Expression and Significance of BLIMP-1 in Regulatory T Cells of Children with Aplastic Anemia / 中国实验血液学杂志

OBJECTIVE@#To study the expression of B lymphocyte-induced mature protein-1 (BLIMP-1) in regulatory T cells (Tregs) of children with aplastic anemia (AA), and analyze its correlation with the number of Tregs and the levels of inhibitory cytokines interleukin (IL)-10 and transforming growth factor (TGF)-β in plasma.@*METHODS@#The peripheral blood samples of 10 newly diagnosed AA children and 10 healthy children were collected for experiment. qPCR was used to detect FOXP3 and PRDM1 mRNA expression levels. Flow cytometry was used to detect the proportion of Tregs, the expression of BLIMP-1 in Tregs, and the levels of cytokines such as IL-2, IL-17A, IL-6, interferon (IFN)-γ, IL-10 and TGF-β in plasma. Pearson correlation model was used to evaluate the relationship between the expression of BLIMP-1 in Treg and the number of Tregs, as well as the levels of IL-10 and TGF-β in plasma.@*RESULTS@#Compared with control group, the proportion of Tregs in peripheral blood of AA children was decreased significantly (P<0.001); The plasma levels of proinflammatory cytokines IL-2, IL-6 and IFN-γ in AA children were increased significantly (P=0.033, P=0.031, P=0.006), and IL-17A also was increased but the difference was not statistically significant (P=0.052), while anti-inflammatory cytokines IL-10 and TGF-β were significantly reduced (P=0.048, P=0.002). The relative expressions level of FOXP3 and PRDM1 mRNA in AA children were significantly lower than those in control group (P=0.037, P=0.016). The expression of BLIMP-1 protein in Tregs of AA children was significantly lower than that in control group (P<0.001). The expression level of BLIMP-1 protein in Tregs was positively correlated with the percentage of Tregs in lymphocytes (r=0.671, P=0.001), and was also positively correlated with the levels of IL-10 and TGF-β in plasma (r=0.500, P=0.029; r=0.486, P=0.030).@*CONCLUSION@#The expression of BLIMP-1 in Tregs of AA children is impaired, and the low expression of BLIMP-1 is related to the decrease of the number in Tregs and IL-10 and TGF-β expressions.

Correlation of Blimp1 with ATF4/CHOP Signaling Pathway in Multiple Myeloma U266 Cells / 中国实验血液学杂志

OBJECTIVE@#To investigate whether Blimp1 plays an anti-apoptosis role in myeloma by interfering with ATF4/CHOP cell apoptosis pathway induced by endoplasmic reticulum stress, and to explore the anti-myeloma mechanism of aspirin.@*METHODS@#The bone marrow fluid of 40 newly diagnosed multiple myeloma patients without treatment and 30 control people with relatively normal bone marrow was collected. Flow cytometry was used to separated the normal and abnormal plasma cells, LV-Blimp1-RNAi (40051-2) recombinant lentivirus down-regulates the expression of Blimp-1 in U266 cell line and detected the changes of the expression of ATF4 and CHOP gene. U266 cells were stimulated by aspirin at different concentrations (0, 0.5, 2.5, 5.0 mmol/L) in vitro. Then the effect of aspirin on proliferation of U266 cells was measured by CCK-8 assay, the mRNA expression levels of Blimp1, ATF4 and CHOP in four groups were detected by real-time PCR.@*RESULTS@#The expression level of Blimp1 in phenotype abnormal plasma cells was significantly increased as compared with normal cells, while the expression of ATF4 and CHOP in phenotype abnormal plasma cells was significantly decreased as compared with normal cells (P＜0.05). In the case of MOI=100, the transfection efficiency of U266 cells was beyond 80% as detected by fluorescence microscopy. Compared with blank conrol and negatine control groups, Blimp1 mRNA expression level in positive group was significantly reduced while ATF4 and CHOP expression significantly increased. CCK-8 showed that the proliferation activity of U266 cells could be inhibited by aspirin, which showed a time-and dose-dependent manner; at the same time, the expression level of Blimp1 in U266 cells were decreased with the increasing of aspirin concentration, while the expression level of ATF4 and CHOP was increased with the increasing of aspirin concentration.@*CONCLUSIONS@#Blimp1 may display the anti-apoptosis of myeloma cells through interfering with ATF4/CHOP signaling pathway; low dose of aspirin may play anti-myeloma effect by inhibiting the expression of Blimp1 in myeloma cells.

PRDM1 expression and its relationship with PI3K/AKT pathway activation in extranodal NK/T cell lymphoma-nasal type / 中华血液学杂志

Objective: To investigate the expression of PRDM1 and its relationship with PI3K/AKT pathway activation in extranodal NK/T cell lymphoma-nasal type. Methods: Immunocytochemistry and Western blot were used to detect the expression of PRDM1 and p-AKT in 10 EN-NK/T-NT tissue or 3 cell lines (PRDM1-positive YT cell line, PRDM1-negative NKL and NK92 cell lines). Nanostring gene expression profiling technique was used to detect the activation of the PI3K/AKT pathway in normal nasal mucosa, PRDM1-negative and positive EN-NK/T-NT tissue. MTS was used to detect cell proliferation, and flow cytometry was used to detect cell cycle and apoptosis. Results: Nanostring gene expression profiling revealed that genes associated with PI3K/AKT signaling pathway (eg, IL-7, BRCA1, ITGA8, IL2RB, FASLG, CDK2, COL27A1, CSF3R, KITLG and IL-6) were highly expressed in EN-NK/T-NT cases (P<0.05). Also, we found that p-AKT was highly expressed in YT cell line, but lower or not expressed in NK92 and NKL cells. In addition, LY294002, a PI3K/AKT pathway inhibitor, increased PRDM1 and PTEN expression in a dose dependent manner in YT cells. More importantly, YT cell were treated with 20 μmol/L LY294002 48 h, the proliferation rate was significantly decreasing (58.18% vs 100.00%, t=12.770, P=0.006), and the proportion of cells in G(1) phase was significantly increased (30.05% vs 76.93%, t=11.570, P<0.001). However, there was no significant difference in cell proliferation and cell cycle between NKL cells and control group (P>0.05). Conclusion: The activation of PI3K/AKT pathway is positive associated with the expression of PRDM1 in EN-NK/T-NT, and inhibition of PI3K/AKT pathway may have significant therapeutic potential for PRDM1-positive EN-NK/T-NT.

Análise da forma fosforilada do fator de choque térmico 1 (pHSF 1) em carcinomas de células escamosas de língua oral / Analysis of the phosphorylated form of heat shock factor 1 (pHSF 1) in oral tongue squamous cell carcinomas

O carcinoma de células escamosas de língua oral (CCELO) apresenta altas taxas de morbimortalidade, apesar dos avanços recentes no tratamento das neoplasias. Assim, várias pesquisas vêm tentando detectar alterações morfológicas ou identificar biomarcadores que possam apresentar poder preditivo de recidiva e metástase e novas estratégias e/ou opções terapêuticas mais individualizadas com intuito de melhorar o prognóstico destes pacientes. O fator do choque térmico 1 (HSF1) atua de diferentes formas na progressão tumoral, favorecendo o surgimento, desenvolvimento e invasividade tumoral e sua superexpressão vem sendo pesquisada em neoplasias. Esta pesquisa objetivou analisar se a forma fosforilada do fator de choque térmico 1 (p-HSF1) exerce alguma influência na patogênese do CCELO. Foi realizado um estudo imunoistoquímico em 69 casos de CCELO, verificando-se a expressão do p-HSF1 e correlacionado esta imunoexpressão com alguns parâmetros clinicopatológicos e sobrevida dos pacientes. Foi avaliado e comparado o escore de imunopositividade do p-HSF1 entre CCELO e mucosa oral normal (MON) e esta expressão foi ainda correlacionada aos sistemas de gradação histológica propostos por Brandwein-Gensler et al. (2005) e pelo modelo BD (ALMANGUSH et al., 2014). As curvas de associação de análise de sobrevida aos outros parâmetros foram realizadas pelo método Kaplan Meier e as diferenças dessas curvas submetidas ao teste log-rank (p<0,05). Verificou-se maior escore de imunoexpressão (p<0,001) de p-HSF1 em CCELOs em relação a MON, sugerindo que esse fator pode influenciar a patogênese destes tumores. A imunoexpressão do p-HSF1 não foi associada aos parâmetros clinicopatológicos pesquisados nesta amostra. O tamanho do tumor (T) T3/T4 foi associado ao alto risco tanto pelo sistema de Brandwein-Gensler et al. (2005) quanto pelo modelo BD (ALMANGUSH et al., 2014), sugerindo que tumores maiores podem ser associados a piores parâmetros histopatológicos. Os resultados da análise por meio do método BD, mostraram relevância prognóstica, uma vez que pacientes classificados como de alto risco por este sistema, foram associados a uma menor sobrevida global e maior número de óbitos, sugerindo sua possível inclusão como uma ferramenta útil na determinação do prognóstico de pacientes com CCELO (AU).

The squamous cell carcinoma (SCC) of the oral tongue presents morbimortality high levels although recent achievements of malignancies treatment. This way, a lot of researches are trying to detect morphological alterations or identify biomarkers that may present recurrence prediction power and metastasis and new strategies and/or more individualized therapeutic options in order to improve the prognosis of these patients. The Heat shock factor 1 (HSF1) acts in different ways of tumoral progressing, facilitating the appearance, development and tumoral invasiveness and its overexpression has been researched in malignancies. This research had the aim to analize if the phosphorylated form of Heat shock factor 1 (p-HSF1) carries some influence in the SCC of the oral tongue pathogenesis. There was an immunohistochemical study in 69 cases of oral tongue squamous cell carcinoma observing the p-HSF1 expression and correlating this immunoexpression with some clinicopathological parameters and patients' survival. It was evaluated and compared the immunohistochemical score of the p-HSF1 between the squamous cell carcinoma (SCC) of the oral tongue and normal oral mucosa (MON) and this expression was correlated to the histological gradation systems proposed by Brandwein-Gensler et al. (2005) and by the model BD (ALMANGUSH et al., 2014). Survival analysis of association curves with the other parameters were carried out with the Kaplan Meier method and the differences of these curves submitted to the test logrank (p<0,05). It was found a bigger immunoexpression score (p<0,001) of p-HSF1 in squamous cell carcinoma (SCC) of the oral tongue in relation to the MON, suggesting this factor may influence the tumors pathogenesis.. The tumor size ( T ) T3/ T4 was associated to the high risk not only by system of Brandwein-Gensler et al. (2005) as by model BD (ALMANGUSH et al., 2014), suggesting bigger tumors may be associated to worse histopathological parameters. The analysis result through the BD method, showed prognostic implication, since as patients classified as high risk by this system were associated to a smaller global survival and bigger death number, suggesting its possible inclusion as a useful tool in the prognosis determination of patients with squamous cell carcinoma (SCC) of the oral tongue (AU).

Alterations of CD4+CXCR5+Tfh cells and its transcription regulatory factors in children with asthma / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the alterations of follicular T helper cells (CD4(+)CXCR5(+)Tfh cells, Tfh) on circulating T lymphocytes in children with asthma, and to study the expression of transcription regulatory factors BCL-6 and BLIMP-1 mRNA.</p><p><b>METHODS</b>Sixty-four children with asthma and 25 healthy controls were enrolled in this study. On the basis of the disease, the children with asthma were classified into acute phase group (n=36) and remission phase group (n=28). The flow cytometry was used to detect the proportion of CD4(+)CXCR5(+)Tfh cells on CD4(+)T lymphocytes. Real-time PCR was performed to detect the levels of BCL-6 mRNA and BLIMP-1 mRNA. The double -antibody Sandwich ELISA was used to detect plasma concentrations of total IgE, IL-2, IL-6 and IL-21.</p><p><b>RESULTS</b>The proportion of CD4(+)CXCR5(+)Tfh cells was significantly higher in the acute group than in the control group and the remission group (P<0.05). Transcription levels of BCL-6 mRNA were significantly higher, while the inhibitory factors BLIMP-1 mRNA was significantly lower in the acute group than in the remission group and control group (P<0.05). The plasma concentration of IL-6 in the acute group increased significantly compared with the control group (P<0.05). Plasma concentrations of total IgE and IL-21 increased significantly, in contrast, plasma IL-2 concentration decreased significantly in the acute group, compared with the control group and the remission group (P<0.05). Correlation analysis showed that both IL-21 and IL-6 concentrations were positively correlated with the proportion of CD4(+)CXCR5(+)Tfh cells (r=0.76, r=0.46 respectively; P<0.05), while IL-2 level was negatively correlated with the proportion of Tfh cells (r=-0.68, P<0.05).</p><p><b>CONCLUSIONS</b>The abnormal proportion of CD4(+)CXCR5(+)Tfh cells might be involved in the immunological pathogenesis of acute asthma in children. The increased expression of BCL-6 mRNA and decreased expression of BLIMP-1 mRNA as well as the alterations of plasma total IgE, cytokines IL-2, IL-6 and IL-21 in microenvironment might be account for the increased proportion of CD4(+)CXCR5(+)Tfh cells in children with acute asthma.</p>

Research progress on B lymphocyte-induced maturation protein 1 and its relationship with the development of lymphoma / 中国实验血液学杂志

Many studies show that as a transcription factor, B lymphocyte-induced maturation protein 1 (Blimp 1) is the master regulator of plasma-cell differentiation. The abnormality of Blimp 1 plays an important part in the genesis and development of lymphoma. This review introduces and summarizes Blimp 1's protein structure and functions, its role in B cell differentiation, its main target genes and the mechanism of its transcriptional repressor activity. Besides, the relationship between Blimp 1 gene mutation or Blimp 1 protein expression reduction and the development of DLBCL is preliminary summaried.

Analysis of the immunohistochemical subtypes and prognosis of primary diffuse large B cell lymphoma of the central nervous system / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To analyze the subtypes of primary diffuse large B cell lymphoma of the central nervous system (CNS DLBCL) and to explore the relationship between the subtype classification and prognosis.</p><p><b>METHODS</b>Immunohistochemical staining was used to determine the expression of CD20, CD3, CD10, Bcl-6, Mum-1, CD5, Bcl-2, Ki-67, FOXP-1, GCET-1, BLIMP-1 and LMO-2 antigens on paraffin-embedded sections of 47 cases. Hans, Choi and Tally subtypes were classified, and univariate and multivariate analyses were used to elucidate the relationship between the subtypes and prognosis.</p><p><b>RESULTS</b>In the 47 cases, the expression of Bcl-2 in the tumor cells was 46.8%, CD10 4.3%, Bcl-6 70.2%, Mum-1 53.2%, GCET-1 36.2%, BLIMP-1 4.3%, FOXP-1 63.8% and LMO-2 19.2%. The positive rate of Ki-67 was 30% to 95%, with a median of 80%, of which 12 cases (25.5%) was > or = 90%. The Hans subtype classification showed 16 cases (34.0%) were of GCB type and 31 cases (66.0%) of non-GCB type. The Choi subtype classification showed 16 cases (34.0%) were of GCB type and 31 cases (66.0%) of ABC type. The Tally subtype classification showed 6 cases (12.8%) were of GCB type and 41 cases (87.2%) of non-GCB type.</p><p><b>CONCLUSIONS</b>The results of this study show that there is no significant correlation between the three subtypes and prognosis. The prognosis is correated with post-operative radiotherapy, chemotherapy and MTX therapy.</p>

Rs548234 polymorphism at PRDM1-ATG5 region susceptible to rheumatoid arthritis in Caucasians is not associated with rheumatoid arthritis in Chinese Han population / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>A previous study has shown that rs548234 polymorphism at PRDM1-ATG5 region is associated with rheumatoid arthritis (RA) in Caucasian populations. The aim of this study was to investigate the effect of rs548234 polymorphism at PRDM1-ATG5 region on susceptibility to RA in Chinese Han population.</p><p><b>METHODS</b>We genotyped 848 RA patients and 1431 matched healthy controls for rs548234 single-nucleotide polymorphism (SNP) with a predesigned TaqMan SNP genotyping assay. Association analyses were performed on the whole data set and on rheumatoid factors (RF) and anti-cyclic citrullinated peptides (anti-CCP) antibody. Finally, we carried out combined analysis of rs548234 association with RA based on the published data.</p><p><b>RESULTS</b>No significant difference in the genotype distribution between RA patients and healthy controls for rs548234 (C/T) polymorphism was found in Chinese Han population, neither in whole data set nor in stratified subsets, e.g. RF and anti-CCP status. Association analysis in different ethnic groups showed that rs548234 at PRDM1-ATG5 region was associated with RA in Caucasian ancestry but not in East Asian population.</p><p><b>CONCLUSIONS</b>Our results showed no involvement of rs548234 at PRDM1-ATG5 region in the susceptibility or clinical relevance of RA in Chinese Han population.</p>

Blimp-1 protein and Hans classification on prognosis of diffuse large B-cell lymphoma and their interrelation / 癌症

<p><b>BACKGROUND AND OBJECTIVE</b>Diffuse large B-cell lymphoma (DLBCL), the most common type of non-Hodgkin's lymphoma (NHL), is heterogeneous on molecular and clinical levels, therefore, its prognosis is difficult to predict. This study aimed to evaluate the value of Blimp-1 protein and Hans classification in predicting the prognosis of DLBCL and their interrelation.</p><p><b>METHODS</b>The clinical records of 136 patients with DLBCL were reviewed. The patients were followed up for 5-80 months (median, 39 months). Immunohistochemical staining for CD10, MUM1, Bcl-6, and Blimp-1 were performed on paraffin-embedded tumor tissues from the 136 patients. The correlations of Blimp-1 protein and Hans classification in prognosis of DLBCL and their interrelation were analyzed.</p><p><b>RESULTS</b>Blimp-1 was detected in 38 (30.0%) patients, and was associated with a significantly shorter overall survival (OS) (P = 0.030). Using the Hans classification based upon the expression of CD10, Bcl-6, and MUM1, 54 patients had germinal center B-cell (GCB) phenotype and 82 had non-GCB phenotype. The 5-year OS rate was 75% in the GCB group and 52% in the non-GCB group (P = 0.020). The positive rate of Blimp-1 was 22.2% in the GCB group and 31.7% in the non-GCB group (P = 0.329). The Cox regression multivariate analysis showed that international prognosis index (IPI) and Hans classification had independent prognostic significance, whereas Blimp-1 was not an independent prognostic factor.</p><p><b>CONCLUSIONS</b>The patients with GCB subtype of DLBCL had better prognosis than the non-GCB subtype. High level of Blimp-1 expression in the patients with DLBCL implies a shorter survival, but it is not associated with Hans classification.</p>

Effect of low-dose 2-methoxyestradiol on the expressions of two isoforms of PRDM1 gene in myeloma / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the effects of low-dose 2-methoxyestradiol (2ME2) on the expression of two isoforms of PRDM1 gene (PRDM1alpha with and PRDM1beta without PR-domain) during the myeloma cell differentiation.</p><p><b>METHODS</b>In myeloma cell line NCI-H929, RPMI8226, KM3 and LP-1, PRDMla and PRDM1beta transcripts were detected by real-time quantitative PCR after treatment with low-dose of 2ME2 (0.5 micromol/L) for 0 h, 48 h and 72 h.</p><p><b>RESULTS</b>Both PRDM1alpha and PRDMIbeta3 were time-dependently upregulated, and the PRDMlalpha/beta ratio was elevated from 1.461 +/- 0.033 to 2.663 +/- 0.381 (P < 0.01), 1.929 +/- 0.334 to 2.727 +/- 0.362 (P < 0.05), 1.471 +/- 0.012 to 4.367 +/- 0.243 (P < 0.01) and 1.660 +/- 0.042 to 3.059 +/- 0.167 (P < 0.01) in NCI-H929, RPMI8226, KM3 and LP-1 cells respectively.</p><p><b>CONCLUSION</b>Low dose of 2ME2 can induce differentiation of myeloma cells as well as up-regulate the expression of PRDMlalpha and PRDM1beta in these cells. Elevation of PRDM1alpha/beta ratio was in a time-dependent manner and positively associated with cell differentiation and in accordance with Ying-Yang mechanism of PR-domain-containing gene family.</p>

Regulation of c-myc expression by Blimp-1 during the differentiation of myeloma cells / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the repression of c-myc induced by Blimp-1 in 2-methoxyestradiol(2ME2)-mediated differentiation of multiple myeloma cells.</p><p><b>METHODS</b>CZ-1 and LP-1 myeloma cells lines were exposed to 0.5 micromol/L of 2ME2 and 0.5 micromol/L of 2ME2 + antisense oligonucleotides (ASODN) for 72 h. The effects on transcription of c-myc mRNA were studied by RT-PCR. The c-myc protein was assayed with Western blot. The changes of the cell lines in morphology, expression of surface CD49e and quantity of immunoglobin light chain secretion in the supernatant were studied.</p><p><b>RESULTS</b>Incubation of the cells with 0.5 micromol/L of 2ME2 could up-regulated Blimp-1 expression and increase the expression of c-myc gene. In contrast, c-myc expression was decreased with Blimp-1 expression down-regulated by ASODN and the cell lines differentiation was arrested.</p><p><b>CONCLUSION</b>Blimp-1 could directly repress the expression of c-myc in 2ME2-mediated differentiation induction of multiple myeloma cells.</p>

Expression of tumor suppressor gene PRDM1 in lung cancers / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To determine the expression of tumor suppressor gene PRDM1 in lung cancers.</p><p><b>METHODS</b>Forty-five cases were enrolled in this study, including squamous cell carcinoma (20 cases), adenocarcinoma (15 cases), and small cell cancer (10 cases). PRDM1 protein was detected in paraffin-embedded tissue by immunohistochemistry. Tumor cells in lung cancers were further selected by laser microdissection for RT-PCR analysis. PRDM1 protein in frozen tissue was also detected by Western blot.</p><p><b>RESULTS</b>(1) PRDM1 protein was found in paraffin-embedded tissues in 90.0% (18/20) of squamous cell carcinoma, 13.3% (2/15) of adenocarcinoma, and 0 (0/10) small cell lung cancer. Squamous cell carcinoma predominantly expressed PRDM1 protein ( P < 0.01). (2) Gene product of PRDM1 DNA binding region was not found in microdissected tumor cells, but an abnormal PRDM1 protein about 70 KD was detected simultaneously in whole tumor tissue.</p><p><b>CONCLUSION</b>PRDM1 may be considered as a specific biomarker in pulmonary squamous cell carcinoma. The abnormal PRDM1 expression both at transcriptional and protein levels indicated that this tumor suppressor gene lost its function, which may become a new target in the strategy of treatment for lung cancers.</p>