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Anti-Proliferative Activities of Vasicinone on Lung Carcinoma Cells Mediated via Activation of Both Mitochondria-Dependent and Independent Pathways

Biomolecules & therapeutics, v.26 no.4, 2018년, pp.409 - 416  

Dey, Tapan (Biological Sciences and Technology Division, CSIR-North East Institute of Science and Technology) ,  Dutta, Prachurjya (Biological Sciences and Technology Division, CSIR-North East Institute of Science and Technology) ,  Manna, Prasenjit (Biological Sciences and Technology Division, CSIR-North East Institute of Science and Technology) ,  Kalita, Jatin (Biological Sciences and Technology Division, CSIR-North East Institute of Science and Technology) ,  Boruah, Hari Prasanna Deka (Biological Sciences and Technology Division, CSIR-North East Institute of Science and Technology) ,  Buragohain, Alak Kumar (Centre for Biotechnology and Bioinformatics, Dibrugarh University) ,  Unni, Balagopalan (Biological Sciences, Assam Downtown University)

Abstract AI-Helper 아이콘AI-Helper

Vasicinone, a quinazoline alkaloid from Adhatoda vasica Nees. is well known for its bronchodilator activity. However its anti-proliferative activities is yet to be elucidated. Here-in we investigated the anti-proliferative effect of vasicinone and its underlying mechanism against A549 lung carcinoma...

주제어

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제안 방법

  • Whereas, Poly (ADP-ribose) polymerase (PARP) mediated programmed cell death is said to be caspase independent cell death pathway. In this study we examined both the pathways to evaluate the effect of VAS. The gene expression profile showed a few fold increases of FAS ligand (FasL), Fas, BAD, Cytochrome C, caspase-3, and PARP (Fig.
  • Therefore, antioxidant therapy may attenuate the ROS mediated cancer progression. The aim of the present study is to examine the anti-proliferative and antioxidant activities of vasicinone (VAS) in human lung epithelial cells and also to investigate the possible signaling pathways.

대상 데이터

  • The cell cultures were grown to form a monolayer (100% confluence) and then growth arrested for 24h in the absence of FBS before conducting experiments. Experiments used A549 cells of passage 68-78. The normal skin fibroblast cells were cultured in DMEM supplemented with 10% fetal bovine serum, penicillin (100 U/ml), streptomycin (100 μg/ml) and 22 mM sodium bicarbonate in a humidified atmosphere containing 5% (v/v) CO2.
  • The human alveolar epithelial cell line (A549) was obtained from the National Centre for Cell Sciences (Pune, India). The cells were cultured in Ham’s F-12K (Kaighn’s modification of Ham’s F-12 and Coon’s F-12 supplemented with higher concentrations of amino acids and pyruvate, as well as modified salts).
  • The human specific antibodies PARP (ab6079), Caspase 3 (ab966S), Bad (ab115311) and Bcl-2 (ab692) were purchased from Abcam, Inc (Cambridge, MA, USA). Vasicinone (CAS 486-64-6) was purchased from Cayman Chemicals, Michigan, USA. All other chemicals were purchased from Sigma (Saint Louis, USA) unless otherwise mentioned.

데이터처리

  • Data were analyzed statistically using one way analysis of variance (ANOVA) with Sigma Stat statistical software (Jandel Scientific, San Rafael, CA, USA). When data passed a normality test, all groups were compared using the Student–Newman–Keuls post hoc method.

이론/모형

  • Cells (100% confluency) were treated with respective concentrations for 72 h in a humidified incubator at 37°C and 5% CO2. Cell viability assay was performed using the MTT [3-(4,5-di-methylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide] method. The absorbance was read at a wavelength of 570 nm using micro-plate reader (BioTek Instruments, Inc, USA).
  • The experiments were carried out following the manufacturer’s protocol (Coral Clinical Systems, INDIA).
  • 2-Deoxyribose is oxidized by the hydroxyl radical generated by Fe3+ /Ascorbate/EDTA/H2O2 system (Fenton reaction) and degraded to malondialdehyde. The extent of deoxyribose degradation was measured by TBA method. The reaction mixture contained 2-deoxy-D-ribose (2.
  • The hydroxyl radical scavenging activity of VAS was examined by using the 2-deoxyribose oxidation method (Manna et al., 2010). 2-Deoxyribose is oxidized by the hydroxyl radical generated by Fe3+ /Ascorbate/EDTA/H2O2 system (Fenton reaction) and degraded to malondialdehyde.
  • , 2016). The present study evaluated the antiproliferative and antioxidant properties of VAS using A549 human lung cancer cell culture model.
  • When data passed a normality test, all groups were compared using the Student–Newman–Keuls post hoc method.
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