• Title/Summary/Keyword: Bcl I

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Growth Inhibition and Induction of Apoptosis in Human Bladder Cancer Cells Induced by Fermented Citrus Kombucha (감귤 콤부차 발효액의 인체 방광암세포에 대한 성장억제와 Apoptosis에 미치는 영향)

  • Kim, Chung-I;Shin, Seung-Shick;Park, Sung-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.10
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    • pp.1422-1429
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    • 2016
  • Kombucha is a slightly sour beverage fermented by symbiotic micro-organisms, including bacteria and yeasts. In this study, we examined the biological activities of citrus Kombucha (CK) produced by addition of citrus extract to original Kombucha (K). After fermentation for 10 days, radical scavenging activity examined by ABTS and DPPH assays increased by approximately 20% compared to that of K. Moreover, content of total phenolic compounds significantly increased by 60% compared to that of K. Cell proliferation assays utilizing MTT showed that CK treatment significantly inhibited growth of bladder cancer cells, T-24 and 5637, in a dose-dependent manner with $IC_{50}$ values of 4 and 7 mg/mL, respectively. Annexin V staining showed that CK treatment led to apoptosis of cells in a dose-dependent manner. T-24 cells were more sensitive to CK treatment than 5637 cells, as 8 mg/mL of CK resulted in 97% apoptosis of T-24 cells. Western blotting showed that CK treatment led to up-regulation of apoptotic proteins, including caspases-3, -8, -9, and PARP, in bladder cells not in K-treated cells. Taken together, these results demonstrate that CK may be developed as a functional beverage.

Anti-inflammatory and Anti-allergic Effect of Extractsfrom Organic Soybean (유기농 콩 추출물의 항염증 및 항알레르기 효과)

  • Chung, Eun-Kyung;Seo, Eun-Hye;Park, Jun-Ho;Shim, Hye-Rim;Kim, Kyung-Hee;Lee, Byung-Ryong
    • Korean Journal of Organic Agriculture
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    • v.19 no.2
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    • pp.245-253
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    • 2011
  • This study was carried out to investigate the biological effects of organic soybean extracts. Cellular and molecular analysis was performed to determine anti-oxidative, anti-inflammatory, anti-apoptotic, and anti-inflammatory effects of soybean extracts. First, we obtained various solvent extracts of soybean such as water, ethanol, and methanol. Molecular and cellular analysis were performed with 0.1 mg/ml concentration of each solvent extracts. The results of anti-oxidative, antiinflammatory and anti-apoptotic effects of organic soybean extracts were prominent. However, organic soybean extracts were not observed in anti-allergic effects determined by releasing histamine from rat mast cell line, RBL-2H3. Conclusively, organic soybean suppress inflammatory responses. In addition, organic soybean could be applied as a functional food ingredient for treatment of chronic inflammation, asthma, and atopic dermatitis with enhanced anti-inflammatory activities.

Semaphoring mAb: a New Guide in RIT in Inhibiting the Proliferation of Human Skin Carcinoma

  • Liu, Yuan;Ma, Jing-Yue;Luo, Su-Ju;Sun, Chen-Wei;Shao, Li-Li;Liu, Quan-Zhong
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.3
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    • pp.941-945
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    • 2015
  • Semaphoring is a transmembrane receptor which participates in many cytokine-mediated signal pathways that are closely related to the angiogenesis, occurrence and development of carcinoma. The present study was designed to access the effect of mono-antibody (mAb) guided radioimmunotherapy (RIT) on skin carcinoma and investigate the potential mechanisms. Semaphoring mAb was acquired from mice (Balb/c), purified with rProtein A column; purity, concentration and activity were tested with SDS-PAGE and indirect ELISA; specificity and expression on the cutanuem carcinoma line and tissue were tested by Western blotting; morphology change was assessed by microscopy. MTT assay and colony inhibition tests were carried out to test the influence on the proliferation of tumor cells; Western blotting was also carried out for expression of apoptosis-associated (caspase-3, Bax, Bcl-2) and proliferation-related (PI3K, p-Akt, Akt, p-ERK1/2, ERK1/2) proteins and analyse the change in signal pathways (PI3K/Akt and MEK/ERK). The purity of purified semaphorin mAb was 96.5% and the titer is about $1{\times}10^6$. Western blotting showed semaphoring mAb to have specifically binding stripes with semaphoring b1b2 protein, B16F10, and A431 cells at 39KDa, 100KDa and 130KDa, respectively. Positive expression was detected both in cutanuem carcinoma line and tissue and it mostly located in cell membranes. MMT assay revealed dose-relate and time-relate inhibitory effect of semaphorin mAb on A431 and B16F10. Colony inhibition tests also showed dose-relate inhibitory effects. Western blotting demonstrated the expression of apoptosis and proliferation-related protein and changes in signal pathway. In conclusion, we demonstrated that semaphorin is highly expressed on the tumor cell-surfaces and RIT with semaphorin mAb has effect in i nhibiting proliferation and accelerating apoptosis of tumor cells.

Improvement Effect of Sprout of Coix lacryma-jobi var. mayuen Stapf Water Extract on DSS-Induced Ulcerative Colitis in Mice. (DSS로 궤양성 대장염 유발된 동물모델에서 의이아(薏苡芽) 열수 추출물의 개선 효과)

  • Kim, Min Ju;Shin, Mi-Rae;Lee, Jin A;Park, Soon-Ae;Park, Hae-Jin;Lee, Jeong Hoon;Roh, Seong-Soo
    • The Korea Journal of Herbology
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    • v.35 no.6
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    • pp.21-28
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    • 2020
  • Objectives : The objective of this study was to investigate the improvement effect of Sprout of Coix lacryma-jobi var. mayuen Stapf water extract (SC) on the dextran sulfate sodium (DSS)-induced ulcerative colitis mice. Methods : The antioxidant activity of SC was measured through total polyphenol and total flavonoid content in vitro. The experiment was conducted with seven-week-old male Balb/c mice. After 1 week adaptation, acute colitis was induced by oral administration of 5% DSS dissolved in drinking water, for 7 days. And normal mice received drinking water without DSS throughout the entire experimental period. For each experiment, the mice were divided into 4 groups and 24 colitis mice were arbitrarily allocated into 3 groups (n = 8/group); Normal group, Control group, SC 100 mg/kg treated group (SCL), SC 200 mg/kg treated group (SCH). Serum and colon tissues were collected after one weeks of drug administration. Results : ROS levels, ONOO- levels, AST, and ALT in serum were decreased in SC treated groups compared to the control group. Western blotting measurements of Nrf2, HO-1, SOD, catalase, GPx-1/2, IL-4, IL-10, and Bcl2 showed that the SC treated groups was increased compared to the Control group. Also, western blot measurements of NF-κBp65, p-IκBα, COX-2, iNOS, TNF-α, IL-1β, Bax, and Caspase-3 showed that the SC treated groups was reduced compared to the Control group. Conclusion : Taken together, these results suggest that SC treatment can attenuate the DSS-induced colitis though inhibiting NF-κB pathway and enhancing Nrf2 pathway. Therefore, SC was the potential to be used as a natural therapeutic drug.

Anti-inflammatory and anti-allergic effect of soybean extracts produced by organic cultivation (유기농 대두 추출물의 항염증 및 항알레르기 효과)

  • Chung, Eun-Kyung;Seo, Eun-Hye;Park, Jun-Ho;Kim, Young-Nam;Kim, Kyung-Hee;Lee, Byung-Ryong
    • Proceedings of the Korean Society of Organic Agriculture Conference
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    • 2009.12a
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    • pp.103-113
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    • 2009
  • This present study was carried out to investigate the biological effects of soybean extracts comparing organic and conventional cultivation. Cellular and molecular analysis was performed to determine anti-oxidative, anti-inflammatory, anti-apoptotic, and anti-inflammatory effects of both soybean extracts. First, we obtained various solvent extracts of soybeans such as water, ethanol, and methanol. Molecular and cellular analysis were performed with 0.1 mg/ml concentration of each solvent extracts. The results of anti-oxidative, anti-inflammatory and anti-apoptotic effects of organic cultivated soybean extracts were prominent than conventional cultivated soybean extracts. However, discrepancy between organic and conventional cultivated soybean extracts was not observed in anti-allergic effects determined by releasing histamine from rat mast cell line, RBL-2H3. Conclusively, organic cultivated soybeans have stronger effects than conventional cultivated soybeans in suppression of inflammation. In addition, organic soybeans could be applied as a functional food ingredient for treatment of chronic inflammation, asthma, and atopic dermatitis with enhanced anti-inflammatory activities.

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The Antitumor Effects of Selenium Compound $Na_5SeV_5O_{18}{\cdot}3H_2O$ in K562 Cell

  • Yang, Jun-Ying;Wang, Zi-Ren
    • Archives of Pharmacal Research
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    • v.29 no.10
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    • pp.859-865
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    • 2006
  • With an approach to study the anti-tumor effects and mechanism of selenium compound, we investigated the anti-tumor activity and mechanism of $Na_5SeV_5O_{18}{\cdot}3H_2O$ (NaSeVO) in K562 cells. The results showed that $0.625{\sim}20\;mg/L$ NaSeVO could significantly inhibit the proliferation of K562 cells in vitro in a time- and concentration-dependent manner as determined by microculture tetrazolium (MTT) assay, the IC50 values were 14.41 (4.45-46.60) and 3.45 (2.29-5.22) mg/L after 48 hand 72 h treatment with NaSeVO respectively. In vivo experiments demonstrated that i.p. administration of 5, 10 mg/kg NaSeVO exhibited an significant inhibitory effect on the growth of transplantation tumor sarcoma 180 (S180) and hepatoma 22 (H22) in mice, with inhibition rate 26.8% and 58.4% on S180 and 31.3% and 47.4% on H22, respectively. Cell cycle studies indicated that the proportion of G0/G1 phase was increased at 2.5 mg/L while decreased at 10 mg/L after treatment for 24, 48 h. Whereas S phase was decreased at 2.5-5 mg/L and markedly increased at 10 mg/L after treatment for 48 h. After treatment for 24 h, 10 mg/L NaSeVO also markedly increased S and G2/M phases. Take together, the result clearly showed that NaSeVO markedly increased S and G2/M phases at 10 mg/L. The study of immunocytochemistry showed that the expression bcl-2 is significantly inhibited by 10 mg/L NaSeVO, and bax increased. Morphology observation also revealed typical apoptotic features. NaSeVO also significantly caused the accumulation of $Ca^{2+}$ and $Mg^{2+}$, reactive oxygen species (ROS) and the reduction of pH value and mitochondrial membrane potential in K562 cells as compared with control by confocal laser scanning microscope. These results suggest that NaSeVO has anti-tumor effects and its mechanism is attributed partially to apoptosis induced by the elevation of intracellular $Ca^{2+}$, $Mg^{2+}$ and ROS concentration, and a reduction of pH value and mitochondria membrane potential (MMP).

Induction of Forkhead Class box O3a and apoptosis by a standardized ginsenoside formulation, KG-135, is potentiated by autophagy blockade in A549 human lung cancer cells

  • Yao, Chih-Jung;Chow, Jyh-Ming;Chuang, Shuang-En;Chang, Chia-Lun;Yan, Ming-De;Lee, Hsin-Lun;Lai, I-Chun;Lin, Pei-Chun;Lai, Gi-Ming
    • Journal of Ginseng Research
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    • v.41 no.3
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    • pp.247-256
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    • 2017
  • Background: KG-135, a standardized formulation enriched with Rk1, Rg3, and Rg5 ginsenosides, has been shown to inhibit various types of cancer cells; however, the underlying mechanisms are not fully understood. In this study, we explored its effects in A549 human lung cancer cells to investigate the induction of Forkhead Class box O3a (FOXO3a) and autophagy. Methods: Cell viability was determined by sulforhodamine B staining. Apoptosis and cell cycle distribution were analyzed using flow cytometry. The changes of protein levels were determined using Western blot analysis. Autophagy induction was monitored by the formation of acidic vesicular organelles stained with acridine orange. Results: KG-135 effectively arrested the cells in G1 phase with limited apoptosis. Accordingly, a decrease of cyclin-dependent kinase-4, cyclin-dependent kinase-6, cyclin D1, and phospho-retinoblastoma protein, and an increase of p27 and p18 proteins were observed. Intriguingly, KG-135 increased the tumor suppressor FOXO3a and induced the accumulation of autophagy hallmark LC3-II and acidic vesicular organelles without an increase of the upstream marker Beclin-1. Unconventionally, the autophagy adaptor protein p62 (sequestosome 1) was increased rather than decreased. Blockade of autophagy by hydroxychloroquine dramatically potentiated KG-135-induced FOXO3a and its downstream (FasL) ligand accompanied by the cleavage of caspase-8. Meanwhile, the decrease of Bcl-2 and survivin, as well as the cleavage of caspase-9, were also drastically enhanced, resulting in massive apoptosis. Conclusion: Besides arresting the cells in G1 phase, KG-135 increased FOXO3a and induced an unconventional autophagy in A549 cells. Both the KG-135-activated extrinsic FOXO3a/FasL/caspase-8 and intrinsic caspase-9 apoptotic pathways were potentiated by blockade of autophagy. Combination of KG-135 and autophagy inhibitor may be a novel strategy as an integrative treatment for cancers.

Baicalein induces cell death in Human Lung Carcinoma A549 Cells: Role of Apoptosis and Autophagy pathway (인체폐암 A549 세포에서 Baicalein에 의한 세포사멸 유도: Apoptosis와 Autophagy 경로의 역할)

  • Kim, Chul Hwan;Hwang, Buyng Su;Jeong, Yong Tae;Kim, Min-Jin;Shin, Su Young;Oh, Young Taek;Eom, Jung Hye;Lee, Seung Young;Choi, Kyung Min;Cho, Pyo Yun;Jeong, Jin-Woo
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.112-112
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    • 2019
  • Baicalein is one of the main flavonoids derived from roots of Scutellaria baicalensis Georgi, a traditional Oriental medicine. Although baicalein has high antitumor effect on several human carcinomas, the mechanism responsible for this property is not unclear. In this study, the data revealed that baicale-ininduced growth inhibition was associated with the induction of apoptosis connecting with cytochrome c release, down-regulation of anti-apoptotic Bcl-xl and increased the percentage of cells with a loss of mitochondria membrane permeabilization. Baicalein also induced the proteolytic activation of caspases and cleavage of PARP; however, blockage of caspases activation by z-VAD-fmk inhibited baicalein-induced apoptosis. In addition, baicalein enhanced the formation of autophagosomes and up-regulated LC3-II/LC3-I ratio. Interestingly, the pretreatment of bafilomycin A1 recovered baicalein-induced cell death suggesting that autophagy by baicalein roles as protective autophagy. Taken together, our results indicated that this flavonoid induces apoptosis and cell protective autophagy. These data means combination treatment with baicalein and autophagy inhibitor might be a promising anticancer drug.

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Allomyrina dichotoma larva extract attenuates free fatty acid-induced lipotoxicity in pancreatic beta cells

  • Kim, Kyong;Kwak, Min-Kyu;Bae, Gong-Deuk;Park, Eun-Young;Baek, Dong-Jae;Kim, Chul-Young;Jang, Se-Eun;Jun, Hee-Sook;Oh, Yoon Sin
    • Nutrition Research and Practice
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    • v.15 no.3
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    • pp.294-308
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    • 2021
  • BACKGROUD/OBJECTIVES: Allomyrina dichotoma larva (ADL), one of the many edible insects recognized as future food resources, has a range of pharmacological activities. In a previous study, an ADL extract (ADLE) reduced the hepatic insulin resistance of high-fat diet (HFD)-induced diabetic mice. On the other hand, the associated molecular mechanisms underlying pancreatic beta-cell dysfunction remain unclear. This study examined the effects of ADLE on palmitate-induced lipotoxicity in a beta cell line of a rat origin, INS-1 cells. MATERIALS/METHODS: ADLE was administered to high-fat diet treated mice. The expression of apoptosis-related molecules was measured by Western blotting, and reactive oxidative stress generation and nitric oxide production were measured by DCH-DA fluorescence and a Griess assay, respectively. RESULTS: The administration of ADLE to HFD-induced diabetic mice reduced the hyperplasia, 4-hydroxynonenal levels, and the number of apoptotic cells while improving the insulin levels compared to the HFD group. Treatment of INS-1 cells with palmitate reduced insulin secretion, which was attenuated by the ADLE treatment. Furthermore, the ADLE treatment prevented palmitate-induced cell death in INS-1 cells and isolated islets by reducing the apoptotic signaling molecules, including cleaved caspase-3 and PARP, and the Bax/Bcl2 ratio. ADLE also reduced the levels of reactive oxygen species generation, lipid accumulation, and nitrite production in palmitate-treated INS-1 cells while increasing the ATP levels. This effect corresponded to the decreased expression of inducible nitric oxide synthase (iNOS) mRNA and protein. CONCLUSIONS: ADLE helps prevent lipotoxic beta-cell death in INS-1 cells and HFD-diabetic mice, suggesting that ADLE can be used to prevent or treat beta-cell damage in glucose intolerance during the development of diabetes.