• Title/Summary/Keyword: Cycling Time

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Antitumor Activity of Histone Deacetylase Inhibitor Trichostatin A in Osteosarcoma Cells

  • Cheng, Dong-Dong;Yang, Qing-Cheng;Zhang, Zhi-Chang;Yang, Cui-Xia;Liu, Yi-Wen
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.4
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    • pp.1395-1399
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    • 2012
  • Background: Histone deacetylase (HDAC) inhibitors have been reported to induce cell growth arrest, apoptosis and differentiation of tumor cells. The present study aimed to examine the effects of trichostatin A (TSA), one such inhibitor, on the cell cycle, apoptosis and invasiveness of osteosarcoma cells. Methods: MG-63 cells were treated with TSA at various concentrations. Then, cell growth and apoptosis were determined by 3-(4, 5-dimethyl-2-thiazolyl)-2H-tetrazolium bromide (MTT) and TUNEL assays, respectively; cell cycling was assessed by flow cytometry; invasion assays were performed with the transwell Boyden Chamber system. Results: MTT assays revealed that TSA significantly inhibited the growth of MG-63 cells in a concentration and time dependent manner. TSA treated cells demonstrated morphological changes indicative of apoptosis and TUNEL assays revealed increased apoptosis of MG-63 cells after TSA treatment. Flow cytometry showed that TSA arrested the cell cycle in G1/G2 phase and annexin V positive apoptotic cells increased markedly. In addition, the invasiveness of MG-63 cells was inhibited by TSA in a concentration dependent manner. Conclusion: Our findings demonstrate that TSA inhibits the proliferation, induces apoptosis and inhibits invasiveness of osteosarcoma cells in vitro. HDAC inhibitors may thus have promise to become new therapeutic agents against osteosarcoma.

PEDOT:PSS Enhanced Electrochemical Capacitive Performance of Graphene-Templated δ-MnO2

  • Sinan, Neriman;Unur, Ece
    • Journal of Electrochemical Science and Technology
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    • v.11 no.1
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    • pp.50-59
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    • 2020
  • Birnessite-type manganese dioxide (δ-MnO2) with hierarchical micro-/mesoporosity was synthesized via sacrificial graphene template approach under mild hydrothermal conditions for the first time. Graphene template was obtained by a surfactant (cetyltrimethylammonium bromide, CTAB) assisted liquid phase exfoliation (LPE) in water. A thin PEDOT:PSS (poly (3,4-ethylene dioxythiophene): poly (styrene sulfonate)) layer was applied to improve electrical conductivity and rate capability of MnO2. The MnO2 (535 F g-1 at 1 A g-1 and 45 F g-1 at 10 A g-1) and MnO2/PEDOT:PSS nanocomposite (550 F g-1 at 1 A g-1 and 141 F g-1 at 10 A g-1) delivered electrochemical performances superior to their previously reported counterparts. An asymmetric supercapacitor, composed of MnO2/PEDOT:PSS (positive) and Fe3O4/Carbon (negative) electrodes, provided a maximum specific energy of 18 Wh kg-1 and a maximum specific power of 4.5 kW kg-1 (ΔV= 2 V, 1M Na2SO4) with 85% capacitance retention after 1000 cycles. The graphene-templated MnO2/PEDOT:PSS nanocomposite obtained by a simple and green approach promises for future energy storage applications with its remarkable capacitance, rate performance and cycling stability

Effect of Removal of Follicles through Repeated Transvaginal Follicle Aspiration on Subsequent Follicular Populations in Murrah Buffalo (Bubalus bubalis

  • Akshey, Y.S.;Palta, P.;Manik, R.S.;Vivekananad, Vivekananad;Chauhan, M.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.5
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    • pp.632-636
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    • 2005
  • This study was conducted to investigate the effects of removal of all ovarian follicles through repeated transvaginal follicle aspiration (TVFA) on the subsequent follicular populations in buffaloes. This information is crucial for determining the optimum time interval between successive aspirations for recovering oocytes from live buffaloes through Transvaginal Oocyte Retrieval (TVOR). The oestrus of cycling buffaloes (n=5) were synchronized by a single PGF injection schedule. All the ovarian follicles were removed once every 7 days for 6 weeks through TVFA, starting from Day 7 of the oestrous cycle (Day 0 = day of oestrus). The number and size of individual ovarian follicles was recorded at Day 3 and Day 5 (Day 0 = day of TVFA) through transrectal ultrasonography. The follicles were classified on the basis of their diameter as small (3-5 mm), medium (6-9 mm) and large ($\geq$10 mm). There was no difference in the number of small and medium follicles, and the number of total follicles between Day 3 and Day 5. However, the number of large follicles was significantly higher (p<0.05) at Day 5 than that at Day 3. There was a significant (p<0.05) decrease in the proportion of small follicles and an increase (p<0.05) in the proportion of large follicles from Day 3 to Day 5, with no change in the proportion of medium follicles. The number of total follicles at Day 3 or Day 5 did not differ during the 6 TVFA sessions. It can be concluded that an interval of 3 days is more suitable than that of 5 days between successive aspirations for recovering oocytes through TVOR in a twice weekly schedule and that repeated removal of follicles through TVFA does not adversely affect the number of total follicles 3 or 5 days after TVFA.

Induction of Apoptosis and Cell Cycle Arrest by Dorema Glabrum Root Extracts in a Gastric Adenocarcinoma (AGS) Cell Line

  • Jafari, Naser;Zargar, Seyed Jalal;Yassa, Narguess;Delnavazi, Mohammad Reza
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.12
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    • pp.5189-5193
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    • 2016
  • Objective: Dorema glabrum Fisch. & C.A. Mey is a perennial plant that has several curative properties. Anti-proliferative activity of seeds of this plant has been demonstrated in a mouse fibrosarcoma cell line. The aim of the present study was to evaluate cytotoxicity of D. glabrum root extracts in a human gastric adenocarcinoma (AGS) cell line and explore mechanisms of apoptosis induction, cell cycle arrest and altered gene expression in cancer cells. Materials and Methods: The MTT assay was used to evaluate IC50 values, EB/AO staining to analyze the mode of cell death, and flow cytometry to assess the cell cycle. Quantitative real-time polymerase chain reaction (qRT-PCR) amplification was performed with apoptosis and cell cycle-related gene primers, for cyclin D1, c-myc, survivin, VEGF, Bcl-2, Bax, and caspase-3 to determine alteration of gene expression. Results: Our results showed that n-hexane and chloroform extracts had greatest toxic effects on gastric cancer cells with IC50 values of $6.4{\mu}g/ml$ and $4.6{\mu}g/ml$, respectively, after 72 h. Cell cycle analysis revealed that the population of treated cells in the G1 phase was increased in comparison to controls. Cellular morphological changes indicated induction of apoptosis. In addition, mRNA expression levels of Bax and caspase-3 were increased, and of bcl-2 survivin, VEGF, c-myc and cyclin D1 were decreased. Conclusion: Our study results suggest that D. glabrum has cytotoxic effects on AGS cells, characterized by enhanced apoptosis, reduced cell viability and arrest of cell cycling.

Effect of NUCKS-1 Overexpression on Cytokine Profiling in Obese Women with Breast Cancer

  • Soliman, Nema Ali;Zineldeen, Doaa Hussein;El-Khadrawy, Osama Helmy
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.2
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    • pp.837-845
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    • 2014
  • Background: Overweight and obesity are recognized as major drivers of cancers including breast cancer. Several cytokines, including interleukin-6 (IL-6), IL-10 and lipocalin 2 (LCN2), as well as dysregulated cell cycle proteins are implicated in breast carcinogenesis. The nuclear, casein kinase and cyclin-dependent kinase substrate-1 (NUCKS-1), is a nuclear DNA-binding protein that has been implicated in several human cancers, including breast cancer. Objectives: The present study was conducted to evaluate NUCKS-1 mRNA expression in breast tissue from obese patients with and without breast cancer and lean controls. NUCKS-1 expression was correlated to cytokine profiles as prognostic and monitoring tools for breast cancer, providing a molecular basis for a causal link between obesity and risk. Materials and Methods: This study included 39 females with breast cancer (G III) that was furtherly subdivided into two subgroups according to cancer grading (G IIIa and G IIIb) and 10 control obese females (G II) in addition to 10 age-matched healthy lean controls (G I). NUCKS-1 expression was studied in breast tissue biopsies by means of real-time PCR (RT-PCR). Serum cytokine profiles were determined by immunoassay. Lipid profiles and glycemic status as well as anthropometric measures were also recorded for all participants. Results: IL-6, IL-12 and LCN2 were significantly higher in control obese and breast cancer group than their relevant lean controls (p<0.05), while NUCKS-1 mRNA expression was significantly higher in the breast cancer group compared to the other groups (p<0.05). Significant higher levels of IL-6, IL-12, and LCN2 as well as NUCKS-1 mRNA levels were reported in G IIIb than G IIIa, and positively correlated with obesity markers in all obese patients. Conclusions: Evaluation of cytokine levels as well as related gene expression may provide a new tool for understanding interactions for three axes of carcinogenesis, innate immunity, inflammation and cell cycling, and hope for new strategies of management.

Effects of MicroRNA-106 on Proliferation of Gastric Cancer Cell through Regulating p21 and E2F5

  • Yao, Yong-Liang;Wu, Xiao-Yang;Wu, Jian-Hong;Gu, Tao;Chen, Ling;Gu, Jin-Hua;Liu, Yun;Zhang, Qing-Hui
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.5
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    • pp.2839-2843
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    • 2013
  • Objective: To investigate the effects of miR-106b on malignant characteristics of gastric cancer cells, and explore possible mechanisms. Methods: Expression of miR-106b, p21 and E2F was determined by real-time PCR. Transfection with miR-106b mimics was conducted, and gastric cancer cells with miR-106b overexpression were obtained. Cells transfected with mimic mutants and those without transfection served as negative and blank controls, respectively. Flow cytometry and transwell assays were adopted to detect the effects of miR-106b overexpression on cell cycle, migration and invasion of gastric cancer cells. Results:. The expression of miR- 106b in gastric cancer cells was significantly higher than that in normal gastric mucosa cells. Furthermore, the expression level of miR-106b rose according to the degree of malignacy among the three GC cell strains (MKN- 45 > SGC-7901 > MKN-28). Overexpression of miR-106b shortened the G0/G1 phase and accelerated cell cycle progression, while reducing p21 and E2F5, without any significant effects on the capacity for migration and invasion of gastric cancer cells. Conclusions: miR-106b may promote cell cycling of gastric cancer cells through regulation of p21 and E2F5 target gene expression.

RNAi-based Knockdown of Multidrug Resistance-associated Protein 1 is Sufficient to Reverse Multidrug Resistance of Human Lung Cells

  • Shao, Shu-Li;Cui, Ting-Ting;Zhao, Wei;Zhang, Wei-Wei;Xie, Zhen-Li;Wang, Chang-He;Jia, Hong-Shuang;Liu, Qian
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.24
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    • pp.10597-10601
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    • 2015
  • Up-regulation of multidrug resistance-associated protein 1 (MRP1) is regarded as one of the main causes for multidrug resistance (MDR) of tumor cells, leading to failure of chemotherapy-based treatment for a multitude of cancers. However, whether silencing the overexpressed MRP1 is sufficient to reverse MDR has yet to be validated. This study demonstrated that RNAi-based knockdown of MRP1 reversed the increased efflux ability and MDR efficiently. Two different short haipin RNAs (shRNAs) targeting MRP1 were designed and inserted into pSilence-2.1-neo. The shRNA recombinant plasmids were transfected into cis-dichlorodiamineplatinum-resistant A549 lung (A549/DDP) cells, and then shRNA expressing cell clones were collected and maintained. Real time PCR and immunofluorescence staining for MRP1 revealed a high silent efficiency of these two shRNAs. Functionally, shRNA-expressing cells showed increased rhodamine 123 retention in A549/DDP cells, indicating reduced efflux ability of tumor cells in the absence of MRP1. Consistently, MRP1-silent cells exhibited decreased resistance to 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) and DDP, suggesting reversal of MDR in these tumor cells. Specifically, MRP1 knockdown increased the DDP-induced apoptosis of A549/DDP cells by increased trapping of their cell cycling in the G2 stage. Taken together, this study demonstrated that RNAi-based silencing of MRP1 is sufficient to reverse MDR in tumor cells, shedding light on possible novel clinical treatment of cancers.

Down-regulation of FRα Inhibits Proliferation and Promotes Apoptosis of Cervical Cancer Cells in Vitro

  • Bai, Li-Xia;Ding, Ling;Jiang, Shi-Wen;Kang, Hui-Jie;Gao, Chen-Fei;Chen, Chen;Zhou, Qin;Wang, Jin-Tao
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.14
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    • pp.5667-5672
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    • 2014
  • Folate receptor alpha ($FR{\alpha}$) mediates folate uptake by endocytosis, and while folate is essential to DNA methylation and synthesis and may have an important role in proliferating cells. $FR{\alpha}$ is known to be expressed in rapidly proliferating cells, including many cancer cell lines, but there has been no systematic assessment of expression in cervical cancer cell lines. The aim of the present study was to evaluate the effects of $FR{\alpha}$ on proliferation and apoptosis of cervical cells and correlation mechanism. In this study, we investigated the biological function of $FR{\alpha}$ in Hela cells using RNA interference. Cell proliferation was evaluated by Cell Counting Kit-8 (CCK8) assay, while cell cycling and apoptosis were assessed by flow cytometry, mRNA levels by real time-PCR and protein levels of $FR{\alpha}$, c-Fos and c-Jun by Western blotting. The results revealed that $FR{\alpha}$ was highly expressed in Hela cells and its silencing with a small interfering RNA (siRNA) inhibited cell proliferation and induced cell apoptosis, arresting the cell cycle in G0/G1 stages while decreasing the proportion in S and G2/M stages, and suppressed the expression levels of c-Fos and c-Jun. In conclusion, the results of this study indicated that $FR{\alpha}$ down-regulation might be capable of suppressing cervical cancer cell proliferation and promoting apoptosis. It suggested that $FR{\alpha}$ might be a novel therapeutic target for cervical cancer.

Development of Maintenance Simulation System and Prediction of Chloride Ion Permeation for Marine Concrete Structures (해양콘크리트 구조물의 염해 예측 및 유지보수 시뮬레이션시스템 개발)

  • Lee, Chang Su;Kim, Meyong Won
    • Journal of the Korea institute for structural maintenance and inspection
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    • v.17 no.1
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    • pp.64-75
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    • 2013
  • As both laboratory accelerated experiment and field exposure experiment were performed, at recent, the fifth field test at five year exposures was proceeded according to long period experimental plan. Field experiment, for the adoption of the developed evaluation model, which is consisted of the analysis of chloride penetration profile at gate bridges of sea-dike completed 30 years ago was carried out during upgrading the basic evaluation model with analyzing the annual field test data. The surface concentration of chlorides was replaced to the concentration of chloride of inner concrete near the surface chlorides among his research results at basic model. Maage's suggestion function was accepted too as a diffusion coefficient of chloride after verifying the change of diffusion coefficient by analysis of annual field test data. The comparison of field data with model predictions and the estimation of remaining life time demonstrates that the proposed updated model and maintenance simulation system can be used to predict the chloride penetration profile in the marine tidal zone and appropriate repair period and cost.

Taxonomic and Functional Changes of Bacterial Communities in the Rhizosphere of Kimchi Cabbage After Seed Bacterization with Proteus vulgaris JBLS202

  • Bhattacharyya, Dipto;Duta, Swarnalee;Yu, Sang-Mi;Jeong, Sang Chul;Lee, Yong Hoon
    • The Plant Pathology Journal
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    • v.34 no.4
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    • pp.286-296
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    • 2018
  • Maintenance of a beneficial microbial community, especially in the rhizosphere, is indispensable for plant growth and agricultural sustainability. In this sense, plant growth-promoting rhizobacteria (PGPR) have been extensively studied for their role in plant growth promotion and disease resistance. However, the impact of introducing PGPR strains into rhizosphere microbial communities is still underexplored. We previously found that the Proteus vulgaris JBLS202 strain (JBLS202) promoted growth of Kimchi cabbage and altered the relative abundance of total bacteria and Pseudomonas spp. in the treated rhizosphere. To extend these findings, we used pyrosequencing to analyze the changes in bacterial communities in the rhizosphere of Kimchi cabbage after introduction of JBLS202. The alterations were also evaluated by taxon-specific realtime PCR (qPCR). The pyrosequencing data revealed an increase in total bacteria abundance, including specific groups such as Proteobacteria, Acidobacteria, and Actinobacteria, in the treated rhizosphere. Time-course qPCR analysis confirmed the increase in the abundance of Acidobacteria, Actinobacteria, Alphaproteobacteria, and Betaproteobacteria. Furthermore, genes involved in nitrogen cycling were upregulated by JBLS202 treatment indicating changes in ecological function of the rhizosphere soil. Overall, these results indicate that introduction of JBLS202 alters both the composition and function of the rhizosphere bacterial community, which can have direct and indirect effects on plant growth. Therefore, we propose that long-term changes in bacterial composition and community-level function need to be considered for practical use of PGPRs.