의 항캔디다 작용기전 및 항피부캔디다증 효과)

  • Lee, Jue-Hee;Shim, Jin Kie;Han, Yongmoon
    • YAKHAK HOEJI
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    • v.49 no.5
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    • pp.422-427
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    • 2005
  • Our previous data showed the protein isolated from Coptidis Rhizoma (CRP) had antifungal activity. In present study, we examined mode of action of the CRP and its activity against subcutaneous candidiasis due to C. albicans yeast cells. Results showed that the CRP blocked hyphal production from yeast form of C. albicans. The CRP also activated RAW 264.7 monocyte/macrophage cell line, which resulted in nitiric oxide (NO) production from the cells. This activation seemed to increase macrophage phagocytosis to destroy the invaders. Like other antimicrobial peptides, CRP was influenced by ionic strength, thus resulting in a decrease of antifungal activity. In murine model of a subcutaneous candidiasis, the sizes of infected areas of the nude mice given the CRP after subcutaneous injection of C. albicans yeast cells to the dorsal skin were $90\%$ less than those of the nude mice groups that received DPBS instead of the CRP. All data indicate that the CRP, which appeared to act like an antimicrobial peptide and to inhibit the morphological transition from blastoconidia, was effec­tive against the subcutaneous disease.

CELLULAR AND MOLECULAR CHARACTERIZATION OF ADENOID CYSTIC CARCINOMA OF THE SALIVARY GLANDS (침샘 선양낭성암종의 세포학적, 분자생물학적 특성에 관한 연구)

  • Park, Young-Wook
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.27 no.2
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    • pp.110-122
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    • 2005
  • Adenoid cystic carcinoma (ACC) of salivary glands has a protracted clinical course with perineural invasion, delayed onset of hematogenous metastasis, and poor responses to classical cytotoxic chemotherapic agents. Most deaths from salivary ACC are caused by lung metastases that are resistant to conventional therapy. Therefore, knowledge of cellular properties and tumor-host interactions that influence the dissemination of metastatic cells is important for the design of more effective therapy of salivary cancer. I determined in vitro expression of epidermal growth factor receptor (EGFR) and its downstream effectors and vascular endothelial growth factor receptor (VEGFR)-2 on a human salivary ACC cell line (ACC2). I also evaluated the expression of EGF and VEGF signaling molecules and metastasis-related proteins on human salivary ACC cells orthotopically growing in nude mice. In Western blot and immunohistochemical analyses, EGFR and VEGFR-2 were presented and phosphorylated in ACC2 cells. In human parotid cancer xenografts in nude mice, EGF and VEGF signaling molecules, IL-8, and MMP-9 were expressed at markedly higher levels than in normal parotid tissues. Moreover, tumor-associated endothelial cells of this orthotopic parotid tumor expressed phosphorylated VEGFR-2 and phosphorylated Akt, which is a cell-survival protein. These data show that those biomarkers can be molecular targets for therapy of salivary ACC, which has a propensity for delayed lung metastasis.