Effects of Trichostatin A on TGF-B induced Prosate cancer cells

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Effects of Trichostatin A on TGF-B induced Prosate cancer cells.

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TSA. Trichostatin A (TSA) is an antifungal antibiotic isolated from Streptomyces that was first shown to be a potent inducer of cell differentiation and cell cycle arrest. inhibits the class I and II mammalian histone deacetylase. TSA inhibits the eukaryotic cell cycle during the beginning of the growth stage.

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Trichostatin A Prevents removal of acetyl groups Increased histone acetylation Altered gene expresssion.

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Pharmacological Properties of Trichostatin A, Focusing on the Anticancer Potential: A Comprehensive Review.

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TGF-B signalling pathway. CANONICAL very low frequency, high Of in HCC TGFß TGFß-R1/11 TGFß-R111 NONCANONICAL Cytokine TKR SMAD2 SMAD3 SMAD6/7 Growth factor GRB2 MEK ERK P13K mTOR TABI TAKI ROCK Cell cycle checkpoints Apoptosis Inflammation Cell cycle arrest Angiogenesis Proliferation Invasion.

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PROSTATE CANCER. Absolute numbers, Mortality, Males, in 2022 Prostate Continents Oceania 5 358 (1.3%) Northern America 39 605 (10.0%) Africa 55 744 (14.0%) LAC 61 056 (15.4%) Total . Cancer TODAY I IARC - https://gco.iarc.who.int/today Data version : Globocan 2022 C All Rights Reserved 2024 Asia 120 485 (30.3%) Europe 115 182 (29.0%) • 397 430 for Research on Cancer World Health Organization.

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Epithelial to Mesenchymal Transition. Loosening of intercellular junctions EMT Reduction of interactions with the ECM matrix, polarity loss Cytoskeletal reorganisation, Increased level of MMPs, increased migration basement membrane degradation, invasiveness Mesenchymal markers BASEMENT MEMBRANE Epithelial markers • E-cadherin Snail- Claudins Ocludins Cytokeratins Mucin-l 1 as an inductor • • N-cadherin Vimentin ß-catenin Fibronectin MMPs.

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OBJECTIVES. Determination of the Inhibitory Concentration 50 (IC50) of Trichostatin A (TSA) on Prostate Cancer Cells. Impact of Trichostatin A (TSA) on prostate cancer at both transcriptomic and proteomic levels. 3) Effects of Trichostatin A (TSA) on the migratory capabilities of prostate cancer cells.

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Seeding of PC3 Cells in 96 well plate (15,000 per well) Treatment with TSA (250, 500, 750, 1000, 1250) in nM DMSO MTT Read absorbance at 595 nm (Spectrophotometer).

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Seeding of PC3 cells in 6 well plate (3,00,000 per well) Starvation Treatment (6hr &24hr) (Control, DMSO, TGF-B,TSA, TGF-B+TSA) Protein Extraction Protein Estimation (Bradford assay).

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Seeding of PC3 cells in 6 well plates (3,00,000 per well) Starvation Treatment (6hr &24hr) (Control, DMSO, TGF-B,TSA, TGF-B+TSA) RNA Extraction cDNA synthesis qPCR.

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MTT. 0 250 500 750 1000 1250 1500 100 77.292576419213987 73.944687045123729 64.046579330422119 53.760310528869468 41.969917515769041 24.163027656477436.

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24 HOURS.

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Control DMSO TGF-B TSA TGF-b+TSA 1 1.1490610496950866 1.2064923770879801 0.99575713670789467 1.0137393710683962.

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SMAD7. SMAD7 Control DMSO TGF-β TSA TGF-β+TSA Actin.

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Vimentin. Vimentin Control DMSO TGF-β TSA TGF-β+TSA Actin.

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P-C-jun. Control DMSO TGF-B TSA TGF-b+TSA 1 0.97253347044060068 1.0622694960589341 0.69650992462208716 0.77523089501324471.

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N-cadherin. Control DMSO TGF-B TSA TGF-b+TSA 1 1.217650240970207 1.2988744169788846 0.64656237608807754 0.3563642882935259.

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E-cadherin. Control DMSO TGF-B TSA TGF-b+TSA 1 1.114562451142084 1.0732528596049846 0.96717367547143007 0.93559291285284718.

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pSMAD. Control DMSO TGF-B TSA TGF-b+TSA 1 1.0285949801060588 1.0694187717588997 0.95987467017630235 0.83084914388298614.

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sMAD7. Control DMSO TGF-B TSA TGF-b+TSA 1 0.8442274635525121 0.68989115707471327 0.63222435024222545 0.59155778228035527.

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Vimentin. Control DMSO TGF-B TSA TGF-b+TSA 1 0.71480109770765976 0.6657447242692347 0.52685340136137793 0.73508101087433342.

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P-C-Jun. Control DMSO TGF-B TSA TGF-b+TSA 1 1.0038968203322152 0.64997714218017433 1.0727096403087619 0.49959891269701506.

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N-cadherin. Control DMSO TGF-B TSA TGF-b+TSA 1 1.0792954550300486 1.2072945677903253 0.70714748087938617 0.3093192947317378.

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E-cadherin. Control DMSO TGF-B TSA TGF-b+TSA 1 0.83060836012153072 0.81540646002200634 0.38298286624353139 0.43280736702325412.