| Background:Acute lymphoblastic leukemia(ALL)is the most common type of blood or bone marrow(BM)cancer among young children,although adults can get it as well.Drug resistance remains a significant cause of treatment failure.There is an increasing demand for developing a new therapeutic approach for the treatment of an acute form of lymphoblastic leukemia.FKBP51 is an immunophilin which is physiologically expressed in lymphocytes and is involved in sustaining cell growth,malignancy,and resistance to therapeutics.Objective:To investigate the effect of FKBP51 protein to elucidate its role in acute lymphocytic leukemia using Jurkat cell line.The study hypothesizes that FKBP51 is overexpressed in acute lymphocytic leukemia such as can be used for chemosensitivity and as a biomarker for tumor development/progression and response to anticancer drugs.Methods:Jurkat cell line treated with lentivirus vectors human immunodeficiency virus-1(HIV-1)for FKBP51 overexpression in acute lymphocytic leukemia was detected by western blot.The cell proliferation in Jurkat cells of FKBP51and FKBP51-shRNA knockdown was transfected with lentivirus assessed by cell counting Kit-8.For FKBP51-shRNA knockdown gene sequence amplification qPCR was used while cell cycles were determined by Muse(?)cell analyzer Kit,as well as mechanism a serine/threonine protein kinase(AKT),and nuclear factor-kappa B(NF-κB)signaling pathway.The statistical data were assessed by two-tailed students-test,one-way analysis of variance(ANOVA),and two-way independent variables among the group,analyzed by Graph Pad Prism 6 software,and statistical package for the social sciences(SPSS)version 25.The data are shown as the mean ± standard deviation(SD);P<0.05 was considered to be significantResults:Our results revealed that overexpression of FKBP51 enhances Jurkat cell proliferation,and its silencing inhibit Jurkat cell proliferation.In addition cell cycle demonstrated that overexpression decreased G0/G1 phase and increased G2/M and S-G2/M phase in number.Whereas FKBP51-shRNA increased G0/G1 phase cells and decreased G2/M phase cell numbers.Western blot analysis revealed that overexpression decreased phosphorylation activity of AKT,while there was a significant increase in the phosphorylation activity in an FKBP51 knockdown.The level of NF-KBp65,both in cytoplasm and nucleus,was found to be higher in FKBP51 overexpression cells than that in the control in a quiescent state for relatively short time 5 minutes and reversed atl5mintutes following TNFα stimulating.However,the level of NF-κBp65,pIKKα,and IκB remained still significantly higher than that in control.Similarly,the impaired NF-κB p65 and pIKKα activity in Jurkat cells depleted of FKBP51 gradually increase within 5 minutes in response to TNFα stimulus.Overexpression of FKBP51 increased Jurkat resistance to Dexamethasone(Dex),and promoted Jurkat cell proliferation in dose and time-depended manner.While FKBP51 gene knockdown sensitizes Jurkat cell lines to Dex and decreases cell proliferation.Our study found that higher dose of Dex 8,16,and 32μM decrease cell proliferation after 72 hours.Conclusions:Overexpression of FKBP51 protein promotes acute lymphocytic leukemia by increasing cell proliferation,specifically T-cell lines,possibly involved downstream of the AKT,suggesting that FKBP51 may play a significant role in the anti-apoptotic pathway mediated by NF-κB.Therefore,it will be interesting to consider FKBP51 protein as a future biomarker for tumor development and chemoresistance in clinical medicine... |
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