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ABSTRACT Background: Colorectal cancer is the third cause of cancer worldwide and a quarter of them are in the rectum. DEK oncogene is involved in several nuclear processes and can accelerate tumorigenesis. Objective: This study aims to evaluate the immunoexpression of DEK and Phospho-P38 proteins before neoadjuvant therapy in patients with rectum adenocarcinoma and correlate it with a clinical response and survival. Methods: Patients with adenocarcinoma of the middle and low rectum who underwent chemotherapy and radiotherapy followed by surgical tumor resection were included. The expression and quantification were studied by immunohistochemistry in the tumor biopsy tissues using a HScore system. Score ≥4 were considered positive and those with <4 negative. Results: 22 patients were included with a mean age of 63.55 years (SD: ±13.49). The clinical-stage before treatment was T3 on 72.7%, T4 on 18.2%, 31.8% were N1, 50% N0 and all M0. After chemo and radiotherapy, 54.6% were T3; 22.7% were classified as T2; 9.1% as T1, and 13.6% were T0. Among the tumors, 22.7% were positive for DEK and 63.6% positive for Phospho-P38. There was a positive correlation between DEK protein before treatment and pTNM stage (P=0.011). Phospho-P38 protein showed no correlation with these parameters. Patients with a negative HScore had a mean survival of 141.33 months (95%CI: 112.41-170.25) and those with a positive HSscore had a mean survival of 25.10 months (95%CI: 17.36-32.84; P<0.001). Conclusion: A higher expression of DEK was observed in advanced stages. Patients who presented DEK expression <4 had a higher survival, being a factor of worst prognosis.
RESUMO Contexto: O câncer colorretal é mundialmente, a terceira causa de câncer e um quarto destes estão localizados no reto. O oncogene DEK está envolvido em vários processos nucleares e pode acelerar a tumorigênese. Objetivo: Este estudo tem como objetivo avaliar a imunoexpressão das proteínas DEK e Fosfo-P38 antes da terapia neoadjuvante em pacientes com adenocarcinoma de reto e correlacioná-la com resposta clínica e sobrevida. Métodos: Foram incluídos pacientes com adenocarcinoma de reto médio e baixo submetidos à quimio e radioterapia seguida de ressecção cirúrgica do tumor. A expressão e quantificação foram estudadas por imuno-histoquímica nos tecidos de biópsia tumoral utilizando um sistema HScore. Escores ≥4 foram considerados positivos e aqueles com <4 negativos. Resultados: Foram incluídos 22 pacientes com média de idade de 63,55 anos (DP: ±13,49). O estágio clínico antes do tratamento era T3 em 72,7%, T4 em 18,2%, 31,8% eram N1, 50% N0 e todos M0. Após a quimio e radioterapia, 54,6% eram T3; 22,7% eram T2; 9,1% eram T1 e 13,6% T0. Entre os tumores, 22,7% foram positivos para DEK e 63,6% positivos para Phospho-P38. Houve uma correlação positiva para a imunoexpressão da proteína DEK e o estágio pTNM (P=0,011). A proteína fosfo-P38 não apresentou correlação com esses parâmetros. Pacientes com HScore negativo para DEK tiveram sobrevida média de 141,33 meses (IC95%: 112,41-170,25) e aqueles com HScore positivo tiveram sobrevida média de 25,10 meses (IC95%: 17,36-32,84) (P<0,001). Conclusão: Observou-se maior expressão de DEK em estágios avançados. Os pacientes que apresentaram expressão de DEK <4 tiveram maior sobrevida, sendo um fator de pior prognóstico.
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Objective:To investigate the dynamic change and clinical impact of DEK-NUP214 fusion gene in patients with acute myeloid leukemia (AML) receiving allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:Real-time quantitative polymerase chain reaction (RQ-PCR) and multicolor flow cytometry (FCM) were used to detect DEK-NUP214 gene expression and leukemia-associated immunophenotype (LAIP) in 15 newly diagnosed patients with positive DEK-NUP214 and receiving allo-HSCT from September 2012 to September 2017 at Peking University People′s Hospital. The clinical outcome was analyzed using Kaplan-Meier survival curves. The impact of DEK-NUP214 expression was analyzed by log-rank test.Results:The subjects were followed-up with a median period of 657 (62-2 212) days. The median DEK-NUP214 expression level at diagnosis was 488% (274%-1 692%). Thirteen patients achieved complete remission before allo-HSCT. Thirteen patients had a residual DEK-NUP214 expression of 0.38% (0.029%-738.9%) before allo-HSCT. After allo-HSCT, DEK-NUP214 expression in 9/13 patients remained positive, which dropped by around 500 folds (5.7-5 663.0 folds) within a month post-transplant. Five patients died and 2 patients relapsed. The 3-year cumulative incidence of relapse in patients with positive DEK-NUP214 before transplant was 17.5%±11.3% and the 3-year overall survival was 60.5%±13.8%. After allo-HSCT, DEK-NUP214-negative patients had a better outcome.Conclusion:Quantitative monitor of DEK-NUP214 fusion gene could be a sensitive indicator of MRD status after allo-HSCT.
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Objective:To investigate the clinical features and prognosis of patients with acute myeloid leukemia (AML) with DEK-CAN-positive.Methods:The clinical data of 6 AML patients with DEK-CAN-positive admitted to the First Affiliated Hospital of Zhengzhou University from August 2014 to January 2018 were retrospectively analyzed. Their clinical features, treatment and outcomes were summarized.Results:All 6 patients were female with a median age of 29 years (4-64 years). Among 6 cases, 5 cases were primary granulocytic leukemia partially differentiated (M 2) and 1 case was acute monocyte leukemia (M 5). There were 5 patients with elevated count of white blood cells in peripheral blood, and 1 patient had bone marrow dysplasia, and 2 patients had basophilia. Immunophenotypic analysis showed that all 6 patients were positive for CD34, CD13, CD38 and CD33. Fusion gene detection showed that DEK-CAN was positive and NPM1 mutations were negative in 6 patients; FLT3-ITD mutation was detected in 3 patients and WT1 mutation was detected in 2 patients. Chromosomal karyotyping showed that 2 cases had no split phase and 4 cases carried t(6;9). Follow-up details were available for 6 patients, only 1 patient achieved complete remission after the first course of induction chemotherapy, and the remaining 5 patients had no complete remission after the first course of chemotherapy, and died of complications in a short time. Conclusion:AML patients with DEK-CAN-positive have a very poor prognosis, low primary induced remission rate and high mortality.
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Aim To investigate the molecular mechanisms of the dual inhibitor of mammalian rapamycin target protein (mTOR) AZD8055 in migration and EMT process inhibition of the human cholangiocarcinoma cell line HuCCTl. Methods The viability of HuCCTl cells treated with different concentrations of AZD8055 was measured by MTT assay, and the colony formation ability of HuCCTl was detected by colony formation assay. The effect of AZD8055 on the motility of HuCCTl cells was examined by wound healing assay and Tran-swell assay. The expression levels of the protein associated with Akt/mTOR pathway, epithelial-mesenchymal transition (EMT) process and DEK were detected by Western blot. The interaction relationship between AZD8055, DEK and Akt signaling pathway was analyzed by STITCH and GeneMania databases. Cholangiocarcinoma cells'proliferation, migration capacities and Akt/mTOR signaling pathway-related protein expression levels were detected after DEK gene silencing. Results Compared with control group, AZD8055 inhibited the proliferation and migration capacities of cholangiocarcinoma cells, and suppressed the expression levels of Akt/mTOR signaling pathway-related markers, down-regulated DEK expression and inhibited EMT process. DEK silence significantly inhibited cell proliferation, migration and significantly decreased the phosphorylation levels of Akt, S6, and 4EBP1. Conclusions AZD8055 treatment inhibits the migration and EMT progression of HuCCTl cells, and its mechanism is associated with DEK down-regulation and inhibition of Akt/mTOR signaling pathway.
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Objective To study the influence of targeted silencing of DEK on the proliferation and cell cycle of human hepatoma cell lines.Methods The human hepatoma cells line HepG2 were routinely cuhured and the cells were divided into blank control group,siRNA control group and DEK siRNA group when the cells grew to 90％ tusion.The cells in blank control group were cultured normally without any treatment;the cells in siRNA control group and DEK siRNA group were transfected with siRNA expression vector and DEK siRNA expression vector mediated by LipofectamineTM2000 liposomes,respectively.The expression of DEK mRNA in HepG2 cells was detected by real-time polymerase chain reaction;the expression of DEK and CyclinD1 protein in HepG2 cells was detected by Western blot;the proliferation of HepG2 cells was detected by methyl thiazolyl tetrazolium method,and the cell cycle was observed by flow cytometry.Results The expression of DEK mRNA in the blank control group,siRNA control group and DEK siRNA group was 0.826 ±0.052,0.776 ±0.051 and 0.420 ±0.050 respectively;the expression of DEK protein in the blank control group,siRNA control group and DEK siRNA group was 0.691 ± 0.073,0.726±0.061 and 0.311 ±0.038 respectively;the expression of CyclinDl protein in the blank control group,siRNA cuntrol group and DEK siRNA group was 0.712 ± 0.069,0.780 ± 0.074 and 0.434 ± 0.039 respectively.The expressions of DEK mRNA,DEK protein and CyclinD1 protein in DEK siRNA group were significantly lower than those in the blank control group and siRNA control group (P ＜ 0.05);there was no statistic difference in the expression of DEK mRNA,DEK protein and CyclinD1 protein between the blank control group and siRNA control group(P ＜0.05).The proliferation ability of HepG2 cells in DEK siRNA group after transfection of 24,48,72,96,120 h was significantly lower than that in the blank control group and siRNA control group(P ＜0.05);there was no statistic difference in the proliferation ability of HepG2 cells between the blank control group and siRNA control group at each time point(P ＜ 0.05).The proportion of G0 + G1 phase cells in DEK siRNA group was significantly higher than that in the blank control group and siRNA control group(P ＜ 0.05);the proportions of S phase and G2 + M phase cells in DEK siRNA group were significantly lower than those in the blank control group and siRNA control group(P ＜ 0.05);there was no statistic difference in the proportion of G0 + G1 phase,S phase and G2 + M phase cells between the blank control group and siRNA control group (P ＜ 0.05).The result of Pearson correlation analysis showed that the expression of CyclinD1 protein was positively correlated with the expression of DEK mRNA and protein(r =0.909,0.899;P ＜ 0.05).Conclusion DEK siRNA can inhibit the proliferation of HepG2 cells,and change the cell cycle distribution through down regulating the expression of DEK gene in HepG2 cells.This process may be related to the down regulation of the expression of CyclinD1.
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Objective To detect the presence of DEK protein in the tissue and voided urine of patients with bladder urothelial carcinoma ,and an?alyze the relationship between DEK protein and clinical pathological classification of bladder cancer. Methods The expression of DEK protein was examined by immunohistochemistry in 48 bladder cancer tissues and 18 adjacent normal tissues(control group). The presence of DEK protein in voided urine was analyzed by western blot in 28 bladder cancer patients and 6 healthy individuals(control group). Results The positive ex?pression of DEK protein in bladder tumor tissues(73%)is higher than in adjacent normal tissues(33%,P<0.05). The expression of DEK in su?perficial bladder cancer(86%)was found higher than invasive bladder cancer(55%)with significant differences(P<0.05). The expression of DEK in urine samples of bladder cancer patients(0.834)was found higher than control group(0.242,P<0.05);Compared with the DEK expres?sion of control group,the sensitivity of diagnosis of bladder cancer with voided urine could be 82.1%. Conclusion DEK protein positive ex?pressed in tissues and voided urine of patients with bladder urothelial carcinoma. The expression was correlated with pathological classification of bladder cancer. The positive rate of DEK expression in early stage of tumor tissues is higher than late stage. The presence of DEK protein in tissues and voided urine can be considered as a suitable biomarker for bladder cancer potentially.
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DEK protein is an abundant chromatin protein in metazoans with highly conserved nuclear factor. This protein is a unique member of its family and is preferentially expressed in actively proliferating and malignant cells. Recently, much attention has been paid to the role of DEK protein in the development of various cancers, which was originally discovered in a subset of acute my-elogenous leukemia. Oncogene DEK is overexpressed in several malignancies including hepatocellular carcinoma, glioblastoma, retino-blastoma, bladder cancer, malignant melanoma, and cervical cancer. Oncogene DEK is a chromatin remodeling protein that supports cancer cell proliferation and invasion. In this review, we summarized research advancements in the correlation between DEK protein and oncogenesis.
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[ ABSTRACT] AIM:To investigate the effect of DEK downregulation on the apoptosis of gastric carcinoma SGC-7901 cells, and to explore its associations with NF-κB signaling pathway and apoptosis related proteins.METHODS:SGC-7901 cells with different treatments were divided into 3 groups including untreated group, control siRNA group and DEK siRNA group.The expression of DEK at mRNA and protein levels in the SGC-7901 cells was detected by real-time PCR and Western blot.The cell apoptosis was examined by flow cytometry.Furthermore, the activities of caspase-3 and caspase-9 in the SGC-7901 cells were investigated by Caspase-Glo?-3/9 kit.Finally, the expression of key regulatory pro-tein p65 of NF-κB signaling pathway and apoptosis-related proteins Bcl-2 and Bax in the SGC-7901 cells was investigated by Western blot.RESULTS:Compared with untreated group and control siRNA group, the expression of DEK at mRNA and protein levels was significantly downregulated in DEK siRNA group (P<0.05).In addition, the ratios of early phase apoptosis and total apoptosis in DEK siRNA group were markedly higher than those in untreated group and control siRNA group (P<0.05).Most notably, the decrease in p65 and Bcl-2 proteins, increase in Bax protein and the increases of caspase-3 and caspase-9 activities were observed in DEK siRNA group.CONCLUSION:Downregulation of DEK mediates cell apoptosis of gastric carcinoma may be tightly associated with NF-κB signaling pathway.
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Objective To construct the lentiviral vector of RNA interference(RNAi) for DEK,and to detect its effect on breast cancer cell growth.Methods The DEK siRNA was designed and constructed based on DEK sequence using a lentiviral vector.The lentivirul vector containing DEK siRNA was named PSIH-H1-DEK as confirmed by PCR and sequenceing.PSIH-H1-DEK was then packaged with accessory plasmids into lentivirus in 293T cells and selected for 2 weeks with puromycin ( puro ) before the mixed colonies stably expressing DEK siRNA were obtained and the DEK expression was detected by real time PCR( RT-PCR) and Western blotting.The effect of DEK siRNA on ZR75-1 cell growth was determined by cell counting kit.Results Western blot and RT-PCR showed that PSIH-H1-DEK siRNA could suppress DEK gene expression.Suppression of DEK could markedly inhibit the growth of ZR75-1 cells.Conclusion The lentivirus-mediated DEK siRNA is obtained,which will facilitate further research on DEK function in breast cancer development.
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Objective To investigate the pathological significance of DEK-AP-2α interaction in HER2 overexpres-sion and breast tumorigenesis. Methods The protein level of DEK,AP-2α and HER-2 in breast tumor tissues was detected by Western blot. The interaction of DEK and AP-2α in MDA-MB-453 mammary cancer cells was detected by immuno-coprecipitation. Furthermore the impact of DEK and AP-2α on HER2 expression was investigated by siRNA in MDA-MB-453 mammary cancer cells followed by semi-quantitive RT-PCR and Western blot. Results A correlation between DEK, AP-2α and HER2 levels in breast cancer tissues were found. The interaction between DEK and AP-2α in MDA-MB-453 cells was verified by co-immunoprecipitation assay. Depletion of DEK and AP-2α in MDA-MB-453 cell by siRNA cooperatively repressed HER2 expression at both the mRNA and protein levels. Conclusion Oncogene DEK has synergestic effect with AP-2α transcriptional factor to promote HER2 overexpres-sion in breast cancer.
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DEK protein's carboxy-terminal DNA-binding region(CBD)is a newly found DNA-binding domain of DEK,which contains several phosphorylation sites and has a close correlation with DEK protein's function in vivo and in vitro.Using prokaryotic expression system,the peptide of DEK protein's carboxy-terminal DNA-binding region(CDB)was expressed and purified.In detail,the CDB DNA fragment was constructed into pET-30a(+)vector,and E.coli BL21(DE3)competent cells were used as host cells.The fusion protein His-CBD was expressed by induction of IPTG and purified by Ni-NTA agarose.The result of SDS-PAGE showed that the molecular weight of the purified protein was about 10.7kDa.Electrophoretic mobility shift assay(EMSA)indicated that DEK-CDB prefered to bind to supercoiled form of DNA in vitro,it had similar character to the binding of whole length DEK protein with DNA.This suggested that the carboxy-terminal DNA-binding region of DEK protein might function on the binding of DEK protein to DNA partly.
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Among several newly identified oncogenes, dek and af4 are attractive targets for researchers interested with leukemia. In this study quantitative Real-Time RT-PCR technique was used to define alterations in expression of dek and af4 genes associated with acute promyelocytic leukaemia (APL) t (15; 17). RNA samples obtained from bone marrow aspirates of fourteen APL patients, cDNA portions were labelled with Syber Green 1 dye and LightCycler analysis have been performed. Expression changes in patients were found not significant in comparison to healthy donors for af4 (P=0.192) and dek (P= 0.0895). We suggest that af4 gene may have a role in leukomogenesis restricted to lymphoblastic lineage; also further studies must carry on with a larger series of patients in order to understand the relationship between the dek gene and APL. Our study was the first attempt for analysing dek and af4 genes in APL t (15; 17) patients by quantitative Real-Time RT-PCR. This rapid and sensitive method could be used to screen these genes in different types of leukaemia.