• Title/Summary/Keyword: inhibitor p21

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Characterization of Polyphenoloxidase extracted from Solanum tuberosum Jasim

  • Jang, Jae-Won;Ma, Yu-Hyun;Shin, Ju-Mi;Song, Kyung-Bin
    • Food Science and Biotechnology
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    • v.14 no.1
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    • pp.117-122
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    • 2005
  • Polyphenoloxidase (PPO) was extracted from Solanum tuberosum Jasim by various chromatographic methods and was subsequently purified and characterized. PPO was purified upto 78-fold from the crude extract. SDS-PAGE profile of the enzyme showed a major subunit of PPO with molecular weight of 40 kDa. The optimum pH and temperature for the maximum activity of PPO was 6.5 and $25^{\circ}C$, respectively. The enzyme was found to be quite stable between 10 and $40^{\circ}C$, whereas it was almost inactivated at $70^{\circ}C$ when incubated for 30 min. Substrate specificity study indicated that catechol was the most suitable substrate for PPO isolated from purple-fleshed potato with a $K_m$ value of 21.1 mM. The most effective inhibitor was ascorbic acid, followed by L-cysteine, citric acid, EDTA, and boric acid. Studies on the effect of metal ion on PPO activity showed that magnesium and copper were inhibitory, while iron and zinc ions increased the activity of PPO.

Anticancer and Antimicrobial Activities of 13(E)-labd-13-ene-8α,15-diol from Brachyglottis monroi

  • Kim, Jong-Im;Choi, Hwa-Jung;Lee, Jae-Sook
    • Journal of Applied Biological Chemistry
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    • v.56 no.1
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    • pp.49-51
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    • 2013
  • In a previous study, we reported that 13(E)-labd-13-ene-$8{\alpha}$,15-diol (13E) possesses antiviral and anticancer activities. In this study, the anticancer and antimicrobial activities of 13(E) were evaluated against 4 cancer cell lines and 6 bacteria. 13(E) showed inhibitory effect on a variety of cancer cell lines. The $IC_{50}$ values was 8.3-21.3 ${\mu}g/mL$. 13(E) was the most effective growth inhibitor of murine leukaemia cell lines P388, producing approximately 8.3 ${\mu}g/mL$ of $IC_{50}$ in the cytopathic effect (CPE) method. 13(E) also inhibited the growth of the gram-positive bacteria (Staphylococcus aureus, Bacillus cereus and Listeria monocytogenes) and gram-negative bacteria (Vibrio parahaemolyticus, Escherichia coli and Salmonella enteritidis) with a range of minimum inhibitory concentration (MIC) values from 0.092 to 0.598 mg/mL and gram-negative bacteria were more sensitive to the compound (MIC, 0.092 mg/mL).

Anticancer effects of D-pinitol in human oral squamous carcinoma cells

  • Shin, Hyun-Chul;Bang, Tea-Hyun;Kang, Hae-Mi;Park, Bong-Soo;Kim, In-Ryoung
    • International Journal of Oral Biology
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    • v.45 no.4
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    • pp.152-161
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    • 2020
  • D-pinitol is an analog of 3-methoxy-D-chiro-inositol found in beans and plants. D-pinitol has anti-inflammatory, antidiabetic, and anticancer effects. Additionally, D-pinitol induces apoptosis and inhibits metastasis in breast and prostate cancers. However, to date, no study has investigated the anticancer effects of D-pinitol in oral cancer. Therefore, in this study, whether the anticancer effects of D-pinitol induce apoptosis, inhibit the epithelial-to-mesenchymal transition (EMT), and arrest cell cycle was investigated in squamous epithelial cells. D-pinitol decreased the survival and cell proliferation rates of CAL-27 and Ca9-22 oral squamous carcinoma cells in a concentration- and time-dependent manner. Evidence of apoptosis, including nuclear condensation, poly (ADP-ribose) polymerase, and caspase-3 fragmentation, was also observed. D-pinitol inhibited the migration and invasion of both cell lines. In terms of EMT-related proteins, E-cadherin was increased, whereas N-cadherin, Snail, and Slug were decreased. D-pinitol also decreased the expression of cyclin D1, a protein involved in the cell cycle, but increased the expression of p21, a cyclin-dependent kinase inhibitor. Hence, D-pinitol induces apoptosis and cell cycle arrest in CAL-27 and Ca9-22 cells, demonstrating an anticancer effect by decreasing the EMT.

Effect of different concentrations of hypotaurine on melanosis and quality of Pacific white shrimp (Penaeus vannamei) during refrigeration

  • Zhou, Jiaying;Ying, Yubin;Zhou, Yaqi;Li, Gaoshang;Hu, Yaqin
    • Fisheries and Aquatic Sciences
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    • v.25 no.4
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    • pp.231-242
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    • 2022
  • Effect of different concentrations of hypotaurine (HTU) on melanosis and quality of shrimps during 10 d storage in ice were studied. During refrigeration, the total plate count and total volatile basic nitrogen of shrimps treated with 20 g/L HTU were the lowest, and the hardness and microstructure were the best. Moreover, the score of melanoses, pH and total bile acid of shrimps treated with 20 g/L HTU were also low. Sensory evaluation showed that HTU treatment could make the shelf life of shrimps 3-4 days longer compared with the control. Based on the above physical and chemical indexes, 20 g/L HTU showed great potential as a safe inhibitor in the treatment of shrimps' melanosis.

Cornuside inhibits glucose-induced proliferation and inflammatory response of mesangial cells

  • Xiaoxin Li;Lizhong Guo;Fei Huang;Wei Xu;Guiqing Peng
    • The Korean Journal of Physiology and Pharmacology
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    • v.27 no.6
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    • pp.513-520
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    • 2023
  • Cornuside is a secoiridoid glucoside compound extracted from the fruits of Cornus officinalis. Cornuside has immunomodulatory and anti-inflammatory properties; however, its potential therapeutic effects on diabetic nephropathy (DN) have not been completely explored. In this study, we established an in vitro model of DN through treating mesangial cells (MMCs) with glucose. MMCs were then treated with different concentrations of cornuside (0, 5, 10, and 30 μM). Cell viability was determined using cell counting kit-8 and 5-ethynyl-2'-deoxyuridine assays. Levels of proinflammatory cytokines, including interleukin (IL)-6, tumor necrosis factor-α, and IL-1β were examined using enzyme-linked immunosorbent assay. Reverse transcription quantitative real-time polymerase chain reaction and Western blotting were performed to detect the expression of AKT and nuclear factor-kappa B (NF-κB)-associated genes. We found that cornuside treatment significantly reduced glucose-induced increase in MMC viability and expression of pro-inflammatory cytokines. Moreover, cornuside inhibited glucose-induced phosphorylation of AKT and NF-κB inhibitor alpha, decreased the expression of proliferating cell nuclear antigen and cyclin D1, and increased the expression of p21. Our study indicates that the anti-inflammatory properties of cornuside in DN are due to AKT and NF-κB inactivation in MMCs.

Sphingosine-1-Phosphate-Induced Migration and Differentiation of Human Mesenchymal Stem Cells to Smooth Muscle Cells (Sphingosine-1-phosphate에 의한 중간엽 줄기세포의 이동과 평활근세포로의 분화)

  • Song, Hae-Young;Shin, Sang-Hun;Kim, Min-Young;Kim, Jae-Ho
    • Journal of Life Science
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    • v.21 no.2
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    • pp.183-193
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    • 2011
  • Migration and differentiation of mesenchymal stem cells are crucial for tissue regeneration in response to injury. Sphingosine-1-phosphate (S1P) is a bioactive lipid that regulates a variety of biological processes, including proliferation, survival, differentiation and motility. In the present study, we determined the role of S1P in migration and differentiation of human bone marrow-derived mesenchymal stem cells (BMSCs). S1P stimulated migration of BMSCs in a dose- and time-dependent manner, and pre-incubation of the cells with pertussis toxin completely abrogated S1P-induced migration, suggesting involvement of Gi-coupled receptors in S1P-induced cell migration. S1P elicited elevation of intracellular concentration of $Ca^{2+}$ ($[Ca^{2+}]_i$) and pretreatment with VPC23019, an antagonist of $S1P_1/S1P_3$, blocked S1P-induced migration and increase of $[Ca^{2+}]_i$. Small interfering RNA-mediated knockdown of endogenous $S1P_1$ attenuated S1P-induced migration of BMSCs. Furthermore, S1P treatment induced expression of $\alpha$-smooth muscle actin ($\alpha$-SMA), a smooth muscle marker, and pretreatment with VPC23019 abrogated S1P-induced $\alpha$-SMA expression. S1P induced phosphorylation of p38 mitogen-activated protein kinase (MAPK), and pretreatment of cells with SB202190, an inhibitor of p38 MAPK, or adenoviral overexpression of a dominant-negative mutant of the p38 MAPK blocked S1P-induced cell migration and $\alpha$-SMA expression. Taken together, these results suggest that S1P stimulates migration and smooth muscle differentiation of BMSCs through an $S1P_1$-p38 MAPK-dependent mechanism.

A Novel Suberoylanilide Hydroxamic Acid Histone Deacetylase Inhibitor Derivative, N25, Exhibiting Improved Antitumor Activity in both Human U251 and H460 Cells

  • Zhang, Song;Huang, Wei-Bin;Wu, Li;Wang, Lai-You;Ye, Lian-Bao;Feng, Bing-Hong
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.10
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    • pp.4331-4338
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    • 2014
  • $N^1$-(2, 5-dimethoxyphenyl)-$N^8$-hydroxyoctanediamide (N25) is a novel SAHA cap derivative of HDACi, with a patent (No. CN 103159646). This invention is a hydroxamic acid compound with a structural formula of $RNHCO(CH_2)6CONHOH$ (wherein R=2, 5dimethoxyaniline), a pharmaceutically acceptable salt which is soluble. In the present study, we investigated the effects of N25 with regard to drug distribution and molecular docking, and anti-proliferation, apoptosis, cell cycling, and $LD_{50}$. First, we designed a molecular approach for modeling selected SAHA derivatives based on available structural information regarding human HDAC8 in complex with SAHA (PDB code 1T69). N25 was found to be stabilized by direct interaction with the HDAC8. Anti-proliferative activity was observed in human glioma U251, U87, T98G cells and human lung cancer H460, A549, H1299 cells at moderate concentrations ($0.5-30{\mu}M$). Compared with SAHA, N25 displayed an increased antitumor activity in U251 and H460 cells. We further analyzed cell death mechanisms activated by N25 in U251 and H460 cells. N25 significantly increased acetylation of Histone 3 and inhibited HDAC4. On RT-PCR analysis, N25 increased the mRNA levels of p21, however, decreased the levels of p53. These resulted in promotion of apoptosis, inducing G0/G1 arrest in U251 cells and G2/M arrest in H460 cells in a time-dependent and dose-dependent manner. In addition, N25 was able to distribute to brain tissue through the blood-brain barrier of mice ($LD_{50}$: 240.840mg/kg). In conclusion, our findings demonstrate that N25 will provide an invaluable tool to investigate the molecular mechanism with potential chemotherapeutic value in several malignancies, especially human glioma.

Pasteurization Efficiency and Physico-chemical Changes of Soymilk HTST Pasteurized Using Microwaves (두유의 마이크로파 고온단시간 살균시 살균효과 및 이화학적 성분 변화)

  • Kim, Suk-Shin;Lee, Joo-Hee
    • Korean Journal of Food Science and Technology
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    • v.31 no.5
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    • pp.1196-1202
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    • 1999
  • This work was to determine the microbial and physico-chemical changes of HTST-pasteurized soymilk using microwave energy. Soymilk was HTST pasteurized$(at\;90^{\circ}C\;for\;20\;sec)$ by three methods: by heating in a stainless steel tube immersed in a hot water bath(MP0), by heating in a microwave cavity to a defiled temperature and then holding in a hot water bath(MP1), and by both heating and holding in a microwave cavity(MP2). The microbial quality based on the total plate count was in the order of MP0, MP2 and MP1. The three samples pasteurized by different methods showed the similar microbial quality with respect to the coliform count, psychrotrophic bacterial count and phosphatase activity. The destruction of trypsin inhibitor was in the order of MP0, MP1 and MP2. There were no significant differences in pH, titratable acidity, viscosity and vitamin $B_2$ content before and after pasteurization and among the different pasteurization methods. The similar or higher quality retention of the MP1 or MP2 supports the possibility of using microwave energy for the HTST pasteurization of soymilk and other fluid food products.

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Protein kinase C beta II upregulates intercellular adhesion molecule-1 via mitochondrial activation in cultured endothelial cells

  • Joo, Hee Kyoung;Lee, Yu Ran;Choi, Sunga;Park, Myoung Soo;Kang, Gun;Kim, Cuk-Seong;Jeon, Byeong Hwa
    • The Korean Journal of Physiology and Pharmacology
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    • v.21 no.4
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    • pp.377-384
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    • 2017
  • Activation of protein kinase C (PKC) is closely linked with endothelial dysfunction. However, the effect of $PKC{\beta}II$ on endothelial dysfunction has not been characterized in cultured endothelial cells. Here, using adenoviral $PKC{\beta}II$ gene transfer and pharmacological inhibitors, the role of $PKC{\beta}II$ on endothelial dysfucntion was investigated in cultured endothelial cells. Phorbol 12-myristate 13-acetate (PMA) increased reactive oxygen species (ROS), p66shc phosphorylation, intracellular adhesion molecule-1, and monocyte adhesion, which were inhibited by $PKC{\beta}i$ (10 nM), a selective inhibitor of $PKC{\beta}II$. PMA increased the phosphorylation of CREB and manganese superoxide dismutase (MnSOD), which were also inhibited by $PKC{\beta}i$. Gene silencing of CREB inhibited PMA-induced MnSOD expression, suggesting that CREB plays a key role in MnSOD expression. Gene silencing of $PKC{\beta}II$ inhibited PMA-induced mitochondrial ROS, MnSOD, and ICAM-1 expression. In contrast, overexpression of $PKC{\beta}II$ using adenoviral $PKC{\beta}II$ increased mitochondrial ROS, MnSOD, ICAM-1, and p66shc phosphorylation in cultured endothelial cells. Finally, $PKC{\beta}II$-induced ICAM-1 expression was inhibited by Mito-TEMPO, a mitochondrial ROS scavenger, suggesting the involvement of mitochondrial ROS in PKC-induced vascular inflammation. Taken together, the results suggest that $PKC{\beta}II$ plays an important role in PMA-induced endothelial dysfunction, and that the inhibition of $PKC{\beta}II$-dependent p66shc signaling acts as a therapeutic target for vascular inflammatory diseases.

Dichloroacetate Inhibits the Proliferation of a Human Anaplastic Thyroid Cancer Cell Line via a p53-independent Pathway (Dichloroacetate의 p53 비의존적 경로를 통한 인간 역분화 갑상선 암세포주의 성장억제 효과)

  • KC, Yam Bahadur;Poudel, Sunil;Jeon, Eon Ju;Shon, Ho Sang;Byun, Sung June;Jeoung, Nam Ho
    • Journal of Life Science
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    • v.28 no.12
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    • pp.1469-1476
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    • 2018
  • Occurrence of the Warburg effect in solid tumors causes resistance to cancer chemotherapy, and targeting energy metabolisms such as aerobic glycolysis is a potential strategy for alternative treatment. Dichloroacetate (DCA), an inhibitor of pyruvate dehydrogenase kinase (PDK), shifts glucose metabolism from aerobic glycolysis to oxidative phosphorylation (OxPhos) in many cancers. In this study, we investigated the anticancer effect of DCA on a human anaplastic thyroid cancer (ATC) cell line, 8505C. We found that DCA selectively inhibits cell proliferation of the 8505C line but not of a normal thyroid line. In 8505C, the cell cycle was arrested at the G1/S phase with DCA treatment as a result of decreased antiapoptotic proteins such as $HIF1{\alpha}$, PDK1, and Bcl-2 and increased proapoptotic proteins such as Bax and p21. DCA treatment enhanced the production of reactive oxygen species which consequently induced cell cycle arrest and apoptosis. Interestingly, DCA treatment not only reduced lactate production but also increased the expression of sodium-iodine symporter, indicating that it restores the OxPhos of glucose metabolism and the iodine metabolism of the ATC. Taken together, our findings suggest that PDK inhibitors such as DCA could be useful anticancer drugs for the treatment of ATC and may also be helpful in combination with chemotherapy and radiotherapy.