OBJECTIVETo explore the expression of Bcl-2 mRNA and its effect on prognosis of patients with primary gastrointestinal diffuse large B-cell lymphoma (PGI-DLBCL).

METHODSReal time quantitative PCR was used to determine the expression of Bcl-2 mRNA in 40 PGI-DLBCL patients and 17 healthy controls. The association of Bcl-2 expression with clinicopathological features and prognosis of the patients was analyzed.

RESULTSThe expression level of Bcl-2 mRNA in PGI-DLBCL patients was 1.03 ± 0.93, significantly higher than that of the controls (0.41 ± 0.21) (P < 0.05). The expression of Bcl-2 mRNA in stage IIE-IV patients (1.28 ± 1.01) was significantly higher than that in the stage I-II2 patients (0.62 ± 0.61) (P < 0.05). The expression of Bcl-2 mRNA in patients with international prognostic index (IPI) score >2 (1.95 ± 1.27) was significantly higher than those with IPI score ≤ 2 (0.86 ± 0.75)(P < 0.05). The expression of Bcl-2 mRNA in patients with complete remission (CR) (0.71 ± 0.58) was significantly lower vs. 2.42 ± 0.91 in patients with no CR (P < 0.05). Univariate analysis indicated that β2-MG, IPI score>2, the Lugano staging, and Bcl-2 mRNA expression were associated with overall survival (OS) and progression-free survival (PFS) (P < 0.05). Multivariate analysis indicated that IPI score>2 was independently associated with OS (P < 0.05), and both IPI score >2 and Bcl-2 mRNA expression were independently associated with PFS (P < 0.05).

CONCLUSIONSThe expression of Bcl-2 mRNA in the tumor tissue of PGI-DLBCL patients is significantly higher than that in controls. PGI-DLBCL patients with higher expression of Bcl-2 have a poor chemotherapy response and inferior prognosis. IPI score >2 and higher expression of Bcl-2 mRNA are independent poor prognostic factors for PFS in PGI-DLBCL patients.

Disease-Free Survival ; Genes, bcl-2 ; Humans ; Lymphoma, B-Cell ; diagnosis ; genetics ; metabolism ; Lymphoma, Large B-Cell, Diffuse ; diagnosis ; genetics ; metabolism ; Prognosis ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism

OBJECTIVETo study the expression of miR-16 and bcl-2 in T lymphoblastic lymphoma/leukemia (T-LBL/ALL) and its relationship to prognosis.

METHODS70 cases of T-LBL/ALL with follow-up data were studied by using immunohistochemical EnVision method for CD1a, CD3, cCD3, CD7, CD10, CD20, CD23, CD34, CD43, CD45RO, CD99, TDT, MPO, bcl-2 and Ki-67. The expression levels of miR-16 were examined by TaqMan real-time polymerase chain reaction (RT-PCR). Thirty cases of reactive lymph node were selected as control.

RESULTSAmong the 70 cases of T-LBL/ALL, the percentages of tumor cells expression of TDT, CD99, CD3, CD7, CD10, CD34, CD1a, cCD3, bcl-2, CD45RO and CD43 were 94.3% (66/70), 94.3% (66/70), 68.6% (48/70), 92.9% (65/70), 32.9% (23/70), 24.3% (17/70), 40.0% (28/70), 51.4% (36/70), 34.3% (24/70), 37.1% (26/70), and 48.6% (34/70). Separately, while tumor cells expression of MPO, CD20 and CD23 was all negative. A figure of Ki-67 expression > 80% was found in 24 cases and ≤ 80% in 46 cases. The expression of miR-16 was up-regulated in T-LBL/ALL, and it was 5.07 times of the reactive lymph node(P = 0.001). The high expression group of miR-16 was significantly correlated with longer over survival (P = 0.041). The prognosis of negative bcl-2 group was better than bcl-2 positive one(P = 0.904). The relationship of miR-16 and bcl-2 was significant(P = 0.042,χ(2) = 4.147). Survival multivariate COX proportional hazard regression analysis revealed that the low expression of miR-16 might be a independent poor prognosis factor (P = 0.049).

CONCLUSIONSWhile the high expression group of miR-16 has longer OS than that in low expression group. The prognosis of bcl-2 negative was better than bcl-2 positive. miR-16 may be a independent prognosis factor. The relationship of miR-16 and bcl-2 might suggested that gene regulation may be influenced by them.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Female ; Genes, bcl-2 ; Humans ; Immunohistochemistry ; Infant ; Male ; MicroRNAs ; metabolism ; Middle Aged ; Neoplasm Staging ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; metabolism ; Prognosis ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Survival Rate ; Young Adult

Animals ; Antibodies, Monoclonal, Murine-Derived ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cyclophosphamide ; therapeutic use ; Doxorubicin ; therapeutic use ; Gene Rearrangement ; Genes, myc ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; genetics ; Prednisone ; therapeutic use ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; Proto-Oncogene Proteins c-bcl-6 ; genetics ; metabolism ; Proto-Oncogene Proteins c-myc ; genetics ; metabolism ; Translocation, Genetic ; Vincristine ; therapeutic use

OBJECTIVETo perform immunophenotyping and molecular genetic analysis for diffuse large B-cell lymphoma (DLBCL), and to explore their correlation and implication for prognosis.

METHODSImmunohistochemical streptavidin peroxidase (SP) method was used to determine the expression of CD10, BCL6 and MUM1 in 59 cases of DLBCL. A Hans algorithm was used to classify DLBCL into germinal center B-cell (GCB) and non-GCB subtypes. Interphase fluorescence in situ hybridization (FISH) assay was performed on paraffin-embedded lymphoma tissue sections to detect translocations and amplifications of BCL6, BCL2 and MYC genes with dual-color break-apart BCL6 probe, dual-color dual-fusion IgH/ BCL2 probe and dual-color break-apart MYC probe, respectively.

RESULTSIn the 59 cases of DLBCL, 28.8% (17/59) belonged to GCB subtype, and 71.2% (42/59) belonged to non-GCB subtype. The incidences of BCL6, BCL2 and MYC gene translocations were 24.1% (14/58), 1.7% (1/59) and 5.3% (3/57), respectively. The incidences of BCL6, BCL2 and MYC gene amplifications were 17.2% (10/58), 22.0% (13/59) and 21.1% (12/57), respectively. BCL6 amplification was not correlated with BCL6 translocation (P=0.424), but was correlated with amplifications of BCL2 and MYC (C=0.405 and 0.403, respectively, P <0.01). The incidence of BCL6 translocation in GCB type was higher than that in non-GCB type, and amplifications of BCL6, BCL2 or MYC were more frequently encountered in non-GCB type, though no statistical significance was detected (P=0.089 and 0.106, respectively). By univariate analysis, immunophenotyping and international prognostic index (IPI) exerted a significant effect on overall survival (OS) (P=0.047 and 0.001, respectively), but to which BCL6 translocation and amplification of the 3 genes were not related (P=0.150 and 0.444, respectively). By multivariate analysis, IPI score was the only independent prognostic factor for OS (RR =3.843, P=0.017).

CONCLUSIONThe GCB subtype of DLBCL is less common in the patient cohort. Common genetic aberrations have included BCL6 translocation and BCL6, BCL2 and MYC amplifications. Amplification of the 3 genes is strongly correlated with each other, and the incidence of BCL2 translocation is low. Immunophenotyping only has minor significance for the prognosis. Genetic aberrations cannot predict the clinical outcome of DLBCL.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; DNA-Binding Proteins ; genetics ; Female ; Genes, bcl-2 ; Genes, myc ; Humans ; Immunophenotyping ; In Situ Hybridization, Fluorescence ; Lymphoma, Large B-Cell, Diffuse ; genetics ; immunology ; Male ; Middle Aged ; Proto-Oncogene Proteins c-bcl-6

OBJECTIVETo identify and investigate clinicopathological features of B cell lymphomas with concurrent myc and bcl-2/IgH or bcl-6 translocations ("double-hit" lymphoma).

METHODSTissue microarray was constructed from formalin-fixed and paraffin-embedded tissue samples of aggressive B cell lymphomas diagnosed between 2009 and 2012, including 129 cases of diffuse large B cell lymphoma (DLBCL), 5 cases of B-cell lymphoma, unclassifiable with features intermediate between diffuse large B-cell lymphoma and Burkitt lymphoma (BCLU), 7 cases of Burkitt lymphoma and 4 cases of high-grade follicular lymphoma with diffuse large B cell lymphoma component. Interphase fluorescence in-situ hybridization (FISH) was performed with a panel of probes including myc, bcl-2/IgH and bcl-6 to document related gene translocation and copy number changes. Medical record review was performed and follow-up data was recorded.

RESULTSAmong 145 cases, 5 cases (3.4%) of B cell lymphomas with concurrent myc and bcl-2/IgH or bcl-6 rearrangements (double-hit lymphomas) were identified, including 2 cases involving myc and bcl-2 translocations (1 DLBCL and 1 BCLU), and 3 cases involving myc and bcl-6 translocations (all DLBCLs). Three cases with concurrent bcl-2/IgH and bcl-6 translocations were found. Single gene translocations or increase of copy numbers were found in 66 cases, representing 51.2% (66/129) of all de novo DLBCLs. Ki-67 index of the 5 "double-hit" lymphomas ranged from 60% to 100%. Clinical follow-up data were available in 4 of the 5 "double-hit" lymphoma patients, three of whom died within 2 years and 1 patient was alive after 36 months of follow-up.

CONCLUSIONS"Double-hit" B-cell lymphomas are rare and can only be identified by molecular detection. They should not be considered synonymous with BCLU morphologically, and may present entities within other morphological spectra. Most of the patients have a poor prognosis. Further in-depth studies of larger case numbers are required to determine the pathologic and genetic variables of the lesion.

Adult ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Burkitt Lymphoma ; drug therapy ; genetics ; Cyclophosphamide ; therapeutic use ; Doxorubicin ; therapeutic use ; Female ; Follow-Up Studies ; Genes, bcl-2 ; Genes, myc ; Humans ; In Situ Hybridization, Fluorescence ; Lymphoma, B-Cell ; drug therapy ; genetics ; Lymphoma, Follicular ; drug therapy ; genetics ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; genetics ; Male ; Middle Aged ; Prednisone ; therapeutic use ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; Proto-Oncogene Proteins c-bcl-6 ; genetics ; Proto-Oncogene Proteins c-myc ; genetics ; Retrospective Studies ; Translocation, Genetic ; Vincristine ; therapeutic use

OBJECTIVETo investigate the effect of lentivirus-mediated Bcl-2 gene transfection in protecting human primary ovarian granulosa cells against phosphoramide mustard (PM)-induced apoptosis.

METHODSGranulosa cells were isolated from the follicle fluid of women undergoing in vitro fertilization and embryo transfer. The lentiviral vectors carrying Bcl-2 gene (pGC-FU-Bcl-2) and enhanced green fluorescence protein (pGC-FU-EGFP) were constructed and packaged into high-titer lentiviruses. The resulting recombinant lentivirus carrying Bcl-2 and EGFP genes or the empty vector were used to infect the primary human ovarian granulosa cells, followed by addition of PM in the cell culture, with untreated granulosa cells as the control. The cell apoptosis was detected by Annexin V and Hochst 33258 staining, and the expression of Bcl-2 protein was assessed using Western blotting.

RESULTSThe control granulosa cells showed an apoptotic rate of (1.93±0.28)%. The cells infected with pGC-FU-Bcl-2 prior to PM exposure had a apoptotic rate of (6.99±10.55)%, significantly higher than that of the control cells, but significantly lower than that of the cells with PM exposure only and those infected with the empty vector before PM exposure (P<0.05). The expression of Bcl-2 was the highest in the cells infected with pGC-FU-Bcl-2 prior to PM exposure (P<0.05).

CONCLUSIONLentivirus-mediated Bcl-2 gene transfection can protect human ovarian granulosa cells against PM-induced apoptosis by upregulating Bcl-2 protein expression.

Adult ; Apoptosis ; drug effects ; Female ; Genes, bcl-2 ; Genetic Vectors ; Granulosa Cells ; drug effects ; Humans ; Lentivirus ; genetics ; Phosphoramide Mustards ; Transfection

Adolescent ; Antigens, CD20 ; metabolism ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Burkitt Lymphoma ; drug therapy ; metabolism ; pathology ; surgery ; Child ; Child, Preschool ; Cyclophosphamide ; therapeutic use ; DNA-Binding Proteins ; metabolism ; Doxorubicin ; therapeutic use ; Female ; Follow-Up Studies ; Genes, myc ; Humans ; Hyaluronan Receptors ; metabolism ; Ki-67 Antigen ; metabolism ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; genetics ; metabolism ; pathology ; surgery ; Male ; Neoplasm Staging ; Neprilysin ; metabolism ; Peritoneal Neoplasms ; drug therapy ; genetics ; metabolism ; pathology ; surgery ; Prednisone ; therapeutic use ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Proto-Oncogene Proteins c-bcl-6 ; Stomach Neoplasms ; drug therapy ; genetics ; metabolism ; pathology ; surgery ; Translocation, Genetic ; Vincristine ; therapeutic use

OBJECTIVETo study the cytogenetic profiles of follicular lymphoma (FL) in Chinese patients.

METHODSConventional karyotype in 57 FL patients from Shanghai area was analyzed. Fluorescence in-situ hybridization (FISH) for t(14;18) and Bcl-6 and IgH gene rearrangement was performed in these cases.

RESULTSThe most frequent breakpoints (frequency > or = 10% ) of the 57 FL cases were at band 14q32 (68.4%), 18q21 (38.6%), 3q27 (21.1%), 1q10 (15.8%) and 1q21 (12.3%). Nineteen (33.3%) of the 57 cases had t(14;18). The breakpoint of 18q21 and t(14;18) were more frequent in FL grade 1-2 and less frequent in FL grade 3 (57.6% vs. 12.5%; 54.5% vs. 4.2%, P < 0.05), whereas the 3q27/Bcl-6 rearrangement was more frequent in FL grade 3 and less frequent in FL grade 1-2 (37.5% vs. 9.1% , P < 0.05). The cohort of FL was more frequent in gains of chromosomes X, 1q, 5, 6p, 7 and 12q and losses of chromosomes 1p, 6p and 14q32. Gain of 18q was more frequent in FL grade 1-2 than in FL grade 3 (P < 0.05). Loss of 14q32 was more frequent in t(14;18) negative FL than in t(14;18) positive FL (P < 0.05).

CONCLUSIONSCompared with the data of Western patients reported in the literature, Chinese FL cases had distinct cytogenetic profiles from Western FL cases that the t(14;18) is less frequent and the gain of 1q is more frequent in Chinese FL cases, which are more significant in high grade FL.

Adult ; Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; genetics ; Chromosome Breakage ; Chromosome Deletion ; Chromosomes, Human, Pair 1 ; Chromosomes, Human, Pair 14 ; Chromosomes, Human, Pair 18 ; Female ; Gene Rearrangement ; Genes, bcl-2 ; Humans ; In Situ Hybridization, Fluorescence ; Lymphoma, Follicular ; genetics ; pathology ; Male ; Middle Aged ; Neoplasm Grading ; Translocation, Genetic ; Young Adult

OBJECTIVETo investigate the effect of genistein on osteoblast proliferation, cellular cycle, apoptosis and differentiation of osteoblasts cultivated under hypoxia conditions.

METHODRat osteoblasts were isolated from calvarias by enzyme digestion and a hypoxic model was established by in a triple-gas incubator. Rat osteoblasts were grouped into the normoxic control group, the hypoxia control group and the hypoxia administration group which was subdivided into Ge-6 group, Ge-5 group and Ge-4 group, to which genistein was administered at doses of 1 x 10(-6), 1 x 10(-5), 1 x 10(-4) mol x L(-1). The cell survival rate, lactic dehydrogenase leakage rate, apoptosis and differentiation of osteoblasts were observed for each group at 3 h after hypoxia, and the gene expression of HIF-1alpha, Bcl-2, Caspase-3 was detected by Real time RT-PCR. Forty-eight hours after hypoxia, osteogenic differentiation markers including alkaline phosphatase activity and nodules were detected.

RESULTCompared with the hypoxia control group, the hypoxia administration group displays a significant increase in the survival rate and a decreased in LDH leakage rate, apoptosis rate and percentage of S + G2 phases. Besides, the mRNA level of HIF-1alpha and Bcl-2 were enhanced, the mRNA level of Caspase-3 was inhibited.

CONCLUSIONGenistein has an effect on protecting osteoblasts from hypoxia.

Alkaline Phosphatase ; metabolism ; Animals ; Apoptosis ; drug effects ; Calcification, Physiologic ; drug effects ; Caspase 3 ; genetics ; Cell Cycle ; drug effects ; Cell Hypoxia ; Cell Survival ; drug effects ; Cells, Cultured ; Genes, bcl-2 ; Genistein ; pharmacology ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; L-Lactate Dehydrogenase ; metabolism ; Osteoblasts ; cytology ; drug effects ; metabolism ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley

Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cyclophosphamide ; therapeutic use ; Doxorubicin ; therapeutic use ; Female ; Follow-Up Studies ; Genes, Immunoglobulin Heavy Chain ; Genes, bcl-2 ; Genes, myc ; Humans ; Lymphoma, B-Cell ; drug therapy ; genetics ; pathology ; Male ; Methotrexate ; therapeutic use ; Prednisone ; therapeutic use ; Translocation, Genetic ; Vincristine ; therapeutic use