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1.
Eur J Haematol ; 103(3): 215-224, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31206203

RESUMO

AIM: This study aimed to investigate the possible functions of interaction between JARID1B and miR-137 in ALL. METHODS: The levels of H3K4me3 and H3K4me2 and the expression of JARID1B and miR-137 were analyzed in six ALL cell lines and 30 ALL patients. The effects of miR-137 and JARID1B on cell proliferation and apoptosis were investigated by silencing or promoting the respective genes. The interaction between miR-137 and JARID1B was confirmed by double-luciferase report assay. RESULTS: The histone H3K4 expressions and miR-137 expression were lower in 30 ALL patients and in six ALL cell lines, while the expression of JARID1B was elevated. A negative correlation was observed between JARID1B and miR-137. Over-expression of miR-137 led to decreasing cell proliferation and increasing apoptosis in MOLT-4 and BALL-1 cells. MiR-137 inhibitor up-regulated JARID1B in these two cell lines, while promoted proliferation in BALL-1 cells only. Dual-luciferase report assay suggested that JARID1B was a direct target of miR-137 in ALL cell lines. CONCLUSIONS: The expression of miR-137 was declined in ALL, and JARID1B was directly repressed by miR-137. Aberrant JARID1B expression could result in abnormal histone methylation, which might be one cause of ALL.


Assuntos
Regulação Leucêmica da Expressão Gênica , Histona Desmetilases com o Domínio Jumonji/genética , MicroRNAs/genética , Proteínas Nucleares/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Interferência de RNA , Proteínas Repressoras/genética , Regiões 3' não Traduzidas , Adolescente , Adulto , Idoso , Apoptose/genética , Linhagem Celular Tumoral , Proliferação de Células , Feminino , Genes Reporter , Histonas/metabolismo , Humanos , Histona Desmetilases com o Domínio Jumonji/metabolismo , Masculino , Pessoa de Meia-Idade , Proteínas Nucleares/metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras/metabolismo , Proteínas Repressoras/metabolismo , Adulto Jovem
2.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 32(2): 470-475, 2024 Apr.
Artigo em Zh | MEDLINE | ID: mdl-38660854

RESUMO

OBJECTIVE: To investigate the influence of novel CRM1 inhibitor KPT-330 on the autophagy of mantle cell lymphoma (MCL) cells, and effect of KPT-330 on the prolifiration of MCL cells in the presence or absence of autophagy inhibitor. METHODS: CCK-8 assay was used to detect the effect of KPT-330 on MCL cell lines Z-138, Jeko-1, Granta-519, Rec-1. The effect of KPT-330 on autophagy features were determined by detecting acidic vesicular organelles (AVO) by MDC staining under fluorescence microscope and detecting protein expression of LC3B-II assessed by Western blot. Further combined application of lysosomal inhibitor Chloroquine (CQ) was used to observe its effect on the increase of LC3B-Ⅱ caused by KPT-330. CalcuSyn 2.0 software was used to detected the Combination index (CI) of KPT-330 combined with CQ. RESULTS: The proliferation of MCL cell lines (Z-138, Jeko-1, Grant-519, Rec-1) could be inhibited by KPT-330 in a dose-dependent manner (r =0.930, 0.946, 0.691, 0.968 respectively). The number of acidic vesicular organelles (AVO) and the expression of LC3B-II were increased in KPT-330 treated Jeko-1 and Granta-519 cells in a dose-dependent manner (r Jeko-1=0.993, r Granta-519=0.971). LC3B-II protein amounts still increased upon KPT-330 treatment with the existence of lysosomal inhibitor CQ in Jeko-1 and Granta-519 cells, which was higher than CQ or KPT-330 single drug group. The combination of KPT-330 and CQ produced the synergistic effects on cells proliferation inhibition with CalcuSyn 2.0 analysis. CONCLUSION: KPT-330 can inhibit MCL cell proliferation and induce autophagy. KPT-330 combined with autophagy inhibitor CQ could produce synergistic anti MCL effects, providing experimental basis for clinical combination therapy.


Assuntos
Autofagia , Proliferação de Células , Linfoma de Célula do Manto , Linfoma de Célula do Manto/tratamento farmacológico , Humanos , Autofagia/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Cloroquina/farmacologia
3.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 31(3): 722-729, 2023 Jun.
Artigo em Zh | MEDLINE | ID: mdl-37356932

RESUMO

OBJECTIVE: To analyze the clinical characteristics of the patients with B-cell chronic lymphoproliferative disease(B-CLPD) in the new drug era and the effect of new drug treatment on efficacy and survival. METHODS: The clinical and laboratory data of 200 cases B-CLPD patients diagnosed between April 2015 and August 2021 were analyzed retrospectively. The clinical efficacy and survival of the patients under different treatments including Bruton tyrosine kinase(BTK) inhibitors, rituximab, and chemotherapy alone were analyzed. The prognostic factors affecting the survival of patients were analyzed by univarite analysis and multivariate analysis. RESULTS: There were 119 male(59.5%) and 81 female(40.5%) in 200 cases B-CLPD patients, the sex ratio(male/female) was 1.5∶1 with median age of 61(30- 91) years old. The distribution of subtypes were as fallows: 51 cases (25.5%) of chronic lymphocytic leukemia/small lymphocytic lymphoma(CLL/SLL), 64(32.0%) cases of follicular lymphoma(FL), 40(20.0%) cases mantle cell lymphoma(MCL), 30(15.0%) cases of marginal zone lymphoma(MZL), 10(5%) cases of lymphoplasmacytic lymphoma/waldenstrom macroglobulinemia(LPL/WM), 5(2.5%) cases of B cell chronic lymphoproliferative disorders unclassified(B-CLPD-U) . The main clinical manifestation of 102 patients was lymph node enlargement, 32 cases were complicated with B symptoms. Among CLL/SLL patients, there were 12(23.5%) cases in Binet A and 39(76.5%) cases in Binet B/C. There were 29 patients(20.9%) in Ann Arbor or Lugano stage I-II and 110 cases(79.1%) in stage III-IV of other subtypes. The complete remission(CR) rate was 43.1%(25/58), 40.2%(39/97), 7.1%(1/14), and overaIl response rate(ORR) was 87.9%(51/58), 62.9%(61/97), 28.6%(4/14) in the groups of BTK inhibitors, rituximab-based therapy, and chemotherapy alone. The 3-year OS rate and PFS rate in all patients was 79.2% and 72.4% respectively. The 3-year OS rate of patient with MZL, CLL/SLL, FL,WM was 94.7%, 87.7%, 86.8% and 83.3% respectively, while the 3-year OS rate of MCL was only 40.6%, which was significantly lower than other subtypes. The median OS of patients treated with BTK inhibitors and rituximab-based therapy was 20.5 and 18.5 months respectively, and the 3-year OS rate was 97.4% and 90.7%. However, the median PFS of patients receiving chemotherapy alone was 4 months, and the 1-year OS rate was 52.7%, which was statistically significant compared with the other two groups(P<0.05). Univarite analysis showed that anemia, elevated lactate dehydrogenase, elevated ß2-microglobulin, and splenomegaly were the poor prognostic factors for OS(P<0.05), elevated lactate dehydrogenase was also poor prognostic factors for PFS(P<0.05). Multifactor analysis showed that anemia and elevated lactate dehydrogenase were the independent poor prognostic factors for survival(P<0.05). CONCLUSION: The clinical features of B-CLPD was various, anemia and elevated lactate dehydrogenase are the prognostic factors for poor survival. BTK inhibitors and new immunotherapy can improve the survival and prognosis of patients in the new drug era.


Assuntos
Leucemia Linfocítica Crônica de Células B , Linfoma de Zona Marginal Tipo Células B , Linfoma de Célula do Manto , Humanos , Adulto , Feminino , Masculino , Pessoa de Meia-Idade , Idoso , Idoso de 80 Anos ou mais , Rituximab/uso terapêutico , Leucemia Linfocítica Crônica de Células B/tratamento farmacológico , Estudos Retrospectivos , Prognóstico , Lactato Desidrogenases
4.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 28(5): 1570-1577, 2020 Oct.
Artigo em Zh | MEDLINE | ID: mdl-33067956

RESUMO

AbstractObjective:To investigate the expression of miR-215 and KDM1B in DLBCL patients, and to analysis its clinical significance. METHODS: Fifty patients with DLBCL treated in our hospital were selected as DLBCL group, and 30 cases of reactive proliferative lymphadenitis(RPL) were selected as controls. RQ-PCR was used to detect the expression level of miR-215, and immunohistochemistry was used to detect the expression of KDM1B protein. The expression of miR-215 and KDM1B in patients with different clinical characteristics and the survival rate of patients with different expression of miR-215 and KDM1B was compared. miR-215 mimics was transfected into SU-DHL-4 cells. Cell proliferation was detected by CCK-8. Cell apoptosis was measured by flow cytometry. The expression of KDM1B protein was detected by Western blot. RESULTS: The expression of miR-215 in DLBCL patients was significantly lower than that in control group, and the positive expression of KDM1B protein was higher, the difference was statistically significant(P<0.01). Spearman rank correlation analysis showed that the expression of miR-215 negatively correlated with KDM1B (r=-0.751,P<0.05). There was significant correlation of miR-215, KDM1B expression with symptoms of B,Serum level of LDH, International Prognostic Index(IPI), Ann Arbor stage,Tumor size, respectively in patients(P<0.05). Kaplan-Meier showed that 5-year overall survival rate of patients with high miR-215 expression was significantly longer than that with low miR-215 expression (P=0.013). The 5-year survival rate of the patients with high positive KDM1B expression was significantly lower than that with low positive expression(P=0.024). KDM1B protein was suppressed by the transfection of miR-215 mimics for 72 h, the cell proliferation rate in miR-215 mimics group was significantly lower than that in control group and NC mimics group (P<0.05), but cell apoptotic rate of miR-215 mimics were significantly higher(P<0.05). The expression of KDM1B protein was significantly lower than that in control and NC group(P<0.05). CONCLUSION: There are low expression of miR-215 and high expression of KDM1B protein in patients with DLBCL, suggesting that they may be the diagnostic and prognostic indicators of DLBCL. miR-215 can directly target KDM1B to inhibit cell growth and induce apoptosis.


Assuntos
Linfoma Difuso de Grandes Células B , MicroRNAs , Apoptose , Proliferação de Células , Humanos , Oxirredutases N-Desmetilantes , Prognóstico
5.
Yao Xue Xue Bao ; 44(4): 350-4, 2009 Apr.
Artigo em Zh | MEDLINE | ID: mdl-19545050

RESUMO

This study is to investigate the effect of phenylhexyl isothiocyanate (PHI), which has been proved to be a novel histone deacetylase inhibitor (HDACi) recently, on gene p15 de novo expression in acute leukemia cell line Molt-4, and to further study its potential mechanism. Modified methylation specific PCR (MSP) was used to screen p15-M and p15-U mRNA. DNA methyltransferasel (DNMT1), 3A (DNMT3A), 3B (DNMT3B) and p15 mRNA were measured by RT-PCR. P15 protein was detected by Western blotting. Hypermethylation of gene p15 was reversed and activation transcription of gene p15 in Molt-4 was de novo after 5 days exposure to PHI in a concentration dependent manner. DNMT1 and DNMT3B were inhibited by exposure to PHI for 5 days (P < 0.05). Alteration of DNMT3A was not significant. It is showed that PHI could reverse hypermethylation of gene p15 and transcriptional activation of gene p15 is de novo by PHI. It may result from down-regulating DNA methyltransferases, DNMT1 and DNMT3B, or up-regulating the histone acetylation that allows chromatin unfolding and the accessibility of regulators for transcriptional activation in the p15 promoter.


Assuntos
Inibidor de Quinase Dependente de Ciclina p15/genética , DNA (Citosina-5-)-Metiltransferases/metabolismo , Metilação de DNA , Isotiocianatos/farmacologia , Leucemia-Linfoma Linfoblástico de Células T Precursoras , Linhagem Celular Tumoral , Inibidor de Quinase Dependente de Ciclina p15/metabolismo , DNA (Citosina-5-)-Metiltransferases/genética , DNA Metiltransferase 3A , Inibidores de Histona Desacetilases/farmacologia , Humanos , Leucemia-Linfoma Linfoblástico de Células T Precursoras/genética , Leucemia-Linfoma Linfoblástico de Células T Precursoras/metabolismo , Leucemia-Linfoma Linfoblástico de Células T Precursoras/patologia , RNA Mensageiro/metabolismo , Proteínas Repressoras/genética , Proteínas Repressoras/metabolismo , DNA Metiltransferase 3B
6.
Oncol Rep ; 41(1): 377-386, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30365139

RESUMO

MAPK kinase 1 (MEK1) is an upstream protein kinase of extracellular signal regulated kinase (ERK), which activates the ERK/MAPK (mitogen activated protein kinase) pathway. Importantly, bioinformatic analysis has shown that there is a target complementary binding site between miR­101 and MEK1. The present study aimed to ascertain whether or not miR­101 plays a role in regulating MEK1 expression, ERK/MAPK pathway activity, and the proliferation and apoptosis in a diffuse large B cell lymphoma (DLBCL) cell line. DLBCL tumor samples were collected from patients in our hospital, and lymphatic tissues with reactive lymphoid hyperplasia were selected as controls. The patients were divided into high and low expression groups, and then the survival rate of the two groups was compared using Kaplan­Meier method, as well the effect of miR­101 and MEK1 mRNA expression on survival and prognosis was analyzed. The expression of miR­101, MEK1 and p­MEK1 between normal lymphoblastic cell lines (HCC1954 BL and NCI­BL2009) and lymphoma cell lines (SU­DHL­4 and Farage) was compared. Lymphoma SU­DHL­4 and Farage cells were cultured in vitro, and then divided into the following groups: miR­NC group; miR­101 mimic group; siRNA­NC group; and siRNA­MEK1 group. The expression of miR­101, MEK1, p­MEK1, p­ERK1/2 and Bcl­2 was compared. Cell apoptosis was detected by flow cytometry, and cell proliferation was detected by EdU staining. The results showed that targeted regulation existed between miR­101 and MEK1, and the decreased expression of miR­101 was related to the pathogenesis and prognosis of DLBCL. Upregulation of miR­101 inhibited DLBCL cell proliferation and facilitated apoptosis by inhibiting the expression of MEK1.


Assuntos
Apoptose/genética , Proliferação de Células/genética , Linfoma Difuso de Grandes Células B/genética , MAP Quinase Quinase 1/genética , Sistema de Sinalização das MAP Quinases/genética , MicroRNAs/genética , Proteínas Quinases Ativadas por Mitógeno/genética , Adulto , Idoso , Linhagem Celular , Linhagem Celular Tumoral , Feminino , Células HEK293 , Humanos , Linfoma Difuso de Grandes Células B/patologia , Masculino , Pessoa de Meia-Idade , RNA Interferente Pequeno/genética , Transdução de Sinais/genética , Taxa de Sobrevida , Adulto Jovem
7.
Cancer Manag Res ; 11: 2739-2746, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31040714

RESUMO

BACKGROUND: miR-101 is reported to be associated with cell proliferation and apoptosis. However, it is unknown whether miR-101 expression affects cell proliferation and apoptosis in diffuse large B cell lymphoma (DLBCL). The aim of the present study was to investigate the expression of miR-101 and its effect on cell proliferation and apoptosis in DLBCL. METHODS: miR-101 expression was detected in 30 cases of patients with DLBCL and normal lymph node by qRT-PCR. Then, miR-101 expression was up-regulated and down-regulated in Originated Cell Line-Large Lymphoma 8 (OCL-LY8) cell line, respectively. MTT and flow cytometry assay were used to evaluate the effect of miR-101 on cell proliferation and apoptosis, respectively. As KDM1A was confirmed to be as a specific target of miR-101 by TargetScanHuman, the relationship between MiR-101 and KDM1A was further investigated. RESULTS: miR-101 expression in patients with DLBCL was significantly reduced compared those in normal lymph node (P<0.05). miR-101 expression was significantly associated with tumor size, clinical stage and International Prognostic Index (IPI) scores (P<0.05). In OCL-LY8 cell line, miR-101 down-regulation significantly promoted cell proliferation and suppressed cell apoptosis. Meanwhile, miR-101 up-regulation reversed this effect. In addition, miR-101 negatively regulated the expression of KDM1A. KDM1A down-regulation was oberved in normal tissues compared with those in DLBCL tissues, which inhibited cell proliferation and promoted cell apoptosis. CONCLUSION: These data indicate that miR-101 regulates cell proliferation and apoptosis by targeting KDM1A, which provides a potential therapeutic for DLBCL patients.

8.
DNA Cell Biol ; 36(12): 1168-1177, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29058460

RESUMO

Ginsenoside, a natural triterpenoid saponin, exhibits immunomodulatory and anticancer activities. In the present study, we demonstrated that ginsenoside Rg1 induced secretion of cytokines, including interleukin (IL)-6, tumor necrosis factor-alpha (TNF-α), and IL-1ß, and chemokines such as IL-8 and IP-10 in a dose-dependent manner by human peripheral blood mononuclear cell (PBMC)-derived dendritic cells. Rg1 stimulated the expression of the surface molecules CD83, CD80, and human leukocyte antigen - antigen D related (HLA-DR) and decreased the expression of CD14. In in vivo experiments, C57BL/6 mice were divided into four groups, immunized with ovalbumin (OVA), OVA plus Rg1, Rg1, and phosphate-buffered saline (PBS), respectively. Splenocytes from C57BL/6 mice immunized with OVA plus Rg1 produced more antigen-specific splenocyte proliferation activity. The level of IFN-γ and IL-4 in the splenocytes was also upregulated when in vitro stimulated with OVA257-264 or OVA. After in vivo injection of tumor-forming E.G7-OVA cells, the survival rate of mice immunized intraperitoneally in OVA plus Rg1 immunized mice was higher than that in OVA immunized mice or PBS immunized mice. Thus, Rg1 induced a potent vaccine adjuvant effect and elicited antitumor immunity that polarized a Th1 type immune response. Rg1 could have potential as a prophylactic vaccine adjuvant to control lymphomas.


Assuntos
Adjuvantes Imunológicos/administração & dosagem , Vacinas Anticâncer/administração & dosagem , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Ginsenosídeos/administração & dosagem , Animais , Linhagem Celular Tumoral , Citocinas/biossíntese , Humanos , Linfoma/imunologia , Linfoma/patologia , Linfoma/terapia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Ovalbumina/imunologia
9.
Zhonghua Xue Ye Xue Za Zhi ; 37(1): 56-60, 2016 Jan.
Artigo em Zh | MEDLINE | ID: mdl-26876255

RESUMO

OBJECTIVE: To investigate the effect of silencing LSD1 gene by RNA interference on the proliferation, apoptosis on human lymphocytic leukemia Jurkat cell line and its mechanism. METHODS: The hairpin- like oligonucleotide sequences targeting LSD1 gene was transfected into Jurkat cells by lipofectamine(TM) 2000. The LSD1 mRNA and protein were detected by RQ- PCR and Western blot. Cell growth was determined by MTT. Cell apoptosis was analyzed by flow cytometry. The expression of Bcl-2, Bax, procaspase- 3, and histone H3K4me, H3K4me2, H3K4me3, Act- H3, H3K9me were detected by Western blot. RESULTS: LSD1 mRNA was markedly suppressed by the shRNA targeting LSD1. LSD1 shRNA suppressed the proliferation and induced cells apoptosis of Jurkat cells. The cell apoptotic rate was (41.34±3.58)%, (3.45±1.54)%, (1.76±0.52)% in LSD1 shRNA, Neg-shRNA and Blank respectively, the difference among them was statistically significant (P<0.05). LSD1 shRNA down- regulated the expressions of Bcl- 2 and procaspase- 3, and up- regulated the expression of Bax. The methylation of H3K4me1, me2 and acetylation of Act- H3 improved without change of the methylation of H3K4me3. CONCLUSIONS: Deplete of LSD1 gene maybe through modifying the methylation of histone H3K4 to promote the cell apoptosis and inhibit cell growth in Jurkat cell line.


Assuntos
Apoptose , Histona Desmetilases/genética , Interferência de RNA , Acetilação , Caspase 3/metabolismo , Ciclo Celular , Linhagem Celular Tumoral , Proliferação de Células , Regulação para Baixo , Histonas/metabolismo , Humanos , Células Jurkat , Metilação , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , RNA Mensageiro , RNA Interferente Pequeno , Transfecção
10.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 24(3): 722-6, 2016 Jun.
Artigo em Zh | MEDLINE | ID: mdl-27342498

RESUMO

OBJECTIVE: To summarize the clinical features and therapy experience of a case of CD5 positive diffuse large B cell lymphoma (CD5+ DLBCL) with autoimmune hemolytic anemia (AIHA). METHODS: A 49-years old patient was investigated. The routine blood examination, bone marrow smear, Coombs test, serological test, chest CT, abdominal MR and immunochemistry etc were performed for this patient; and therapeutic effects of the chemotherapy regimen consisting of rituximab plus autologous hematopoietic stem cell transplantation (auto-HSCT) were observed. RESULTS: The cervical lymphnode biopsy confirmed CD5+ DLBCL; the severe anemia, reticulocyte increase, Coombs test positive, and erythroid hyperplasia in bone marrow all suggested the occurence of autoimmune hemolytic anemia (AIHA). After plasma exchange, immune suppression using methylprednisolone, blood transfusion, one course of chemotherapy with "R-CHOP-E", the symptoms of AIHA in patient disappeared. After a continuous treatment for 3 courses of "R-CHOP-E", the bone marrow infiltration appeared, which was assessed as "PD", then the treatment was changed to the "R-ESHAP" for 4 courses, the patient was reassessed as "CR". The patient subsequently underwent auto-HSCT, followed up for 6 months, patientis still "CR". CONCLUSION: The status of the CD5+ DLBCL patient with AIHA is severe, and the prognosis is poor. The curative effect of the chemotherapy regimen with rituximab plus auto-HSCT for this patien is well.


Assuntos
Anemia Hemolítica Autoimune/terapia , Transplante de Células-Tronco Hematopoéticas , Linfoma Difuso de Grandes Células B/terapia , Rituximab/uso terapêutico , Anticorpos Monoclonais Murinos/uso terapêutico , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Antígenos CD5/metabolismo , Cisplatino/uso terapêutico , Ciclofosfamida/uso terapêutico , Citarabina/uso terapêutico , Doxorrubicina/uso terapêutico , Etoposídeo/uso terapêutico , Humanos , Metilprednisolona/uso terapêutico , Pessoa de Meia-Idade , Prednisona/uso terapêutico , Biópsia de Linfonodo Sentinela , Vincristina/uso terapêutico
11.
Zhonghua Xue Ye Xue Za Zhi ; 36(1): 49-52, 2015 Jan.
Artigo em Zh | MEDLINE | ID: mdl-25641147

RESUMO

OBJECTIVE: To investigate the effect of silencing mTOR gene by RNA interference on proliferation and apoptosis, and its mechanism on mantle cell lymphoma Jeko-1 cell Line. METHODS: The hairpin-like oligonucleotide sequences targeting mTOR gene was designed and transfected into Jeko-1 cells by lipofectamine TM 2000. The mTOR mRNA and protein were detected by RQ-PCR and Western blot. Cell growth was determined by MTT. Cell apoptosis was analyzed by flow cytometry. The expressions of Bcl-2, Bax, procaspase-3, procaspase-9, P70S6K,and p-P70S6K were detected by Western blot. RESULTS: mTOR mRNA was markedly suppressed by shRNA targeting mTOR. mTOR shRNA suppressed proliferation and induced cells apoptosis of Jeko-1 cells. The cell apoptotic rates were (36.62 ± 3.24)%, (2.58 ± 1.04)%, (1.24 ± 0.30)% respectively, in mTOR shRNA, Neg-shRNA and Blank with statistically significant difference among them (P<0.05). mTOR shRNA down-regulated the expressions of Bcl-2, proCaspase3, proCaspase9 and p-70S6K, up-regulated the expression of Bax. CONCLUSION: Deplete of mTOR gene may be realized through inhibiting the Akt/mTOR signaling pathway to promote the cell apoptosis and inhibit cell growth in Jeko-1 cell line.


Assuntos
Proliferação de Células , Interferência de RNA , Apoptose , Caspase 3 , Caspase 9 , Ciclo Celular , Linhagem Celular Tumoral , Regulação para Baixo , Humanos , Linfoma de Célula do Manto , RNA Mensageiro , RNA Interferente Pequeno , Transdução de Sinais , Serina-Treonina Quinases TOR , Transfecção
12.
Zhonghua Xue Ye Xue Za Zhi ; 34(5): 426-9, 2013 May.
Artigo em Zh | MEDLINE | ID: mdl-23688755

RESUMO

OBJECTIVE: To study the serum protein differential expression in acute leukemia patients and healthy control by differential protein mass spectrometry. METHODS: Serum proteins of 51 acute leukemia (AL) patients and 10 healthy donors were extracted from their peripheral blood. After removing high abundance protein, serum low abundance proteins were separated by two dimensional gel electrophoresis, the differences of serum proteins in AL patients and healthy human were identified. The protein spots with differential expression were cut out and then undergone bleaching, gel digestion and peptide extraction. The peptide mass fingerprint analysis was performed by using MALDI TOF/TOF MS. The protein database MSDB Masort retrieval program was used to evaluate the results. RESULTS: Using Student's t test,19 statistically significant abnormal expression proteins in the serum of AL patients were found compared with the healthy controls (P < 0.05). The expression of α1-trypsin inhibitor (P < 0.01), prealbumin (P < 0.01), trypsin inhibitor (P < 0.01), apolipoprotein E (P < 0.01) and apolipoprotein A-Ⅳ (P < 0.01) decreased, while retinol binding protein (P < 0.05), globin HP2 (P < 0.05), serum lectin (P < 0.05), H factor homologue protein (P < 0.05) and serum amyloid A1 (P < 0.01) increased. Further stratified analysis found that high serum lectin expression in AL patient resulted in poor outcomes. CONCLUSION: There are a variety of serum proteins with differential expression in peripheral blood of AL patients. The differential expression of serum lectin is related to the therapeutic effect. The differential expression of these proteins can be used as a new diagnosis marker or prognostic indicator for acute leukemia.


Assuntos
Proteínas Sanguíneas/metabolismo , Leucemia/sangue , Doença Aguda , Adolescente , Adulto , Idoso , Estudos de Casos e Controles , Feminino , Humanos , Leucemia/diagnóstico , Masculino , Pessoa de Meia-Idade , Mapeamento de Peptídeos , Proteoma/metabolismo , Adulto Jovem
13.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 18(3): 583-7, 2010 Jun.
Artigo em Zh | MEDLINE | ID: mdl-20561406

RESUMO

This study was aimed to investigate the regulatory effect of phenylhexyl isothiocyanate (PHI) on methylation of histone H3K4, H3K9 and demethylation of p15 gene in acute leukemia cell line Molt-4, and to explore the possible mechanism inducing re-expression of silent gene. The methylation status of histone H3K4, H3K9 and the expression of P15 protein in the Molt-4 cells treated with PHI were detected by Western blot; the methylation status of p15 gene in the Molt-4 cells before and after treatment with PHI was determined by methylation specific polymerase chain reaction (MSP); the expression level of p15 gene mRNA in Molt-4 cells treated with PHI was assayed by semiquantitative reverse transcription-PCR. The results indicated that the PHI could increase methylation of histone H3K4 and decrease methylation of histone H3K9 in concentration-and time-dependent manners. After treatment of Molt-4 cells with PHI for 5 days, the methylation of p15 gene was reduced, the significant hypermethylation of p15 gene was reversed, the silenced p15 gene re-expressed; the expressions of p15 mRNA and P15 protein were enhanced in concentration-dependent manner. It is concluded that probably through specifically regulating the methylation level of histone H3K4 and H3K9, the PHI causes the changes of chromosome space structure and results in the demethylation of CPG island in p15 gene, thereby induces the re-expression of p15 gene which was silenced.


Assuntos
Inibidor de Quinase Dependente de Ciclina p15/metabolismo , Metilação de DNA/efeitos dos fármacos , Histonas/genética , Isotiocianatos/farmacologia , Linhagem Celular Tumoral , Ilhas de CpG , Inibidor de Quinase Dependente de Ciclina p15/genética , Regulação Leucêmica da Expressão Gênica/efeitos dos fármacos , Inativação Gênica , Histonas/metabolismo , Humanos
14.
J Hematol Oncol ; 3: 48, 2010 Nov 29.
Artigo em Inglês | MEDLINE | ID: mdl-21114827

RESUMO

BACKGROUND: We have previously demonstrated that phenylhexyl isothiocyanate (PHI), a synthetic isothiocyanate, inhibits histone deacetylases and remodels chromatins to induce growth arrest in HL-60 myeloid leukemia cells in a concentration-dependent manner. METHODS: To investigate the effect of PHI, a novel histone deacetylases inhibitor (HDACi), on demethylation and activation of transcription of p15 in acute lymphoid leukemia cell line Molt-4, and to further decipher the potential mechanism of demethylation, DNA sequencing and modified methylation specific PCR (MSP) were used to screen p15-M and p15-U mRNA after Molt-4 cells were treated with PHI, 5-Aza and TSA. DNA methyltransferase 1 (DNMT1), 3A (DNMT3A), 3B (DNMT3B) and p15 mRNA were measured by RT-PCR. P15 protein, acetylated histone H3 and histone H4 were detected by Western Blot. RESULTS: The gene p15 in Molt-4 cells was hypermethylated and inactive. Hypermethylation of gene p15 was attenuated and p15 gene was activated de novo after 5 days exposure to PHI in a concentration-dependent manner. DNMT1 and DNMT3B were inhibited by PHI (P < 0.05). Alteration of DNMT3A was not significant at those concentrations. Acetylated histone H3 and histone H4 were accumulated markedly after exposure to PHI. CONCLUSION: PHI could induce both DNA demethylation and acetylated H3 and H4 accumulation in Molt-4 cells. Hypermethylation of gene p15 was reversed and p15 transcription could be reactivated de novo by PHI.


Assuntos
Cromatina/genética , Inibidor de Quinase Dependente de Ciclina p15/genética , Metilação de DNA/efeitos dos fármacos , Isotiocianatos/farmacologia , Leucemia de Células T/genética , Acetilação/efeitos dos fármacos , Linhagem Celular Tumoral , Cromatina/efeitos dos fármacos , Cromatina/metabolismo , Inibidor de Quinase Dependente de Ciclina p15/metabolismo , DNA/genética , DNA/metabolismo , Humanos , Leucemia de Células T/metabolismo , Linfócitos T/patologia
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