Anti-angiogenesis and anti-cell adhesion effects were investigated with different dose of Camellia japonica leaf (CJL) extract for applying anti-cancer, anti-metastasis and anti-obesity. Cytotoxicity on HUVECs was very low at 200 ug/mL of CJL-extract. Anti-angiogenic ratio at increasing dose of 1.5 ug/mL, 3.0 ug/mL, 15 ug/mL and 30 ug/mL was showed 30.7%, 38.5%, 53.8%, and 70.0%, respectively. Also, anti-cell adhesion effect at concentration of $50{\mu}g{/well},\;100{\mu}g{/well}\;and\;200{\mu}g{/well}$ was expressed on E-selectin by 46.7%, 66.7%, and 86.76%, on VCAM-1, 23.0%, 61.5%, and 84.6%, and on ICAM-1, 11%, 55.5%, and 88.8%, respectively. For inquiring anti-angiogenesis mechanism, when western blot was performed with different dose of CJL extract, signal molecules of VEGFR-2, $\beta$-catenin and PI3-K were suppressed. As the signal transduction from VEGFR-2, $\beta$-catenin and PI3-K to NF-${\kappa}$B was interupted, angiogenesis could not be occurred causing not activated NF-kB. C. japonica leaf (CJL) is a useful herb for developing therapeutics of angiogenesis related diseases such as cancer, metastasis, rheumathioid arthritis and obesity.
Park, Mi Na;Jang, Hyun-Jun;Keum, Dae Ho;Choi, Jin Ae;Yoo, Jae Gyu;Byun, Sung June;Park, Jong Ju;Ji, Ju Young;Lee, Kyung-Tai;Kim, Tae-Hun;Lee, Hyun-Jeong
Korean Journal of Poultry Science
/
v.40
no.4
/
pp.299-304
/
2013
Tracheal epithelial cells (TECs) are an important tool for studies of viral respiratory diseases. Primary TECs have been cultured from human, mouse and hamster. It is also necessary to diagnose viral respiratory disease and reveal infection mechanisms in chicken. In this study, we isolated tracheal epithelial layers from tracheal of 20-day-old chicks and cultured primary TECs from the isolated layers. Ciliated cells which were a typical morphology of TECs were observed in cultured primary TECs and maintained until cell passage 5 (15 to 20 days). When we analyzed expression patterns of epithelial marker genes (retinoic acid responder, FGF-binding protein, virus activating protease (VAP) in TECs compared to immortalized chicken embryonic fibroblast cell line (DF-1), all the marker genes are highly expressed in TECs than in DF-1. When TECs were cultured with 0.1 and 1 MOI of ND virus (rNDV-GFP strain) to test the susceptibility of TECs for ND virus, 12.6% and 48.2% of the incubated TECs were infected respectively. In addition, when DF-1 was incubated with 1 MOI of ND virus, the virus infection rate of DF-1 was three times lower than the virus infection rate of TECs. These data could contribute to study infection mechanisms of viral respiratory diseases and control them in chicken.
Background : The role of second-line chemotherapy in the treatment of advanced non-small cell lung cancer (NSCLC) is known to be limited. Recently, ZD1839, the small molecule epidermal growth factor receptor-tyrosine kinase inhibitor, has been developed and has shown anti-tumor activity in patients with solid malignant tumors including lung cancer. We evaluated the response rate and toxicities of ZD1839 in patients with advanced NSCLC which has progressed after previous chemotherapy. Patients and Methods : We examined 83 patients with advanced NSCLC treated with ZD1839 for more than 1 month in Korea Cancer Center Hospital during the period from January 2002 to September 2003. All the patients were enrolled in the international expanded access program (EAP) with ZD1839 by AstraZeneca. The administered dose of ZD1839 was 250 mg once daily. Chest radiography and laboratory tests were followed-up. We evaluated the response rate, median survival, and toxicity after treatment. Results : Median age of the patients was 59 years (range 33-76). The most predominant cell type was adenocarcinoma and the most stage of the patients was IV. ECOG performance status was as follows; grade 0-1 in 10, grade 2 in 42, and grade 3 in 31 patients. Partial response was achieved in 12 patients (14.5%). Median overall survival was 9.2 (range 1.3-21.6+) months and median time to progression was 3.1 (range 1-21.2+) months. The most common adverse effect of ZD1839 was skin eruption which developed in 25 patients (25.8%). Significantly higher response rate and survival was found in patients with adenocarcinoma or good performance status. Conclusion : ZD1839 showed modest activity and tolerable toxicity in the treatment for patients with NSCLC which has progressed after previous chemotherapy.
Kim, Gwang-Seok;Sung, Jae-Hyun;Choi, Je-Yong;Ryou, Hyun-Mo
The korean journal of orthodontics
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v.23
no.2
s.41
/
pp.229-248
/
1993
[ $Interferon-\gamma$ ] has been suggested as a cytokine of connective tissue stabilizer. In addition, it has also been demonstrated that this cytokine inhibited bone remodeling activities of the bone derived cells. In order to illuminate the effects of this cytokine in orthodontic force induced bone remodeling, it was administered to primary cultured periodontal ligament cells which have been known to have some osteoblast like characteristics. $Interferon-\gamma$ slightly decreased $[^3H]thymidine$ incorporation rate without a significant change in the total cellular DNA content up to 1000 U/ml, which meant these doses were not cytotoxic to the cell. Total protein synthesis was not influenced by various concentration of interferon-y whether it was determined by the $[^3H]proline$ incorporation rate or by the Lowry smethod. The effect of $interferon-\gamma$ on the individual protein was, however, differential, ie, it increased $[^3H]proline$ incorporation into the noncollagenous protein marginally, while it decreased $[^3H]proline$ incorporation into the collagen, so that it caused dose-dependent suppression of the relative collagen synthesis. On the contrary, the fibronectin synthesis determined by the ELISA was increased by 1000 U/ml of $interferon-\gamma$. The differential effects of the interferon-y on the collagen and fibronectin synthesis exhibited not only their protein level but also the steady state mRNA level. $Interferon-\gamma$ decreased steady state level of ${\alpha}1(I)$ procollagen mRNA significantly, while showing no significant changes in the fibronectin mRNA level. In addition to this, it was also found that indomethacin did not affect on the $interferon-\gamma$ induced collagen decrease in this cell, which meant prostaglandins were not involed in the process of $interferon-\gamma$ induced collagen decrease. So it can be concluded that the incubation of periodontal ligament cells with 1000 U/ml of $interferon-\gamma$ for 24 hr showed differential effects on the type I collagen and fibronectin gene expression. The decrease in relative collagen synthesis in the protein level was related with decrease in the steady state level of mRNA, while the increase in the fibronectin synthesis in the protein level was not correlated with the mRNA level.
α-Asarone is the main component of Acorus gramineus, which is a widely used oriental traditional medicine. A. gramineus is known to have a variety of medicinal effects, such as anti-gastric ulcer, antiallergy and antioxidant activity. It is also known to inhibit the release of histamine. However, the mechanism of its action remains unclear in humans. In this study, the effects of α-asarone on matrix metalloproteinase (MMP) and its antioxidant effect in a cell-free system were examined in HT1080 cells. In an MTT assay, the effect of α-asarone on cell viability showed no cytotoxicity below 16 μM. In an antioxidant assay, α-asarone increased reducing power in a dose-dependent manner but not the scavenging activity of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals. In addition, α-asarone exhibited the protective effect against DNA oxidation induced by hydroxyl radicals produced by the Fenton reaction. Furthermore, in a gelatin disk assay, α-asarone enhanced collagenase activity. It also increased the activities of MMP-2 and MMP-9 stimulated by phorbol 12-myristate 13-acetate (PMA) in a gelatin zymography. On the other hand, the activity of MMP-9 stimulated by phenazine methosulfate (PMS) but not that of MMP-2 was increased in the presence of α-asarone. These findings suggest that α-asarone could be a candidate for the prevention and treatment of pathological diseases related to oxidative stress and MMPs.
In the present investigation, we studied the modulating effects of caffeic acid, chlorogenic acid, and (-)-epigallocatechin-3-gallate(EGCG) on the methylation status of promoter regions of cell cycle regulator, p16, in human breast cancer T-47D cells. We demonstrated that treatment of T-47D cells with caffeic acid, chlorogenic acid, or EGCG partially inhibited the methylation status of the promoter regions of p16 genes determined by methylation-specific PCR. In contrast, unmethylated p16 genes were increased with the treatment of T-47D cells with $20{\mu}M$ of caffeic acid or chlorogenic acid for 6 days. Treatment of T-47D cells with 5, 20 or $50{\mu}M$ of EGCG increased the unmethylation status of p16 gene up to 100%, and the methylation-specific bands of this gene were decreased up to 50% in a concentration-dependent manner. The finding of present study demonstrated that coffee polyphenols and EGCG have strong inhibitory effects of the cellular DNA methylation process through increased formation of S-adenosyl-homocysteine(SAH) during the catechol-O-methyltransferase (COMT)- mediated O-methylation of these dietary chemicals or an direct inhibition of the DNA methyltransferases. In conclusion, various dietary polyphenols could reverse the methylation status of p16 gene in human breast T-47D cells.
Purpose : Various proteins encoded in the early region 3 (E3) of adenoviruses protect cells from being killed by cytotoxic T cells and death-inducing cytokines. We sought to find out whether the genetic heterogeneity of the E3 gene might contribute to the molecular diversity of adenoviruses. Methods : Sequences in the E3 region were analyzed for 14 adenovirus type 3 (Ad3) strains that were isolated from children with lower respiratory tract infections in the Seoul National University Children's Hospital during the period 1991-2000. Full-length adenoviral DNA was purified from the infected A549 cell lysates using a modified Hirt procedure. Results : There was 98% homology between 14 Korean Ad3 strains with a reference strain (M15952). Homology within the Korean Ad3 strains was 98.7%. Variation was found in the region of transcripts 20.1 kDa, 20.6 kDa, truncated 7.7 kDa, 10.3 kDa, 14.9 kDa, and 15.3 kDa. In particular, all 14 Korean strains showed a missense single point mutation at the start codon of the truncated 7.7 kDa. In addition, a deletion was found in the truncated 7.7 kDa region by 58 base pairs in 10 strains and 94 base pairs in 4 strains. Variations in amino acids were observed in the receptor internalization and degradation complex (10.3 kDa/14.9 kDa) which stimulates the clearance from the cell surface and subsequent degradation of the receptors for the Fas ligand and TRAIL, while no variations were observed in another immunoregulatory transcript, 19 kDa. Conclusion : Sequence analysis of the immunoregulatory region of adenovirus E3 shows that genetic heterogeneities are related to genome type patterns.
The objective of this study was to evaluate the anti-inflammation effect of extract of Carthamus tinctorious seed, on skin obtained from Gyeong buk, Korea. Regulatory mechanisms of cytokines and nitric oxide (NO) involved in immunological activity of Raw 264.7 cells. Tested cells were pretreated with 70% ethanol extracted of Carthamus tinctorious seed and further cultured for an appropriated time after the addition of lipopolyssacharide (LPS). During the entire experimental period, 5, 10, 25 and 50 ${\mu}g/ml$ of Carthamus tinctorious seed showed no cytotoxicity. In these concentrations, ethyl acetate layer of ethanol extracted Carthamus tinctorius seed (CT-E/E) inhibited the production of NO and prostaglandin $E_2$ ($PGE_2$), tumor necorsis factor-a (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$), interleukin-6 (IL-6) expression of inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2). At a 50 ${\mu}g/ml$ level of CT-E/E, $PGE_2$, iNOS and COX-2 inhibition activity were shown 60%, 38%, and 42%, respectively. In addition, CT-E/E reduced the release of inflammatory cytokines including TNF-${\alpha}$, IL-$1{\beta}$ and IL-6. These results suggest that Carthamus tinctorious seed extracts may be a potential anti-inflammatory therapeutic agent due to the significant effects on inflammatory factors.
Seunghyun Ahn;Jung Yeon Lee;Eunbi Hong;Jiyun Kim;Won Seok Jeong;Kown Ki Moon;CheongTaek Kim;Jiha Sung;Seyeon Park
Journal of Applied Biological Chemistry
/
v.65
no.4
/
pp.269-275
/
2022
Betula platyphylla extract includes various materials which showed biological activity such as terpenoids. For this reason, Betula platyphylla extract has been used to alleviate inflammation. In this study, extract of Betula platyphylla was obtained and purified using several solvents and evaluated whether they showed effect on prevention of hair loss. Cell cytotoxicity assay was performed to investigate the effect of extracts on cell proliferation. Western blotting was performed to observe the changes in expression of several related growth factors such as β-catenin, VEGF, IGF1, and cyclin D. Also, 5-α-reductase activity was measured. The ethyl acetate extract was divided into four partial extracts and named as H3-1, H3-2, H3-3, and H3-4. The H3-2 extract showed proliferation activity of human derma papilla cell and increased the protein expression of several related growth factors such as β-catenin, VEGF, IGF1, and cyclin D, comparable to the effect of Ethyl 3,4,5-Trimethoxy Benzoate (ETB)and Lupeol (LPO). Moreover, we found that the fraction H3 was shown to decrease 5-α-reductase activity while ETB and LPO had no significant effect on 5-α-reductase activity.
Inflammation by the innate immune system is a protective mechanism of the organism against infection-mediated environmental factors. It is also responsible for the pathogenesis of various human diseases, including the progression of cancer. Lichens are receiving increasing attention as a source of bioactive molecules with therapeutic potential for a variety of diseases. Additionally, the antioxidant, anti-inflammatory, and anticancer potential of lichen and its secondary metabolites have been widely reported. However, the underlying mechanism is still unknown. In the present study, to investigate molecular mechanisms of anti-inflammation and anti-cancer activity in the Antarctic lichen, Usnea aurantiaco-atra, methanol extract of Usnea aurantiaco-atra (MEUS) was used in vitro assays in RAW 264.7 macrophages cell and HCT116 colon cancer cells. Based on our data, MEUS had the anti-inflammatory activity through the modulation of main inflammatory indicators such as COX-2, IL-6, iNOS, TNF-α and NO production in a concentration-dependent manner. In addition, we observed that MEUS had cytotoxic activity against HCT116 colon cancer cells in a concentration-dependent manner, leading to a significantly reduced proliferation of the cancer cells through apoptotic induction by activating caspase-3. Taken together, this work firstly reported the anti-inflammatory and anti-cancer activities of an Antarctic lichen, Usnea aurantiaco-atra, and MEUS may provide a new insight into the molecular mechanisms underlying a link between inflammation and cancer.
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