cyclin-d1 and Lymphoma

The increasing use of immunophenotypic and molecular analysis in the routine evaluation of patients with lymphocytosis, lymphadenopathy, or other hematologic disorders has led to the identification of unexpected small clonal lymphoid populations. These clones, sometimes with disease-specific markers, such as the t(14;18), are especially challenging for the clinician because of their unknown biologic potential and uncertain clinical behavior. Study of these early lymphoid lesions is providing important clues to the process of lymphomagenesis, and may provide the rationale for preemptive therapy in the future. More and more, the hematologist/oncologist is consulted regarding otherwise healthy individuals with lymphadenopathy and/or lymphocytosis, and pathology reports that confound the referring internist or surgeon. The report does not name a malignant lymphoproliferative disorder, but is not completely "normal". Does the patient have a benign or malignant condition? How should they be evaluated? Is treatment indicated? These patients prove challenging for the consulting hematologist as well as the referring physician. In this review, we will focus on some of these scenarios and attempt to provide guidance for their management.

Topics: B-Lymphocytes; Bone Marrow Cells; Cell Transformation, Neoplastic; Cyclin D1; Disease Progression; Hematology; Humans; Immunophenotyping; Lymph Nodes; Lymphatic Diseases; Lymphocytosis; Lymphoma; Lymphoproliferative Disorders; Risk; Translocation, Genetic; Treatment Outcome

Topics: Cyclin D1; Gene Rearrangement; Germinal Center; Humans; Lymphoma; Lymphoma, Follicular; Lymphoma, Mantle-Cell; Proto-Oncogene Proteins c-bcl-2

The clinical usefulness of tumor markers for malignant lymphoma is thought to be in monitoring the therapeutic effect and as a prognostic factor before treatment. The former include specific biological marker such as soluble interleukin-2 receptor measured by ELISA. The latter are cellular prognostic markers detected by immunohistological and flow cytometric analysis. The cyclin D1 over-expression of mantle cell lymphoma in diffuse large B-cell lymphoma, which should be recommended for myeloablative therapy, is a significantly poor prognostic risk factor.

Topics: Biomarkers, Tumor; Cyclin D1; Drug Monitoring; Enzyme-Linked Immunosorbent Assay; Humans; Interleukin-2; Lymphoma; Lymphoma, B-Cell; Lymphoma, Non-Hodgkin; Prognosis; Receptors, Interleukin-2

Cytogenetic analysis of small lymphocytes disorders is hindered by the low mitotic activity of the malignant cells. The use of fluorescence in situ hybridization (FISH) allows the detection of chromosomal amplifications, deletions, or translocations at a single-cell level in dividing and resting cells. The use of FISH in combination with other molecular techniques has defined the deletion in band 13q14 as the most common abnormality in chronic lymphocytic leukemia, followed by del (11)(q22-23), trisomy 12, del (17)(p13), and del (6)(q21). The del 13q14 is also found in 70% of mantle-cell lymphomas (MCLs) and in non-Hodgkin's lymphoma (NHL), acute lymphoblastic leukemia (ALL), and multiple myeloma (MM) patients. These findings point to the existence of yet unidentified tumor-suppressor gene(s) at the 13q14 locus, the loss/inactivation of which leads to B-cell neoplasia. Del (17(p13) (involving the p53 tumor-suppressor gene) and del (11)(q22-23) (involving the ataxia-telangiectasia gene [ATM]) seem to be independent prognostic factors for poor survival in chronic lymphocytic leukemia (CLL) patients. In MCL, the t(11;14) involving the bcl-1 gene is found, but data from a bcl-1 transgenic animal model suggest that hyperexpression of bcl-1 is not sufficient for lymphomatogenesis. Similar data are observed in bcl-2 transgenic animals, a finding showing that the bcl-2 hyperexpression observed in t(14;18)-positive follicular lymphoma cells is not sufficient to confer a malignant phenotype. The contribution of other chromosomal abnormalities other than bcl-1 and bcl-2 rearrangements in the pathogenesis of MCL and follicular-cell lymphomas has to be determined.

Topics: Animals; Animals, Genetically Modified; Chromosome Aberrations; Chromosome Disorders; Cyclin D1; Genes, Tumor Suppressor; Humans; Leukemia, Lymphocytic, Chronic, B-Cell; Lymphoma; Tumor Suppressor Protein p53; Waldenstrom Macroglobulinemia

Topics: Cell Cycle; Cyclin D1; Cyclins; Humans; Lymphoma; Oncogene Proteins

Topics: Chromosomes, Human, Pair 11; Chromosomes, Human, Pair 14; Cyclin D1; Humans; Lymphoma; Protein-Tyrosine Kinases; Proto-Oncogene Proteins; Proto-Oncogenes; Translocation, Genetic

Bardoxolone methyl, a novel synthetic triterpenoid and antioxidant inflammation modulator, potently induces Nrf2 and inhibits NF-κB and Janus-activated kinase/STAT signaling. This first-in-human phase I clinical trial aimed to determine the dose-limiting toxicities (DLT), maximum tolerated dose (MTD), and appropriate dose for phase II studies; characterize pharmacokinetic and pharmacodynamic parameters; and assess antitumor activity.. Bardoxolone methyl was administered orally once daily for 21 days of a 28-day cycle. An accelerated titration design was employed until a grade 2-related adverse event occurred. A standard 3 + 3 dose escalation was then employed until the MTD was reached. Single dose and steady-state plasma pharmacokinetics of the drug were characterized. Assessment of Nrf2 activation was examined in peripheral blood mononuclear cells (PBMC) by measuring NAD(P)H:quinone oxidoreductase (NQO1) mRNA levels. Immunohistochemical assessment of markers of inflammation, cell cycle, and apoptosis was carried out on tumor biopsies.. The DLTs were grade 3 reversible liver transaminase elevations. The MTD was established as 900 mg/d. A complete tumor response occurred in a mantle cell lymphoma patient, and a partial response was observed in an anaplastic thyroid carcinoma patient. NQO1 mRNA levels increased in PBMCs, and NF-κB and cyclin D1 levels decreased in tumor biopsies. Estimated glomerular filtration rate (eGFR) was also increased.. Bardoxolone methyl was well tolerated with an MTD of 900 mg/d. The increase in eGFR suggests that bardoxolone methyl might be beneficial in chronic kidney disease. Objective tumor responses and pharmacodynamic effects were observed, supporting continued development of other synthetic triterpenoids in cancer.

Topics: Adult; Aged; Aged, 80 and over; Antineoplastic Agents; Carcinoma, Renal Cell; Colorectal Neoplasms; Cyclin D1; Dose-Response Relationship, Drug; Drug Administration Schedule; Female; Humans; Janus Kinases; Kidney Neoplasms; Lymphoma; Male; Maximum Tolerated Dose; Melanoma; Middle Aged; NAD(P)H Dehydrogenase (Quinone); Neoplasms; NF-E2-Related Factor 2; NF-kappa B; Oleanolic Acid; RNA, Messenger; STAT Transcription Factors; Thyroid Neoplasms; Young Adult

Pyrazole or 1,2-Diazole is a five-membered heteroaromatic ring with two nitrogen atoms which is widely used in pharmacological research and organic synthesis. Several natural and synthetic pyrazole derivatives possess anti-cancer potential and some of them have underwent clinical trials. In this aspect, an investigation into the efficiency of the pyrazole nucleus to inhibit the growth and progression of various cancer cell lines/ experimental tumours would help in giving a better clarity to the anti-cancer behaviour of pyrazole containing drugs. This paper investigates the efficiency of pyrazole against Dalton's Lymphoma Ascites (DLA) cell line. Pyrazole inhibited the growth of DLA cells in vitro by committing them towards apoptosis. In vitro results were consistent in DLA induced murine solid tumour in vivo systems. Drug-treatment improved survival, reduced tumour loads, stabilized body weights and improved the haematological and serum biochemical parameters of DLA solid tumour bearing mice, thereby improving their overall survivability. Drug administration contained the aggravation of solid tumour by targeted downregulation of Cyclin-D1 and Ki-67. In addition, the mRNA expression levels of anti-apoptotic genes, BCL-2 and BCL-XL were downregulated in solid tumours, corroborating the in vitro results that pyrazole encourage apoptotic cell death in DLA cells. The new findings establish pyrazole as a potential anti-cancer drug candidate. The results must encourage future investigations into the efficacy of the drug against various cancer types.

Topics: Animals; Antineoplastic Agents; Apoptosis; Ascites; bcl-X Protein; Cell Line, Tumor; Cell Proliferation; Cyclin D1; Gene Expression Regulation, Neoplastic; Ki-67 Antigen; Lymphoma; Male; Mice; Mice, Inbred BALB C; Proto-Oncogene Proteins c-bcl-2; Pyrazoles; Signal Transduction

This study aims to evaluate the association between polymorphisms of methotrexate pathway genes and high-dose methotrexate-related hepatotoxicity in Chinese patients with primary central nervous system lymphoma.. Sixty-five patients in 411 treatment courses were enrolled and their toxicities were evaluated. The association between 30 candidate SNPs from 20 methotrexate pathway genes and high-dose methotrexate-related hepatotoxicity was analysed by PLINK and logistic regression.. TYMS 6 bp DI + II (rs151264360; OR, 0.41; 95% CI, 0.25-0.66; P = 0.00029), MTHFD1 1958 GA + AA (rs2236225; OR, 0.55; 95% CI, 0.33-0.91; P = 0.020) and CCND1 870 GA + GG (rs9344; OR, 0.42; 95% CI, 0.24-0.73; P = 0.0024) had less risk of hepatotoxicity compared with their homozygotes (DD, GG and AA, respectively), while ABCC2 intron 29 GA + GG (rs3740065; OR, 3.14; 95% CI, 1.89-5.20; P = 0.00001) was more prevalent in patients with hepatotoxicity than TT.. TYMS 6 bp DI + II, MTHFD1 1958 GA + AA, CCND1 870 GA + GG genotypes were associated with a lower probability of hepatotoxicity in patients with primary central nervous system lymphoma on high-dose methotrexate therapy, and ABCC2 intron 29 GA + GG was correlated with increased risk of hepatotoxicity.

Topics: Adult; Aged; Aminohydrolases; Asian People; Central Nervous System; Chemical and Drug Induced Liver Injury; Cyclin D1; Female; Formate-Tetrahydrofolate Ligase; Genotype; Humans; Introns; Liver; Lymphoma; Male; Methotrexate; Methylenetetrahydrofolate Dehydrogenase (NADP); Middle Aged; Multidrug Resistance-Associated Protein 2; Multienzyme Complexes; Odds Ratio; Polymorphism, Single Nucleotide; Thymidylate Synthase

The present study, attempts to validate the molecular mechanism(s) of Poly-l-lysine (PLL) induced apoptosis, anti-proliferative and anti-tumorigenic properties in in-vitro HUVECs cells and Dalton's Ascitic Lymphoma (DAL) and in in-vivo DAL cell bearing BALB/c mice model.. The cell proliferation assay and morphological assay was carried out using the MTT assay and Giemsa staining method. The antitumor activity of PLL was evaluated in BALB/c mice at 20 and 40 mg/kg/b.w doses for 21 days for DAL solid tumor model. Several tumor evaluation endpoints, hematological and biochemical parameters were estimated. Additionally, the tumor apoptosis, anti-proliferative and anti-tumor angiogenesis effects were assessed using western blots and immunohistochemistry.. PLL significantly decreased cell proliferation in in-vitro HUVECs and DAL cells without significant effects on normal cell growth. PLL also induced alteration in cellular morphology in DAL cells. Therafter, in the BALB/c mouse model, PLL had noticeable inhibition in DAL-induced tumorigenesis. This inhibition was evident through reduced solid tumor volume and weight versus the control group. However, PLL promoted tumor apoptosis and suppressed cell-proliferation and tumor-angiogenesis. PLL also increased hematological markers significantly compared to 5-flurouracil (5-FU). The amount of TdT in the nuclei of DAL cells in mice treated with PLL was significantly increased while in contrast decreases of anti-apoptotic protein Bcl-2 expression were observed. PLL also significantly upregulated the pro-apoptotic protein Bax and activated caspase-3. Measurable decreases of cyclin-D. The present study offers opportunities and hopes for possible anti-tumortherapies with PLL in the near future and warrants further formulation developments.

Topics: Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Ascites; Carcinogenesis; Cell Line, Tumor; Cell Proliferation; Cell Shape; Cyclin D1; Disease Models, Animal; Down-Regulation; Female; Human Umbilical Vein Endothelial Cells; Humans; Kaplan-Meier Estimate; Ki-67 Antigen; Lymphoma; Mice, Inbred BALB C; Neovascularization, Pathologic; Polylysine; Survival Analysis

Protein arginine methyltransferase 5 (PRMT5) has been implicated as a key modulator of lymphomagenesis. Whether PRMT5 has overt oncogenic function in the context of leukemia/lymphoma and whether it represents a therapeutic target remains to be established. We demonstrate that inactivation of PRMT5 inhibits colony-forming activity by multiple oncogenic drivers, including cyclin D1, c-MYC, NOTCH1, and MLL-AF9. Furthermore, we demonstrate that PRMT5 overexpression specifically cooperates with cyclin D1 to drive lymphomagenesis in a mouse model, revealing inherent neoplastic activity. Molecular analysis of lymphomas revealed that arginine methylation of p53 selectively suppresses expression of crucial proapoptotic and antiproliferative target genes, thereby sustaining tumor cell self-renewal and proliferation and bypassing the need for the acquisition of inactivating p53 mutations. Critically, analysis of human tumor specimens reveals a strong correlation between cyclin D1 overexpression and p53 methylation, supporting the biomedical relevance of this pathway.. We have identified and functionally validated a crucial role for PRMT5 for the inhibition of p53-dependent tumor suppression in response to oncogenic insults. The requisite role for PRMT5 in the context of multiple lymphoma/leukemia oncogenic drivers suggests a molecular rationale for therapeutic development.

Topics: Adaptor Proteins, Signal Transducing; Amino Acid Substitution; Animals; Apoptosis; Arginine; Cell Transformation, Neoplastic; Cluster Analysis; Cyclin D1; Cyclin-Dependent Kinase 4; Enzyme Activation; Gene Expression Profiling; Humans; Leukemia, T-Cell; Lymphoma; Lymphoma, T-Cell; Methylation; Mice; Mutation; Oncogenes; Phosphorylation; Protein-Arginine N-Methyltransferases; Tumor Suppressor Protein p53

Cyclin D1 deregulation is implicated in the genesis of multiple human cancers. Importantly, nuclear cyclin D1 retention during S-phase promotes DNA re-replication and subsequent genomic instability, providing a direct correlation between aberrant cyclin D1/CDK4 activity, transcriptional regulation and double strand DNA break (DSB) induction. Together, these molecular events catalyze the genomic instability necessary for neoplastic transformation. Given that replication-associated DNA damage is central to cyclin D1-driven neoplasia, inactivation of critical checkpoint mediators should augment cyclin D1-dependent tumorigenesis in vivo. To interrogate potential synergy between constitutively nuclear cyclin D1 expression and impaired DSB-induced checkpoint integrity, Ataxia Telangiectasia Mutated (ATM)-deficient mice harboring the Eμ-D1T286A transgene were generated and evaluated for tumor onset. Eμ-D1T286A/ATM-/- mice exhibit dramatically accelerated incidence of both B- and T-cell lymphomas relative to Eμ-D1T286A or ATM-/- control cohorts. Lymphomas exhibit clonal chromosomal alterations distinct from ATM-/- mice, which typically acquire translocations involving the Tcrα/δ locus during V(D)J recombination, and instead harbor alterations at the c-Myc locus. Collectively, these findings reveal an intricate relationship wherein nuclear cyclin D1/CDK4 drives genomic instability in the absence of ATM function and clonal selection of cells harboring alterations within the murine c-Myc locus, ultimately facilitating transformation and tumor formation.

Topics: Animals; Ataxia Telangiectasia Mutated Proteins; Carcinogenesis; Cell Nucleus; Chromosome Aberrations; Cyclin D1; Genomic Instability; Humans; Lymphoma; Mice; Mice, Knockout; Protein Transport; Proto-Oncogene Proteins c-myc; Tumor Burden

Evidence is accumulating that hematologic malignancies develop following acquisition of multiple genetic changes. Despite providing many insights into the way by which given genetic changes contribute to the development of disease, the generation of animal models is often laborious. We show a simplified method that allows the retroviral transduction of genes of interest into mouse B or T cells, thus leading to the rapid generation of models of lymphoid neoplasm in mice. Specifically, germinal center B cells induced in vitro from naive mouse B cells and infected with retroviruses for Myc and Bcl2 rapidly developed a neoplasm of immunoglobulin-expressing mature B cells in transplanted mice. Likewise, T cells induced in vitro from immature hematopoietic cells and infected with retroviruses for Myc, Bcl2, and Ccnd1 rapidly developed CD4(+)CD8(-) and CD4(+)CD8(+) T cell neoplasm in transplanted mice. These findings support the use of our simplified method as a versatile tool for lymphoma research.

Topics: Animals; B-Lymphocytes; Cyclin D1; Disease Models, Animal; Genes, bcl-2; Genes, myc; Genetic Vectors; In Vitro Techniques; Lymphoma; Mice; Retroviridae; T-Lymphocytes; Transduction, Genetic

Phosphatidylinositol-3-kinase (PI3K) signaling is constitutive in most human cancers. Selective inhibition of PI3Kδ (p110δ) by GS-1101 has emerged as a promising therapy in chronic lymphocytic leukemia and indolent lymphomas. In aggressive non-Hodgkin lymphomas such as mantle cell lymphoma (MCL), however, efficacy has been observed, but the extent and duration of tumor control is modest. To determine if tumor killing by GS-1101 is cell cycle-dependent, we show in primary MCL cells by whole-transcriptome sequencing that, despite aberrant expression and recurrent mutations in Cyclin D1, mutations are rare in coding regions of CDK4, RB1 and other genes that control G1-S cell cycle progression or PI3K/AKT signaling. PI3Kδ is the predominant PI3K catalytic subunit expressed, and inhibition by GS-1101 transiently inhibits AKT phosphorylation but not proliferation in MCL cells. Induction of prolonged early G1-arrest (pG1) by selective inhibition of CDK4/CDK6 with PD 0332991 amplifies and sustains PI3Kδ inhibition, which leads to robust apoptosis. Accordingly, inhibition of PI3Kδ induces apoptosis of primary MCL tumor cells once they have ceased to cycle ex vivo, and this killing is enhanced by PD 0332991 inhibition of CDK4/CDK6. PIK3IP1, a negative PI3K regulator, appears to mediate pG1 sensitization to PI3K inhibition; it is markedly reduced in MCL tumor cells compared with normal peripheral B cells, profoundly induced in pG1 and required for pG1 sensitization to GS-1101. Thus, the magnitude and duration of PI3K inhibition and tumor killing by GS-1101 is pG1-dependent, suggesting induction of pG1 by CDK4/CDK6 inhibition as a strategy to sensitize proliferating lymphoma cells to PI3K inhibition.

Topics: Cell Cycle; Cell Line, Tumor; Cyclin D1; Cyclin-Dependent Kinase 4; Cyclin-Dependent Kinase 6; G1 Phase Cell Cycle Checkpoints; Humans; Intracellular Signaling Peptides and Proteins; Lymphoma; Membrane Proteins; Mutation; Phosphatidylinositol 3-Kinases; Piperazines; Purines; Pyridines; Quinazolinones

Epigenetic regulation mediated by lysine- and arginine-specific enzymes plays an essential role in tumorigenesis, and enhanced expression of the type II protein arginine methyltransferase PRMT5 as well as the polycomb repressor complex PRC2 has been associated with increased cell proliferation and survival. Here, we show that PRMT5 is overexpressed in three different types of non-Hodgkin lymphoma cell lines and clinical samples as well as in mouse primary lymphoma cells and that it up-regulates PRC2 expression through inactivation of the retinoblastoma proteins RB1 and RBL2. Although PRMT5 epigenetically controls RBL2 expression, it indirectly promotes RB1 phosphorylation through enhanced cyclin D1 expression. Furthermore, we demonstrate that PRMT5 knockdown in non-Hodgkin lymphoma cell lines and mouse primary lymphoma cells leads to RBL2 derepression and RB1 reactivation, which in turn inhibit PRC2 expression and trigger derepression of its CASP10, DAP1, HOXA5, and HRK pro-apoptotic target genes. We also show that reduced PRMT5 expression leads to cyclin D1 transcriptional repression via loss of TP53K372 methylation, which results in decreased BCL3 expression and enhanced recruitment of NF-κB p52-HDAC1 repressor complexes to the cyclin D1 promoter. These findings indicate that PRMT5 is a master epigenetic regulator that governs expression of its own target genes and those regulated by PRC2 and that its inhibition could offer a promising therapeutic strategy for lymphoma patients.

Topics: Animals; Cell Death; Cell Line, Tumor; Cyclin D1; Epigenesis, Genetic; Gene Knockdown Techniques; Genes, Retinoblastoma; Histone Deacetylase 2; Humans; Lymphoma; Lymphoma, Non-Hodgkin; Mice; Polycomb Repressive Complex 2; Promoter Regions, Genetic; Protein Methyltransferases; Protein-Arginine N-Methyltransferases; Retinoblastoma Protein; Retinoblastoma-Like Protein p130; Signal Transduction; Tumor Cells, Cultured

To evaluate the value of cytomorphologic and immunocytochemical approaches in the diagnosis of hematologic neoplasms in serous effusion.. The cytospin and Thinprep smears of effusion specimens were prepared from 23 cases of lymphoid malignancies with histological confirmation and 30 cases of benign effusions used as control. Morphological assessment of the cellular components was conducted, including the ratio of mesothelium to lymphocyte, karyomorphism of lymphoid cell and the presence of apoptosis and mitosis. Immunocytochemical study was performed in all the cases, with flow cytometry in one case.. Among the 23 tumor cases, 14 represented disease relapse, and in the remaining nine cases, the serous effusion was the primary manifestation. The proportion of mesothelium was low in the tumor group, being less than 10% in 20 cases (87.0%, 20/23). It was more than 10% in most of benign cases (20/30, 66.7%). Lymphoid cells were prominent (> 80% cells) in 69.6% of the tumor cases, and the cellular component in some control cases (63.3%, 19/30) showed fewer lymphocytes. Nipple-like projection of lymphocytic nuclei could be detected in almost all the tumor cases (91.3%, 21/23), but was occasionally found in the control group (26.7%, 8/30). Apoptosis and mitosis were obvious in lymphomatous effusion, but observed in only 6.7% of the control cases. Significant difference of the previously mentioned cytomorphologic features existed between the tumor and control groups (P < 0.01). The results of immunocytochemical staining in cell block were identical to the corresponding immunohistochemistry, and one case of mantle cell lymphoma was confirmed by flow cytometry. The cytologic findings seen in all the 23 studied cases were in agreement with the corresponding histologic diagnosis.. Some cytomorphologic features, including decreased number of mesothelium, increased number of lymphoid cells, nuclear nipple-like projection, and the presence of apoptosis and mitosis, are very useful for diagnosing lymphoid malignancy in serous effusion. Immunocytochemistry is an important approach to the cytodiagnosis and classification of lymphoma.

Topics: Adult; Aged; Aged, 80 and over; Apoptosis; Ascitic Fluid; Cyclin D1; Cytodiagnosis; Female; Humans; Immunohistochemistry; Interferon Regulatory Factors; Lymphocytes; Lymphoma; Lymphoma, Large B-Cell, Diffuse; Male; Middle Aged; Mitosis; Pleural Effusion, Malignant; Young Adult

In association to our undestanding of the pathogenesis and biopsy diagnosis of mantle cell lymphoma using immunohistochemical detection of cyclin D1 expression and/or FISH detection of t(11;14)(q13;q32) all the lymphomas interfering with these factors are discussed in a form of a review. This includes a cyclin D1 negative mantle cell lymphoma, as well as other than MCL lymphomas showing positive intranuclear cyclin D1 positivity due to the changes either at transcriptional or postranscriptional levels. In addition to the MCL, the cyclin D1 positivity might be detected in the cells of hairy cell leukemia, plasmocytic lymphoma and diffuse large B-cell lymphoma. In the first two lymphomas the differential diagnostic problems usually do not arise (with exception of G3 plasmacytoma) and cyclin D1 expression might be of interess to understand better their biology, or to represent a prognostically significant factor. In contrast, cyclin D1 positivity in diffuse large B-cell lymphomas demonstrates the possible role of cyclins in the pathogenesis of this lymphoma and may lead to the problems of the differential diagnosis of aggressive variant of pleomorphic MCL (especially when occuring with CD5 positivity coexpression ). The review includes discussion related to the significance of cyclin D1 positivity and to the approach in the immunohistochemical and FISH analysis of the biopsy material.

Topics: Biomarkers, Tumor; CD5 Antigens; Cyclin D1; Diagnosis, Differential; Humans; Lymphoma; Lymphoma, Mantle-Cell

Cyclin D1 elicits transcriptional effects through inactivation of the retinoblastoma protein and direct association with transcriptional regulators. The current work reveals a molecular relationship between cyclin D1/CDK4 kinase and protein arginine methyltransferase 5 (PRMT5), an enzyme associated with histone methylation and transcriptional repression. Primary tumors of a mouse lymphoma model exhibit increased PRMT5 methyltransferase activity and histone arginine methylation. Analyses demonstrate that MEP50, a PRMT5 coregulatory factor, is a CDK4 substrate, and phosphorylation increases PRMT5/MEP50 activity. Increased PRMT5 activity mediates key events associated with cyclin D1-dependent neoplastic growth, including CUL4 repression, CDT1 overexpression, and DNA rereplication. Importantly, human cancers harboring mutations in Fbx4, the cyclin D1 E3 ligase, exhibit nuclear cyclin D1 accumulation and increased PRMT5 activity.

Topics: Adaptor Proteins, Signal Transducing; Animals; Cell Cycle Proteins; Cell Line, Tumor; Cell Nucleus; Cell Proliferation; Cell Survival; Cell Transformation, Neoplastic; Cullin Proteins; Cyclin D1; Cyclin-Dependent Kinase 4; DNA Methylation; DNA Replication; Enzyme Activation; F-Box Proteins; Gene Expression Regulation, Neoplastic; Histones; Humans; Lymphoma; Mice; Neoplasms; Phosphorylation; Promoter Regions, Genetic; Protein Binding; Protein Methyltransferases; Protein Stability

The breaking of peripheral T-cell tolerance toward self-antigens expressed by tumor cells and the subsequent establishment of an effective tumor protective immune response remains a major challenge for cancer immunotherapy. We report that both protective and therapeutic anti-tumor immune responses can be achieved in a mouse leukemia/lymphoma tumor model through the strong adjuvant effects provided by allogeneic CD3/CD28 cross-linked Th1 memory cells. The adjuvant effect of these cells is mediated by their ability to produce a variety of 'danger signals' which serve to deviate native non-protective Th2 anti-leukemia immune responses to effective Th1 immune responses.

Topics: Animals; Antigens, Neoplasm; Cancer Vaccines; CD28 Antigens; CD3 Complex; Cell Differentiation; Cyclin D1; Female; Immunologic Memory; Immunotherapy, Active; Leukemia; Lymphoma; Male; Mice; Th1 Cells; Transplantation, Homologous

Abnormal thymocyte development with thymic lymphomagenesis inevitably occurs in Atm-/- mice, indicating that ATM plays a pivotal role in regulating postnatal thymocyte development and preventing thymic lymphomagenesis. The mechanism for ATM controls these processes is unclear. We have shown previously that c-Myc, an oncoprotein regulated by the mammalian target of rapamycin (mTOR), is overexpressed in Atm-/- thymocytes. Here, we show that inhibition of mTOR signaling with its specific inhibitor, rapamycin, suppresses normal thymocyte DNA synthesis by downregulating 4EBP1, but not S6K, and that 4EBP1 phosphorylation and cyclin D1 expression are coordinately increased in Atm-/- thymocytes. Administration of rapamycin to Atm-/- mice attenuates elevated phospho-4EBP1, c-Myc and cyclin D1 in their thymocytes, and delays thymic lymphoma development. These results indicate that mTOR downstream effector 4EBP1 is essential for normal thymocyte proliferation, but deregulation of 4EBP1 in Atm deficiency is a major factor driving thymic lymphomagenesis in the animals.

Topics: Adaptor Proteins, Signal Transducing; Animals; Antibiotics, Antineoplastic; Ataxia Telangiectasia Mutated Proteins; Carrier Proteins; Cell Cycle Proteins; Cyclin D1; DNA Replication; DNA-Binding Proteins; Eukaryotic Initiation Factors; Lymphoma; Mice; Mice, Knockout; Phosphoproteins; Phosphotransferases (Alcohol Group Acceptor); Protein Serine-Threonine Kinases; Proto-Oncogene Proteins c-myc; Signal Transduction; Sirolimus; Thymus Neoplasms; TOR Serine-Threonine Kinases; Tumor Suppressor Proteins

The process of translation occurs at a nexus point downstream of a number of signal pathways and developmental processes. Modeling activation of the PTEN/AKT/mTOR pathway in the Emu-Myc mouse is a valuable tool to study tumor genotype/chemosensitivity relationships in vivo. In this model, blocking translation initiation with silvestrol, an inhibitor of the ribosome recruitment step has been showed to modulate the sensitivity of the tumors to the effect of standard chemotherapy. However, inhibitors of translation elongation have been tested as potential anti-cancer therapeutic agents in vitro, but have not been extensively tested in genetically well-defined mouse tumor models or for potential synergy with standard of care agents.. Here, we chose four structurally different chemical inhibitors of translation elongation: homoharringtonine, bruceantin, didemnin B and cycloheximide, and tested their ability to alter the chemoresistance of Emu-myc lymphomas harbouring lesions in Pten, Tsc2, Bcl-2, or eIF4E. We show that in some genetic settings, translation elongation inhibitors are able to synergize with doxorubicin by reinstating an apoptotic program in tumor cells. We attribute this effect to a reduction in levels of pro-oncogenic or pro-survival proteins having short half-lives, like Mcl-1, cyclin D1 or c-Myc. Using lymphomas cells grown ex vivo we reproduced the synergy observed in mice between chemotherapy and elongation inhibition and show that this is reversed by blocking protein degradation with a proteasome inhibitor.. Our results indicate that depleting short-lived pro-survival factors by inhibiting their synthesis could achieve a therapeutic response in tumors harboring PTEN/AKT/mTOR pathway mutations.

Topics: Animals; Base Sequence; Cell Line, Tumor; Cyclin D1; Cycloheximide; Depsipeptides; DNA Primers; Drug Resistance, Neoplasm; Eukaryotic Initiation Factor-4E; Female; Genes, bcl-2; Genes, myc; Harringtonines; Homoharringtonine; Lymphoma; Mice; Mice, Inbred C57BL; Mutation; Myeloid Cell Leukemia Sequence 1 Protein; Peptide Chain Elongation, Translational; Proto-Oncogene Proteins c-bcl-2; Proto-Oncogene Proteins c-myc; PTEN Phosphohydrolase; Quassins; Tuberous Sclerosis Complex 2 Protein; Tumor Suppressor Proteins

We examined the effect of apogossypolone (ApoG2), a new derivative from gossypol on cell cycle regulation in U937 human leukemic monocyte lymphoma cells in vitro. ApoG2 decreased the viability of U937 cells by inducing G1 arrest followed by apoptosis in a dose-dependent manner. The G0/G1 phase of the cell cycle is regulated by cyclin-dependent kinases (Cdk), cyclins and cyclin-dependent kinase inhibitors (Cdki). We show by western blot analysis, that the ApoG2-induced G1 arrest was mediated through the increased expression of Cdki proteins (p21cip1/waf1) with a simultaneous decrease in cdk2, cdk4, cyclin D1 and cyclin E expression. The induction of apoptosis after treatment with ApoG2 for 12, 24 and 48 h was demonstrated by flow cytometry analysis. ApoG2 also induced cytochrome c release and activation of caspase-3. To our knowledge, this is the first time that ApoG2 has been reported to potently inhibit the proliferation of human monocytic lymphoma U937 cells through G1 arrest. These findings suggest that ApoG2 may be a potential chemotherapeutic agent for the treatment of cancer.

Topics: Antineoplastic Agents; Apoptosis; Caspase 3; Cell Cycle; Cell Proliferation; Cell Survival; Cyclin D1; Cyclin E; Cyclin-Dependent Kinase 2; Cyclin-Dependent Kinase 4; Cyclin-Dependent Kinase Inhibitor p21; Cytochromes c; Dose-Response Relationship, Drug; Gossypol; Humans; Lymphoma; Mitochondria; Proto-Oncogene Proteins c-myc; Time Factors; U937 Cells

Cyclin D1-negative mantle cell lymphoma is difficult to distinguish from other small B-cell lymphomas. The clinical and pathological characteristics of patients with this form of lymphoma have not been well defined. Overexpression of the transcription factor SOX11 has been observed in conventional mantle cell lymphoma. The aim of this study was to determine whether this gene is expressed in cyclin D1-negative mantle cell lymphoma and whether its detection may be useful to identify these tumors.. The microarray database of 238 mature B-cell neoplasms was re-examined. SOX11 protein expression was investigated immunohistochemically in 12 cases of cyclin D1-negative mantle cell lymphoma, 54 cases of conventional mantle cell lymphoma, and 209 additional lymphoid neoplasms.. SOX11 mRNA was highly expressed in conventional and cyclin D1-negative mantle cell lymphoma and in 33% of the cases of Burkitt's lymphoma but not in any other mature lymphoid neoplasm. SOX11 nuclear protein was detected in 50 cases (93%) of conventional mantle cell lymphoma and also in the 12 cyclin D1-negative cases of mantle cell lymphoma, the six cases of lymphoblastic lymphomas, in two of eight cases of Burkitt's lymphoma, and in two of three T-prolymphocytic leukemias but was negative in the remaining lymphoid neoplasms. Cyclin D2 and D3 mRNA levels were significantly higher in cyclin D1-negative mantle cell lymphoma than in conventional mantle cell lymphoma but the protein expression was not discriminative. The clinico-pathological features and outcomes of the patients with cyclin D1-negative mantle cell lymphoma identified by SOX11 expression were similar to those of patients with conventional mantle cell lymphoma.. SOX11 mRNA and nuclear protein expression is a highly specific marker for both cyclin D1-positive and negative mantle cell lymphoma.

Topics: Adult; Aged; Biomarkers, Tumor; Cyclin D1; Female; Gene Expression Regulation, Neoplastic; Humans; Lymphoma; Lymphoma, Mantle-Cell; Male; Middle Aged; Neoplasm Proteins; RNA, Messenger; SOXC Transcription Factors; Treatment Outcome

Clonally related composite lymphomas of Hodgkin's lymphoma (HL) and Non-Hodgkin's lymphoma (NHL) represent models to study the multistep transformation process in tumorigenesis and the development of two distinct tumors from a shared precursor. We analyzed six such lymphomas for transforming events. The HLs were combined in two cases with follicular lymphoma (FL), and in one case each with B-cell chronic lymphocytic leukemia, splenic marginal zone lymphoma, mantle cell lymphoma (MCL) and diffuse large B-cell lymphoma (DLBCL). In the HL/FL and HL/MCL combinations, BCL2/IGH and CCND1/IGH translocations, respectively, were detected in both the HL and NHL. No mutations were found in the tumor suppressor genes FAS, NFKBIA and ATM. The HL/DLBCL case harbored clonal replacement mutations of the TP53 gene on both alleles exclusively in the DLBCL. In conclusion, we present the first examples of molecularly verified IgH-associated translocations in HL, which also show that BCL2/IGH or CCND1/IGH translocations can represent early steps in the pathogenesis of composite HL/FL or HL/MCL. The restriction of the TP53 mutations to the DLBCL in the HL/DLBCL case exemplifies a late transforming event that presumably happened in the germinal center and affected the fate of a common lymphoma precursor cell towards development of a DLBCL.

Topics: Cell Transformation, Neoplastic; Clone Cells; Cyclin D1; Genes, bcl-2; Genes, Tumor Suppressor; Hodgkin Disease; Humans; Immunoglobulin Heavy Chains; Lymphoma; Lymphoma, Non-Hodgkin; Mutation; Translocation, Genetic; Tumor Suppressor Protein p53

Cyclin D1 expression is deregulated by chromosome translocation in mantle cell lymphoma and a subset of multiple myeloma. The molecular mechanisms involved in long-distance gene deregulation remain obscure, although changes in acetylated histones and methylated CpG dinucleotides may be important. The patterns of DNA methylation and histone acetylation were determined at the cyclin D1 locus on chromosome 11q13 in B-cell malignancies. The cyclin D1 promoter was hypomethylated and hyperacetylated in expressing cell lines and patient samples, and methylated and hypoacetylated in nonexpressing cell lines. Domains of hyperacetylated histones and hypomethylated DNA extended over 120 kb upstream of the cyclin D1 gene. Interestingly, hypomethylated DNA and hyperacetylated histones were also located at the cyclin D1 promoter but not the upstream major translocation cluster region in cyclin D1-nonexpressing, nontumorigenic B and T cells. RNA polymerase II binding was demonstrated both at the cyclin D1 promoter and 3' immunoglobulin heavy-chain regulatory regions only in malignant B-cell lines with deregulated cyclin D1 expression. Our results suggest a model where RNA polymerase II bound at IgH regulatory sequences can activate the cyclin D1 promoter by either long-range polymerase transfer or tracking.

Topics: Acetylation; B-Lymphocytes; Cell Line, Tumor; Chromosomes, Human, Pair 11; Chromosomes, Human, Pair 14; Cyclin D1; DNA; DNA Methylation; Gene Expression; Histones; Humans; Immunoglobulin Heavy Chains; Lymphoma; Promoter Regions, Genetic; Protein Binding; RNA Polymerase II; T-Lymphocytes

Signal cascades of mitogen-actived protein kinase(MAPK) and signal transducer and activator of transcription 3 (Stat3) are two main signal transduction pathways which were associated with cell proliferation and malignant transformation.MAPK and Stat3 proteins are activated by phosphorylation. The biological effects which are caused by multiple cytokines produced by Hodgkin's lymphoma(HL) cells are mediated through MAPK and Stat3 signal pathways. This study was designed to investigate significance of MAPK and Stat3 phosphorylation(p-MAPK and p-Stat3) and cyclin D1 protein expression in HL.. SP immunohistochemistry was used to detect expression of p-MAPK, p-Stat3, and cyclin D1 protein in 45 cases of HL of various types.. The expression positive rates of p-MAPK, p-Stat3, and cyclin D1 proteins were 73.3%(33/45),64.4%(29/45), and 68.9%(31/45), respectively. The positive expression levels of p-MAPK and cyclin D1 protein gradually increased(P< 0.05), whereas that of p-stat3 had no significant difference(P >0.05) in four subsets(LR:lymphocyte-rich classical type; NS:nodular sclerosis type; MC:mixed cellularity type; LD:lymphocyte depletion type) of all cases. The expression of p-MAPK was positively related to that of cyclin D1 protein (r(s)=0.7254,P< 0.01), but the expression of p-Stat3 was not related to that of cyclin D1 protein (r(s)=0.2197,P >0.05).. The data suggest that activation of MAPK may play an important role in genesis and progression of HL, but Stat3 activation is not associated with the progression of HL. MAPK may induce overexpression of cyclin D1 protein and results in persistent proliferation of RS/H cells in genesis and development of HL.

Topics: Adolescent; Adult; Aged; Child; Child, Preschool; Cyclin D1; DNA-Binding Proteins; Female; Hodgkin Disease; Humans; Immunohistochemistry; Lymphoma; Male; MAP Kinase Signaling System; Middle Aged; Mitogen-Activated Protein Kinases; Phosphorylation; Signal Transduction; STAT3 Transcription Factor; Trans-Activators; Transcription, Genetic

Topics: Animals; Annexin A5; Antineoplastic Combined Chemotherapy Protocols; Cyclin D1; Genes, bcl-1; Genes, Reporter; Humans; Luciferases; Lung Neoplasms; Lymphoma; Mice; Mice, Inbred BALB C; Organotechnetium Compounds; Rabbits; Radionuclide Imaging; Radiopharmaceuticals; Recombinant Fusion Proteins; Time Factors

Most lymphoid malignancies are initiated by specific chromosomal translocations between immunoglobulin (Ig)/T cell receptor (TCR) gene segments and cellular proto-oncogenes. In many cases, illegitimate V(D)J recombination has been proposed to be involved in the translocation process, but this has never been functionally established. Using extra-chromosomal recombination assays, we determined the ability of several proto-oncogenes to target V(D)J recombination, and assessed the impact of their recombinogenic potential on translocation rates in vivo. Our data support the involvement of 2 distinct mechanisms: translocations involving LMO2, TAL2, and TAL1 in T cell acute lymphoblastic leukemia (T-ALL), are compatible with illegitimate V(D)J recombination between a TCR locus and a proto-oncogene locus bearing a fortuitous but functional recombination site (type 1); in contrast, translocations involving BCL1 and BCL2 in B cell non-Hodgkin's lymphomas (B-NHL), are compatible with a process in which only the IgH locus breaks are mediated by V(D)J recombination (type 2). Most importantly, we show that the t(11;14)(p13;q32) translocation involving LMO2 is present at strikingly high frequency in normal human thymus, and that the recombinogenic potential conferred by the LMO2 cryptic site is directly predictive of the in vivo level of translocation at that locus. These findings provide new insights into the regulation forces acting upon genomic instability in B and T cell tumorigenesis.

Topics: Adaptor Proteins, Signal Transducing; Animals; Basic Helix-Loop-Helix Transcription Factors; Child; Cyclin D1; DNA-Binding Proteins; Humans; Leukemia-Lymphoma, Adult T-Cell; Leukemia, Lymphoid; LIM Domain Proteins; Lymphoma; Metalloproteins; Mice; Models, Genetic; Neoplasm Proteins; Proto-Oncogene Mas; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-bcl-2; Recombination, Genetic; T-Cell Acute Lymphocytic Leukemia Protein 1; Thymus Gland; Transcription Factors; Translocation, Genetic

We have used a continuous fluorescence monitoring method to assess cyclin D1 mRNA expression in a variety of hematological and non-hematological processes. We examined 14 cell lines, 11 reactive lymphoid tissues, and 57 primary hematopoietic neoplasms including mantle cell lymphoma (MCL) (n = 10), chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL) (n = 11), acute lymphoblastic leukemia/lymphoma (n = 15), follicular lymphoma (n = 6), peripheral T-cell lymphoma (PTCL) (n = 3), anaplastic large cell lymphoma (n = 3), hairy cell leukemia (n = 3), Burkitt lymphoma (n = 1), Burkitt-like lymphoma (n = 4), and plasmacytoma (n = 1) for the expression of cyclin D1 mRNA using fluorescently labeled sequence-specific hybridization probes. Fluorescence (F) was plotted against cycle (C) number over 45 cycles. The log-linear portion of the F versus C graph identified a fractional cycle number for threshold fluorescence. A beta-globin mRNA transcript with equivalent amplification efficiency to that of cyclin D1 was used for assessment of RNA integrity and normalization. In general, the MCLs demonstrated substantially higher levels of cyclin D1 mRNA than the other lymphoproliferative processes. Moderately high levels of cyclin D1 mRNA were detected in one PTCL. On average, the CLL/SLL cases showed cyclin D1 mRNA levels two to three orders of magnitude lower than observed in the MCLs. Cell lines derived from non-hematopoietic neoplasms such as fibrosarcoma, small cell carcinoma, and neuroblastoma showed comparable or higher levels of cyclin D1 mRNA than the MCLs. Our results indicate that quantitative real-time reverse transcription (RT) polymerase chain reaction is a simple, rapid, and accurate technique for assessing cyclin D1 expression, and while it is not specific, it can reliably be used in the distinction of MCL from CLL/SLL.

Topics: Beta-Globulins; Biomarkers, Tumor; Cyclin D1; DNA Primers; Fluorescence; Humans; Immunoenzyme Techniques; Lymphoma; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; RNA, Neoplasm; Tumor Cells, Cultured

The pRb pathway plays a key role in controlling the G1/S transition in cell cycle progression. Aberrations of various components of the pRb pathway, such as retinoblastoma protein and its upstream actors including cyclin D1, cyclin dependence kinase-4 and p16/p15 cyclin dependent kinase inhibitors, have been reported in a variety of human tumors. Furthermore, the alterations of retinoblastoma protein and its upstream components often occur in a reciprocal manner. Previously, we have reported frequent inactivation of the Cdkn2a/Cdkn2b loci encoding p16/p15 cyclin dependent kinase inhibitors in a subset of 2',3'-dideoxycytidine- and 1, 3-butadiene-induced mouse lymphomas (S.-M. Zhuang, A. Schippert, A. Haugen-Strano, R.W. Wiseman, P. Söderkvist, Inactivation of p16(INK4a)-alpha, p16(INK4a)-beta and p15(INK4b) genes in 2', 3'-dideoxycytidine- and 1,3-butadiene-induced lymphomas, Oncogene 16 (1998) 803-808), indicating the involvement of pRb pathway in lymphomagenesis. To investigate whether alteration of other components in pRb pathway is an alternative mechanism underlying the development of these chemically induced lymphomas, we have examined the genetic status of Rb1, Ccnd1 and Cdk4 genes that encode retinoblastoma protein, cyclin D1 and cyclin dependence kinase-4, respectively. Gross alterations of the Rb1, Ccnd1, and Cdk4 genes were not detected by Southern analysis in any of the tumors examined. In addition, single-strand conformation analysis failed to reveal point mutations in the Cdk4 amino terminal domain that is important for its association with Cdkn2a gene products. These results indicate that the mechanisms underlying the development of 2', 3'-dideoxycytidine- and 1,3-butadiene-induced lymphomas involve inactivation of p16/p15 cyclin-dependent kinase inhibitors but not genomic alterations of the Rb1, Ccnd1 and Cdk4 genes.

Topics: Animals; Blotting, Southern; Butadienes; Carcinogens; Cyclin D1; Cyclin-Dependent Kinase 4; Cyclin-Dependent Kinases; DNA Mutational Analysis; Genes, Retinoblastoma; Lymphoma; Mice; Mice, Inbred C3H; Mice, Inbred C57BL; Polymorphism, Single-Stranded Conformational; Proto-Oncogene Proteins; Zalcitabine

Conventional chromosome analysis (CCA) and interphase fluorescence in situ hybridization (FISH) was performed in 42 patients with mantle-cell lymphoma (MCL), with BCL1 rearrangement. The t(11;14)(q13;q32) or 11q abnormalities were detected by CCA in 34 cases, 20 of which had additional aberrations. A normal karyotype was observed in 8 cases. Probes detecting the chromosome aberrations that were observed in at least 3 cases by CCA, ie, +12, 13q14 deletion, and 17p deletion, were used for interphase FISH analysis. FISH detected total or partial +12, 13q14 deletion and 17p- in 28.5%, 52.4%, and 26% of the cases, respectively. The presence of these anomalies was not a function of karyotype complexity. Based on the results of CCA/FISH, three groups of increasing karyotype complexity were recognized: group 1, including 11 patients without detectable aberrations in addition to BCL1 rearrangement; group 2, including 14 patients with 1 to 2 additional anomalies; and group 3, including 17 patients with three or more additional anomalies. Clinical parameters associated with shorter survival were male sex (P =.006) and primary lymph-node involvement compared with primary bone marrow involvement (P =.015). Trisomy 12 was the only single cytogenetic parameter predictive of a poor prognosis (P =.006) and the best prognostic indicator was the derived measure of karyotype complexity (P <.0001), which maintained statistical significance in multivariate analysis (P<.0001). We arrived at the following conclusions: 13q14 deletion occurs at a high incidence in MCL; 17p deletion and total/partial +12 are relatively frequent events in MCL, the latter aberration being associated with a shorter survival; and the degree of karyotype complexity has a strong impact on prognosis in this neoplasia.

Topics: Adult; Aged; Aged, 80 and over; Cell Lineage; Chromosomes, Human, Pair 11; Chromosomes, Human, Pair 14; Cyclin D1; Female; Gene Rearrangement; Humans; Lymphoma; Male; Middle Aged; Translocation, Genetic

We observed the expression of wild-type retinoblastoma protein (RB) in all 17 hematologic cultured cell lines tested. However, no p16INK4 expression was detected in any cell line among 16 leukemia/lymphoma cell lines, although an EBV-transformed cell line expressed p16INK4. The expression levels of cyclin D1 and CDK4 varied widely among the cell lines. The correlation coefficient (r2) between doubling time (DT) and cyclin D1 in the 14 cell lines that doubled within 47.2 h was 0.4856, while the r2 between DT and cyclin dependent kinase 4 (CDK4) and that between DT and RB among those cell lines were 0.3761 and 0.0874, respectively. The levels of protein expression in vincristine (VCR)-resistant cell lines was not different from those in corresponding wild-type cell lines. Thus, we concluded that the loss of p16INK4 protein and inactivation of RB protein could be an essential step for oncogenesis of leukemia/ lymphoma, and that cyclin D1 may possibly be a target protein to control cell growth of hematologic cell lines which lack the expression of p16INK4.

Topics: Cell Division; Cyclin D1; Cyclin-Dependent Kinase 4; Cyclin-Dependent Kinase Inhibitor p16; Cyclin-Dependent Kinases; G1 Phase; Humans; Leukemia; Lymphoma; Proto-Oncogene Proteins; Retinoblastoma Protein; Tumor Cells, Cultured

To investigate the regulatory mechanism of the expression of the multidrug resistance gene (mdr-1) and the multidrug resistance-associated protein gene (mrp), we investigated if p53, WT1, RB, C-myc, N-myc, cyclin D1, p16INK4 (p16) are involved in the acquirement of multidrug resistance phenotype (MDR) in human vincristine (VCR)-resistant cells of leukemia/lymphoma cell lines. By using RT-PCR, we observed that MDR in VCR-resistant cell lines was mediated by either mdr-1 or mrp genes. In cells that acquired the overexpression of mdr-1, downregulation of p53 and upregulation of WT-1 were observed. In contrast, no constant change of genes was observed in cells that overexpressed mrp. Although the change in the expression level of cyclin D1 and p-16 accompanied the development of VCR resistance, the mRNA of RB, C-myc and N-myc showed no correlation with the degree of VCR resistance or the level of mdr-1 expression. These results may provide a plausible diagnostic marker for determination of drug sensitivity in cancer patients and suggest that p53 may mediate directly or indirectly the expression of mdr-1 via WT1 in VCR-resistant hematologic cell lines.

Topics: ATP Binding Cassette Transporter, Subfamily B, Member 1; ATP-Binding Cassette Transporters; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Cytarabine; DNA-Binding Proteins; Doxorubicin; Drug Resistance, Neoplasm; Gene Expression Regulation, Neoplastic; Genes, p53; Genes, Retinoblastoma; Humans; Leukemia; Lymphoma; Multidrug Resistance-Associated Proteins; Proto-Oncogene Proteins c-myc; RNA, Messenger; RNA, Neoplasm; Transcription Factors; Tumor Cells, Cultured; Vincristine; WT1 Proteins

To evaluate the utility of a polymerase chain reaction (PCR)-based, B-cell-related molecular panel in the diagnostic workup of hematologic specimens.. A PCR panel was applied to 89 specimens from 87 patients, including 55 cases (57 specimens) of known monoclonal B-cell lymphoma, 10 cases of Hodgkin's disease, 2 cases of T-cell lymphoma, and 20 specimens of polyclonal reactive lymphoid tissues. The panel comprised a seminested PCR procedure for immunoglobulin heavy-chain (IgH) gene rearrangement and unnested PCR detection of t(14;18) and t(11;14).. Immunoglobulin heavy-chain was detected in 63%, evidence of t(14;18) in 35%, and evidence of t(11;14) in 3.5% of all the B-cell lymphoma cases. Seventy-seven percent of all cases demonstrated IgH- and/or bcl-2-associated translocations using these primers. The IgH primers alone detected clonality in 82% (28/34) of the nonfollicular lymphomas and 35% (8/23) of the follicular lymphomas, with no false positives in the non-B-cell lymphoma specimens. The addition of two primer sets for the detection of both IgH and bcl-2/JH significantly improved the detection of molecular abnormalities in the follicular lymphoma group from 35% to 70% (16/23). However, no change was noted in the overall detection rate for the nonfollicular lymphoma group. Adding primers for bcl-1/JH did not change the overall detection rate for either group.. Seminested PCR for IgH detected monoclonality in the majority of the nonfollicular lymphomas and is a valuable diagnostic tool in this group. The combination of different primer sets for the detection of IgH gene rearrangement and bcl-2/JH is most desirable for follicular lymphomas.

Topics: Base Sequence; Biomarkers, Tumor; Cyclin D1; Cyclins; DNA Primers; DNA, Neoplasm; Electrophoresis, Agar Gel; Gene Rearrangement; Genes, Immunoglobulin; Humans; Immunoglobulin Heavy Chains; Lymphoma; Lymphoma, B-Cell; Molecular Sequence Data; Oncogene Proteins; Polymerase Chain Reaction; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-bcl-2; Retrospective Studies; Translocation, Genetic

Aberrations of the long arm of chromosome 11 are among the most common chromosome abnormalities in lymphoproliferative disorders (LPD). Translocations involving BCL1 at 11q13 are strongly associated with mantle cell lymphoma. other nonrandom aberrations, especially deletions and, less frequently, translocations, involving bands 11q21-923 have been identified by chromosome banding analysis. To date, the critical genomic segment and candidate genes involved in these deletions have not been identified. In the present study, we have analyzed tumors from 43 patients with LPD (B-cell chronic lymphocytic leukemia, n = 40; mantle cell lymphoma, n = 3) showing aberrations of bands 11q21-923 by fluorescence in situ hybridization. As probes we used Alu-PCR products from 17 yeast artificial chromosome clones spanning chromosome bands 11q14.3-923.3, including a panel of yeast artificial chromosome clones recognizing a contiguous genomic DNA fragment of approximately 9-10 Mb in bands 11q22.3-923.3. In the 41 tumors exhibiting deletions, we identified a commonly deleted segment in band 11q22.3-923.1; this region is approximately 2-3 Mb in size and contains the genes coding for ATM (ataxia telangiectasia mutated), RDX (radixin), and FDX1 (ferredoxin 1). Furthermore, two translocation break-points were localized to a 1.8-Mb genomic fragment contained within the commonly deleted segment. Thus, we have identified a single critical region of 2-3 Mb in size in which 11q14-923 aberrations in LPD cluster. This provides the basis for the identification of the gene(s) at 11q22.3-923.1 that are involved in the pathogenesis of LPD.

Topics: Ataxia Telangiectasia Mutated Proteins; Blood Proteins; Cell Cycle Proteins; Chromosome Aberrations; Chromosome Banding; Chromosome Disorders; Chromosome Mapping; Chromosomes, Artificial, Yeast; Chromosomes, Human, Pair 11; Cyclin D1; Cytoskeletal Proteins; DNA-Binding Proteins; Ferredoxins; Gene Deletion; Humans; In Situ Hybridization, Fluorescence; Lymphoma; Lymphoproliferative Disorders; Membrane Proteins; Protein Serine-Threonine Kinases; Proteins; Proto-Oncogene Proteins; Tumor Suppressor Proteins

Cyclin D1/PRAD1 (bcl-1) is a recently discovered proto-oncogene that is overexpressed in mantle cell lymphomas and several other human tumors. In a previous study, the authors demonstrated expression of cyclin D1 in 15 of 15 cases of mantle cell lymphoma and 1 of 8 cases of B-chronic lymphocytic leukemia (B-CLL) using a polyclonal antibody and microwave enhanced immunohistochemical staining method on paraffin-embedded tissue sections. In this study, 107 additional B- and T-cell neoplasms were studied, including 47 cases of high grade lymphoma (33 diffuse large B-cell type, 9 Burkitt and Burkitt-like, 4 precursor T-lymphoblastic lymphoma, and 1 adult T-cell lymphoma/leukemia), 38 additional cases of low grade B-cell lymphoma (18 CLL, 15 hairy cell leukemia and 5 mantle cell lymphoma), and 22 plasmacytomas for expression of cyclin D1 using the same immunohistochemical staining technique. All cases of mantle cell lymphoma showed diffuse nuclear staining. No additional cases of CLL showed cyclin D1 expression. In contrast, 1 of 15 hairy cell leukemias and 1 of 22 plasmacytomas showed cyclin D1 staining. None of the high grade lymphomas demonstrated expression of cyclin D1 protein by immunostaining, including three cases of large B-cell lymphoma that coexpressed CD5. The authors conclude that cyclin D1 is expressed in all cases of mantle cell lymphoma, and only in very rare cases of B-CLL, hairy cell leukemia and plasmacytoma/myeloma. Cyclin D1 does not appear to play an important role in high grade lymphomas. In addition, most CD5 positive high grade B-cell lymphomas do not express cyclin D1, and are not likely to be derived from mantle cell lymphoma or other lymphomas with t(11;14).

Topics: Cyclin D1; Cyclins; Humans; Immunohistochemistry; Leukemia, Hairy Cell; Lymphoma; Lymphoma, B-Cell; Lymphoma, Non-Hodgkin; Multiple Myeloma; Oncogene Proteins; Plasmacytoma; Proto-Oncogene Mas; Staining and Labeling

The t(11;14)(q13;q32) translocation and its molecular counterpart bcl-1 rearrangement are frequently associated with mantle cell lymphomas (MCLs) and only occasionally with other variants of B-cell lymphoid malignancies. This translocation seems to activate the expression of PRAD-1/cyclin D1 gene located downstream from the major breakpoint cluster region of this rearrangement. However, the possible overexpression of this gene in other lymphoproliferative disorders independently of bcl-1 rearrangement is unknown. We have examined the overexpression of PRAD-1 gene in a large series of 142 lymphoproliferative disorders including 20 MCLs by Northern blot analysis. Cytogenetic and/or bcl-1 rearrangement analysis with 2 probes (MTC, p94PS) were performed in 28 cases. Strong PRAD-1 overexpression was observed in 19 of the 20 MCLs including 3 gastrointestinal forms and 4 blastic variants. t(11;14) and/or bcl-1 rearrangement was detected in 6 of the 12 MCLs examined. No correlation was found between the different levels of mRNA expression and the pathologic characteristics of the lymphoma. Among chronic lymphoproliferative disorders other than MCL, only 1 atypical chronic lymphocytic leukemia (CLL) with a t(11;14) translocation and bcl-1 rearrangement and the 2 hairy cell leukemias (HCLs) analyzed showed upregulation of PRAD-1 gene. The expression in the 2 HCLs was lower than in MCL, and no bcl-1 rearrangement was observed. These findings indicate that PRAD-1 overexpression is a highly sensitive and specific molecular marker of MCL but it may also be upregulated in some B-CLLs and in HCL.

Topics: Biomarkers, Tumor; Blotting, Northern; Chronic Disease; Cyclin D1; Cyclins; Gene Expression; Gene Rearrangement; Humans; Leukemia, Hairy Cell; Leukemia, Lymphocytic, Chronic, B-Cell; Lymphoma; Lymphoproliferative Disorders; Oncogene Proteins; Proto-Oncogene Proteins; RNA, Messenger; Translocation, Genetic

Mouse monoclonal antibodies were produced against the bacterial product encoded by human PRAD1/cyclin D1 gene, which is known to be involved in tumors with translocation or amplification at BCL-1 locus of 11q13. The immunizing antigens used were GST-PRAD1 and T7 gene 10-PRAD1 fusion products. Four antibodies were reactive with both PRAD1 fusion products and cell lysates of B-cell tumor cell lines with t(11;14)(q13;q32) and a breast cancer cell line with 11q13 amplification, on immunoblotting. An immunofluorescence study showed that only one of them stained nuclei of cells with 11q13 abnormalities. Since this antibody proved applicable for conventional paraffin-embedded tissue sections, immunohistologic staining of various lymphoma tissues was performed. Eight of 11 mantle cell lymphomas showed intermediate to strong positivity and 6 of the positive cases demonstrated characteristic staining patterns that were either predominantly nuclear or both nuclear and cytoplasmic. The nuclear staining pattern was not observed with other types of lymphoma and thus may correlate with PRAD1 mRNA overexpression.

Topics: Antibodies, Monoclonal; Cell Nucleus; Chromosome Aberrations; Chromosome Disorders; Chromosomes, Human, Pair 11; Chromosomes, Human, Pair 14; Cyclin D1; Cyclins; DNA, Neoplasm; Gene Amplification; Gene Expression Regulation, Neoplastic; Genes; Humans; Lymphoma; Oncogene Proteins; Proto-Oncogene Proteins; Recombinant Fusion Proteins; RNA, Messenger; RNA, Neoplasm; Translocation, Genetic

The 11q13 breakpoint region of t(11;14) (q13;q32), translocated to the Ig heavy chain locus at 14q32, has been designated as BCL-1 for B-cell leukemia/lymphoma-1, but the nature of the transcriptional unit has long remained unclear. Recently, the PRAD1 gene encoding cyclin D1, isolated from the 11q13 region, was proposed as a candidate BCL-1 gene on the basis of chromosome walking and concordant overexpression of PRAD1 mRNA in cell lines with t(11;14)(q13;q32). We report here molecular analysis of a variant translocation at the BCL-1 locus, t(11;22)(q13;q11), showing juxtaposition of the Ig light chain gene, Ig lambda, to the PRAD1 gene at its 3' end, resulting in overexpression of PRAD1 mRNA. Because only the PRAD1 gene is present between the Ig heavy chain and light chain gene breakpoints, an identity between BCL-1 and the PRAD1/cyclin D1 gene is strongly indicated.

Topics: Base Sequence; Chromosome Mapping; Chromosome Walking; Chromosomes, Human, Pair 11; Chromosomes, Human, Pair 14; Chromosomes, Human, Pair 22; Cyclin D1; Cyclins; DNA Primers; Gene Expression; Genetic Variation; Humans; Immunoglobulin Heavy Chains; Immunoglobulin lambda-Chains; Immunoglobulin Light Chains; Introns; Leukemia, Lymphocytic, Chronic, B-Cell; Lymphoma; Molecular Sequence Data; Oncogene Proteins; Polymerase Chain Reaction; RNA, Messenger; Translocation, Genetic

The translocation t(11;14)(q13;q32) occurs in about 15% of patients with splenic lymphoma with villous lymphocytes (SLVL) or the closely related disorder lymphoplasmacytic lymphoma (LPL). To characterize the nature and frequency of rearrangements of the BCL-1 locus in SLVL/LPL and to document the effect of these genetic alterations on the expression of the cyclin D1 gene, we analyzed 22 cases of SLVL/LPL with defined cytogenetic abnormalities by both conventional electrophoresis (CE) and pulse-field gel electrophoresis (PFGE) and by Northern blotting. Four SLVL/LPL cases showed rearrangement of the BCL-1 locus. In two cases with t(11;14)(q13;q32), different breakpoints were identified; one mapped adjacent to the major translocation cluster (MTC) and the other within a 28-kb region telomeric of it. In a third case of SLVL with no cytogenetic abnormality of 11q13, a novel breakpoint approximately 100 kb centromeric of MTC was detected by PFGE. The fourth case, which had a normal karyotype, demonstrated rearrangement with a BCL-1 probe immediately telomeric of MTC. This case may have had a small deletion of 0.5 kb from within the BCL-1 locus. No rearrangement of the BCL-1 locus or within the cyclin D1 gene was detected by CE or PFGE in any of the remaining 18 SLVL/LPL samples. Northern blot analysis showed expression of a normal-sized cyclin D1 transcript in the two SLVL/LPL cases with t(11;14)(q13;q32). In cases that lacked a cytogenetically demonstrable t(11;14) translocation, no cyclin D1 transcript was detected. Analysis of the BCL-1 locus was also performed in three other cases of B-cell disorders with t(11;14)(q13;q32) detected cytogenetically. Two cases were analyzed by Southern blot and showed rearrangement of the BCL-1 locus. Expression of high-level normal-sized and/or truncated cyclin D1 transcript was also detected in these cases. These data show the importance of PFGE in the detection of rearrangements in the BCL-1 locus and show further the complexity of rearrangements in this locus.

Topics: Chromosomes, Human, Pair 11; Chromosomes, Human, Pair 14; Cyclin D1; Cyclins; Gene Rearrangement; Genes, Immunoglobulin; Humans; Immunoglobulin Heavy Chains; Lymphocytes; Lymphoma; Oncogene Proteins; Proto-Oncogene Proteins; Proto-Oncogenes; Splenic Neoplasms; Translocation, Genetic

PRAD1 (cyclin D1) is a recently identified member of the family of cyclin genes, believed to play roles in regulating transitions through the cell cycle. The PRAD1 gene, located at 11q13, has been implicated in the pathogenesis of a variety of tumors, including parathyroid adenomas, t(11;14) bearing B-lymphoid tumors (particularly centrocytic lymphomas) where it is highly likely to be the BCL1 oncogene, and possibly in breast carcinomas and squamous cell cancers of the head and neck as well. PRAD1's tumorigenic influence appears to be effected through overexpression of its normal-sized transcript, but it has not been established whether the transcript's coding sequence is normal or contains oncogenic mutations. We have sequenced the coding region of the overexpressed PRAD1 transcript from two primary tumors with clonal PRAD1 region rearrangements: a benign parathyroid adenoma and a malignant centrocytic lymphoma. Each sequence is identical to the normal PRAD1 cDNA sequence, and presumably encodes normal PRAD1 protein. Thus, PRAD1 likely functions as a direct-acting oncogene whose rearrangement in tumors leads to overexpression or deregulated expression of its normal protein product.

Topics: Adenoma; Base Sequence; Cyclin D1; Cyclins; Gene Expression; Gene Rearrangement; Humans; Lymphoma; Molecular Sequence Data; Oncogene Proteins; Oncogenes; Parathyroid Neoplasms

By isolating genomic DNA clones that encompass the mouse Cyl-1 (cyclin D1) locus, we have identified a putative CpG island close to the 5' end of the gene. Pulsed-field gel electrophoresis with probes derived from either the 5' or 3' side of the CpG island established physical linkage to two independent markers on mouse chromosome 7, in a region that is syntenic with human chromosome 11q13. On the 3' side, Cyl-1 is approximately 75 kb from Hst-1 and Int-2, although there is an additional CpG island in the intervening DNA, while on the 5' side, Cyl-1 is less than 300 kb from Fis-1, an integration site for Friend murine leukaemia virus. As there is no intervening CpG island, proviral insertions at Fis-1 could influence the expression of Cyl-1 and we describe two virally induced tumours in which this appears to be the case. The data suggest that proviral insertions near Cyl-1 in mouse lymphomas are functionally equivalent to the BCL1 translocations that activate cyclin D1 expression in human B-cell malignancies.

Topics: Animals; Blotting, Southern; Chromosome Mapping; Chromosomes, Human, Pair 11; Cosmids; Cyclin D1; Cyclins; DNA; DNA, Neoplasm; Friend murine leukemia virus; Gene Library; Genetic Linkage; Humans; Lymphoma; Lymphoma, B-Cell; Mice; Mice, Inbred BALB C; Oncogene Proteins; Oncogenes; Proviruses; Restriction Mapping; Translocation, Genetic

The organization of immunoglobulin heavy chain (IgH), light chain (kappa and lambda) and T cell receptor (TCR) beta chain gene loci in 10 patients with ocular adnexal pseudolymphoma was investigated. Eight of them showed IgH gene rearrangement in at least one of the 3 restriction enzymes-digested DNAs extracted from ocular adnexal neoplasms. In contrast, none of them exhibited clonal TCR beta chain gene rearrangement. The configuration of bcl-1, bcl-2 and c-myc oncogenes was also studied by Southern blot technique. Two patients had a rearranged joining region, IgH-containing fragment that comigrated with the rearranged bcl-1 fragment. C-myc gene rearrangement was found in only one patient who also had bcl-1 gene rearrangement. In ocular adnexal pseudolymphoma, none demonstrated bcl-2 gene rearrangement; however, in bone marrow cells, one patient with systemic lymphadenopathy exhibited both IgH and bcl-2 gene rearrangements. Three genotypic subsets of these ocular adnexal pseudolymphoma can be identified by the configuration of IgH gene and related oncogenes: with germline configuration of IgH gene and bcl-1, bcl-2 and c-myc oncogenes; with rearrangement of IgH gene but germline configuration of these oncogenes; and with recombination between rearranged IgH and bcl-1 genes. These results suggest in ocular adnexal pseudolymphoma a spectrum of clonal change evolving from polyclonal to monoclonal B-population, and further to monoclonal B-population with rearranged bcl-1, c-myc and/or bcl-2 oncogenes.

Topics: Adult; Aged; Aged, 80 and over; Cyclin D1; DNA, Neoplasm; Eyelid Neoplasms; Gene Rearrangement; Genes, Immunoglobulin; Genes, myc; Humans; Lymphoma; Male; Middle Aged; Orbital Neoplasms; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-bcl-2; Receptors, Antigen, T-Cell; Receptors, Antigen, T-Cell, alpha-beta