Protein arginine methyltransferases, which proceed the post-translational modification of both histones and non-histones, play an important role in many biological pathways. Protein arginine methyltransferase 5 (PRMT5) is a major enzyme responsible for symmetric di-methylation of arginine residues and has been suggested as a potential therapeutic target for tumors.In the past decade,the discovery and development of PRMT5 inhibitors have become one of the most important research fields.This article introduces the structure and biochemical function of PRMT5 and its correlation with cancer reviews, the binding modes and biological data of PRMT5 inhibitors under development, and discusses the clinical application potential of PRMT5 inhibitors in the treatment of cancer.

This study was aimed to quantitatively detect the expression levels of gfi-1 gene in leukemia patients, and to investigate the effect of gfi-1 gene silenced by short hairpin RNA (shRNA) on proliferation of leukemia cell line K562. Quantitative real-time PCR and Western blot were used to detect the mRNA and protein expression levels of GFI-1 in newly diagnosed patients with leukemia. One pair of oligonucleotide sequences targeted at human gfi-1 mRNA were designed and synthesized. The annealed oligonucleotide fragments were subcloned into pLVTHM vector. Virus particles were collected when the control and shRNA vectors had been co-transfected with the psPAX2 packaging plasmid and the envelope plasmid pMD2 G into HEK-293T cells using Lipofectamine 2000. The K562 cells were transfused with 1 x 10⁶ recombinant lentivirus-transfusing units plus 6 microg/ml of polybrene. Rea-time PCR and Western blot were used to detect the expressions of gfi-1 and bax mRNA after lentivirus transfusion. CCK-8 assays was used to evaluate the proliferation potential of cells. The results showed that the gfi-1 expression level in all leukemia patients was significantly higher than that in normal group (p < 0.05); the gfi-1 mRNA expression in chronic myeloid leukemia (CML) and acute myeloid leukemia (AML) or acute lymphoid leukemia (ALL) patients was significantly higher than that in normal group (p < 0.05); but the difference of gfi-1 mRNA expression between AL and CML or ALL and AML was not significant. Notably, the gfi-1 mRNA expression level had a positive correlation with high white blood cell count of > 20.0 x 10⁹/L (p < 0.05). As was expected, the mRNA and protein level of gfi-1 was reduced significantly in K562 cells after lentivirus transfusion, whereas the mRNA and protein level of bax was upregulated. And CCK-8 assay showed that gfi-1 gene silencing can inhibit K562 proliferation. It is concluded that gfi-1 expression is upregulated in leukemia patients and may contribute to leukemogenesis. The gfi-1 specific shRNA mediated by lentivirus can effectively down-regulate the expression of gfi-1 and inhibit the proliferation of K562 cells, which lay a basis for further research on gene therapy in leukemia.
Adolescent ; Adult ; Aged ; Case-Control Studies ; Cell Proliferation ; DNA-Binding Proteins ; genetics ; Female ; Gene Silencing ; Genetic Vectors ; Humans ; K562 Cells ; Lentivirus ; genetics ; Leukemia ; genetics ; pathology ; Male ; Middle Aged ; Plasmids ; RNA, Small Interfering ; genetics ; Transcription Factors ; genetics ; Young Adult

This study was aimed to explore the synergistic effect of 2-methoxyestradiol (2-ME2) and bortezomib (Bor) on the proliferative inhibition and apoptosis of U266 cell line and its possible mechanism. The cells were treated with 2-ME2, Bor alone and 2-ME2 combined with Bor, respectively. The cell viability and proliferative curve were detected by CCK8, the cell apoptosis was detected by caspase 3/7 activity test, cell cycle status was analyzed by flow cytometry, and real-time PCR was used to detect the mRNA expression of P21, BAX and BCL-2. The results showed that compared with cells treated with 2-ME2 or Bor alone, the proliferative potential of cells in combination group was significantly inhibited (p < 0.05), and apoptosis rate markedly increased (p < 0.05), cell cycle was arrested at G(1)-S phase, the mRNA expressive level of P21 and BAX increased, while the expression of BCL-2 decreased. It is concluded that 2-ME2 combined with Bor synergistically inhibits cell proliferation and induces apoptosis in U266 cell line. The possible mechanism may be associated with its effect of up-regulating P21 and BAX expressions.
Apoptosis ; drug effects ; Boronic Acids ; pharmacology ; Bortezomib ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; drug effects ; Drug Synergism ; Estradiol ; analogs & derivatives ; pharmacology ; Humans ; Pyrazines ; pharmacology

The objective of this study was to explore the effect of bortezomib (BZM) on lymphoma cell line CA46 and its relative mechanisms in vitro. The effects of BZM on the proliferation and apoptosis of CA46 cells were assayed by MTT method and flow cytometry respectively. The effect of BZM on the expression levels of procaspase-3 and BCL-2 protein were detected by Western blot. The results indicated that the BZM could inhibit the growth of CA46 cells significantly and the concentration of 50% growth inhibition (IC₅₀) at 24 and 48 hours were 53.19 and 19.68 nmol/L respectively. After treatment with 20, 40, 80 nmol/L BZM for 24, 48 and 72 hours, a dose- and time-dependent apoptosis of CA46 cells could be observed. After treatment with 20 nmol/L BZM at different time point, a time-dependent reduction of procaspase-3 and BCL-2 protein expression in CA46 cells was found. It is concluded that the BZM can inhibit the proliferation and induce the apoptosis of CA46 cells, which relative mechanism may involve the reduction of BCL-2 and the activation of caspase 3.
Apoptosis ; drug effects ; Boronic Acids ; pharmacology ; Bortezomib ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphoma ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Pyrazines ; pharmacology

Adult ; Central Nervous System Diseases ; diagnosis ; surgery ; Hamartoma ; diagnosis ; surgery ; Humans ; Male

This study was aimed to explore the expression level and correlation of MCL-1 and miR-29a in extranodal NK/T-cell lymphoma (ENKTCL) tissue. Maxvision immunohistochemistry technique and real time fluorescent quantitative PCR were used to detect the expression level of MCL-1 and miR-29a in tissue of 20 patients with ENKTCL and 10 patients with proliferative lymphadenitis, respectively. The results showed that the expression of MCL-1 protein were higher in patients with ENKTCL than that in patients with proliferative lymphadenitis, but there were no significant correlation between MCL-1 overexpression and age, sex, Ann Arbor stage and International Prognostic Index (IPI), respectively. Correlation analysis indicated that there was significant negative correlation between miR-29a expression and MCL-1 expression (r = -0.59, P = 0.016). It is concluded that miR-29a may target MCL-1 gene, regulate its expression, then participate in tumorigenesis and development of ENKTCL.
Adolescent ; Adult ; Aged ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphoma, Extranodal NK-T-Cell ; genetics ; pathology ; Male ; MicroRNAs ; genetics ; Middle Aged ; Myeloid Cell Leukemia Sequence 1 Protein ; genetics ; Young Adult

This study was aimed to establish a high-throughput luciferase reporter system, through which to screen and identify miRNAs directly targeting p21, and to explore the biological function and significance of these miRNAs. Molecular cloning technique was used to construct and identify two lentivirus-expressing vectors-pWPXL-Luc and pWPXL-Luc-P21-3'UTR, virus particles were collected after the pWPXL-Luc or pWPXL-Luc-P21-3'UTR vectors were co-transfected with the psPAX2 packaging plasmid and the envelope plasmid pDM2G into HEK-293T cells. Furthermore, two stable cell lines expressing luciferase singly or co-expressed luciferase and P21-3'UTR were established by transducing HEK-293 cells with recombinant lentivirus; the former was used as control in the following experiments. Finally luciferase activity of the latter stable cells was measured by using the luciferase reporter assay system. The results showed that high-titre recombinant lentivirus was produced and two stable cell lines were constructed, also to some certain, the luciferase activity was in direct proportion to the number of cells. In conclusion, the high-throughput luciferase reporter system is established successfully; using this system, the 28 miRNA that directly target P21 Cip1/Waf1 are screened experimentally.
Genes, Reporter ; Genetic Vectors ; HEK293 Cells ; Humans ; Lentivirus ; genetics ; Luciferases ; genetics ; MicroRNAs ; genetics ; Plasmids ; Transduction, Genetic

This study was aimed to quantitatively detect the expression level of lysophosphatide acid acyltransferase β (lpaat β) mRNA in leukemia patients so as to provide theoretical basis for the target therapy of lpaat β in leukemia. Real-time fluorescently quantitative PCR was used to detect the relative expression level of lpaat β mRNA to analyze its expression change in various type of leukemia. The results showed that the lpaat β mRNA expression level in acute leukemia (AL) patients was significantly higher than that in normal controls (p < 0.05); lpaat β mRNA expression level in acute myeloid leukemia (AML) patients was significantly higher than that in normal controls (p < 0.05) and was positively correlated with white blood cell count (≥ 20.0 × 10(9)/L) (p < 0.05) and CD34 expression level of leukemia, but was not related with extramedullary infiltration. Except for acute promyelocytic leukemia (APL), the lpaat β mRNA expression level was negatively correlated with chemotherapy sensitivity in chronic myeloid leukemia (AML) patients. lpaat β mRNA expression level in chronic myeloid leukemia (CML) patients was significantly higher than that in normal controls (p < 0.05). lpaat β mRNA expression level in acute lymphoid leukemia (ALL) patients was not higher than that in normal controls (p > 0.05). It is concluded that the lpaat β gene overexpression exists in both AML and CML patients. lpaat β produced by AML cells probably plays an important role in abnormal proliferation and drug-resistance of AML cells.
Acyltransferases ; genetics ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Child ; Female ; Gene Expression ; Humans ; Leukemia ; genetics ; pathology ; Male ; Middle Aged ; RNA, Messenger ; genetics ; Young Adult

This study was aimed to quantitatively detect the levels of april mRNA expression in leukemia patients so as to provide theoretical basis for the target therapy directing at april in leukemia. Real time fluorescent quantitative PCR was used to detect the relative expression level of april mRNA in newly diagnosed leukemia patients and to analyze the changes of its expression level in various type of leukemia. The results showed that the april mRNA expression level in acute leukemia (AL) patients was significantly higher than that in normal controls, there was statistical difference between them (p < 0.05); april mRNA expression level in acute myeloid leukemia (AML) patients was significantly higher than that in normal controls (p < 0.05) and positively correlated with white blood cell count ≥ 20.0 × 10(9)/L (p < 0.05), but not related with extramedullary infiltration and the expression of CD34. Except for acute promyelocytic leukemia (APL), april mRNA expression level was negatively correlated with sensitivity of patients to chemotherapy. april mRNA expression levels in acute lymphoid leukemia (ALL) and chronic myeloid leukemia (CML) patients were not higher than that in normal controls, there was no statistical difference between them (p > 0.05). It is concluded that april gene overexpression exits in AML patients. APRIL protein produced by AML cells probably plays an important role in abnormal proliferation and drug-resistance of AML cells.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Female ; Humans ; Leukemia ; genetics ; therapy ; Male ; Middle Aged ; RNA, Messenger ; genetics ; Tumor Necrosis Factor Ligand Superfamily Member 13 ; genetics ; Young Adult

Objective To evaluate the effect of early mechanical ventilation on the clinical outcome of the patients with severe brain injury. Methods In 251 patients sustaining severe brain injuries with Glasgow Coma Scores(GCS)of 5 to 8,early mechanical ventilation was administered in 128 patients,with the other 123 patients serving as the control group.The oxygen saturation(SaO2),oxygen pressure(PaO2),carbon dioxide pressure(PaCO2),GCS score,heart rate(HR),andblood pressure (BP)were measured before and after the ventilation in the two groups,and the prognosis of the patients were evaluated.Results Before the treatment,the two groups showed comparable SaO2,PaO2,PaCO2,BP,HR and GCS scores(P>0.05).Compared with the control group,early mechanical ventilation significantly improved the blood gas parameters 24 h and 7 and 14 days after the treatment. SaO2 and PaO2 showed significant increases(P<0.05)and PaCO2 decreased significantly(P<0.05)after early ventilation,which resulted in no significant changes in the HR and BP(P>0.05). Conclusion Early mechanical ventilation can significantly improve the blood gas parameters and the clinical outcome of the patients with severe brain injury.