transforming-growth-factor-beta and Lymphoma--Large-B-Cell--Diffuse

The development of drug resistance remains the major obstacle to clinical efficacy of cancer chemotherapy. Consequently, finding new therapeutic options for cancerous patients is an urgent need. Sixty newly diagnosed diffuse large B-cell lymphoma (DLBCL) patients were recruited from Clinical Oncology Department, Faculty of Medicine, Menoufia University, Egypt prospectively randomized to three groups (n = 20 for each group). Group one (control group) received R-CHOP standard chemotherapy {Rituximab, Cyclophosphamide, Hydroxyldaunorubicin (Doxorubicin)®, Vincristine (oncovin)®, prednisolone in the first five days of cycle}, group two received lansoprazole (LAN) 60 mg p.o. bid for only one week before starting each of cycle + R-CHOP and group three received famotidine (FAM) 40 mg p.o. once daily one week before cycle and continues daily through the cycle + R-CHOP for six cycles. Blood samples were obtained for biochemical analysis of transforming growth factor-β (TGF-β), Basic fibroblast growth factor (bFGF), interleukin-9 (IL-9), nuclear factor-kappa B (NF-κB) and Caspase 3 before and after six cycles of therapy. The obtained data showed that LAN and FAM resulted in significant decrease in (LDH, TGF-β, bFGF and IL-9, respectively) and significant increase in (Caspase-3). In addition, LAN produced a significant elevation in the response rate compared to the control group or the FAM group. Both LAN and FAM as adjuvant therapy represents a promising anticancer strategy in DLBCL by modulation of malignancy homeostasis mechanisms and boosting chemotherapy antitumor effects without further toxicity. In addition, LAN has a synergetic effect in improving the response rate.Trial registration Clinical Trial.gov Identifier: NCT0364707.

Topics: Adult; Antineoplastic Combined Chemotherapy Protocols; Caspase 3; Cyclophosphamide; Doxorubicin; Famotidine; Female; Histamine H2 Antagonists; Humans; Lansoprazole; Lymphoma, Large B-Cell, Diffuse; Male; Middle Aged; Prednisone; Proton Pump Inhibitors; Rituximab; Transforming Growth Factor beta; Treatment Outcome; Vincristine

Based on CD25 expression, T follicular helper cells (Tfh) could be divided into T follicular regulatory (Tfr)-like subset (CD25

Topics: Adult; Aged; CD8-Positive T-Lymphocytes; Cytokines; Female; Forkhead Transcription Factors; Humans; Interleukin-10; Interleukin-2 Receptor alpha Subunit; Interleukins; Lymphoma, Large B-Cell, Diffuse; Male; Middle Aged; T Follicular Helper Cells; T-Lymphocytes, Regulatory; Transforming Growth Factor beta; Up-Regulation

Topics: Animals; Apoptosis; Cell Line, Tumor; Cell Proliferation; CRISPR-Cas Systems; Disease Models, Animal; Gene Targeting; Germinal Center; Humans; Lymphoma, Large B-Cell, Diffuse; Mice; Mice, Knockout; Models, Biological; Receptors, Lysosphingolipid; Signal Transduction; Smad1 Protein; Sphingosine-1-Phosphate Receptors; Transforming Growth Factor beta

Cancer stem cells play an important role on tumor progression. Biomarkers of stem cell property and their relationship to extranodal involvement of malignant lymphocytes are undefined in diffuse large B-cell lymphoma (DLBCL). Here we showed that junctional adhesion molecule-A (JAM-A) was highly expressed in DLBCL patients with multiple extranodal lesions. JAM-A maintained B-lymphoma cell stemness and was associated with cell invasion and epithelial-to-mesenchymal transition both in vitro and in vivo. As mechanism of action, JAM-A overexpression selectively activated transforming growth factor-β (TGF-β)/NODAL signaling, thereby enhanced B-lymphoma cell aggressiveness and induced extranodal involvement to mesoendoderm-derived organs in DLBCL. Lenalidomide downregulated JAM-A and downstream NODAL expression, resulting in inhibition of B-lymphoma cell invasion and epithelial-to-mesenchymal transition. In a murine xenograft model established with subcutaneous injection of JAM-A-overexpressing B-lymphoma cells, lenalidomide retarded tumor growth and prevented cell invasion to mesoendoderm-derived organs, consistent with the downregulation of JAM-A and NODAL expression. Collectively, these findings indicated that JAM-A was related to extranodal involvement in DLBCL through modulating TGF-β/NODAL signaling. Identified as a biomarker of stem cell property, JAM-A indicated the sensitivity of B-lymphoma cells to lenalidomide. Therapeutic targeting of JAM-A/NODAL axis could thus be a promising clinical strategy to impede tumor progression in DLBCL.

Topics: Animals; Cell Adhesion Molecules; Cell Line, Tumor; Disease Progression; Epithelial-Mesenchymal Transition; Female; Gene Expression Regulation, Neoplastic; Humans; Lenalidomide; Lymphatic Metastasis; Lymphoma, Large B-Cell, Diffuse; Male; Mice; Neoplasm Invasiveness; Nodal Protein; Prognosis; Receptors, Cell Surface; Signal Transduction; Transforming Growth Factor beta; Up-Regulation; Xenograft Model Antitumor Assays; Zebrafish

CD109 is a glycosylphosphatidylinositol-anchored glycoprotein that negatively regulates TGF-β signaling. CD109 was originally identified in hematopoietic tumors; however, the significance of CD109 in hematopoietic malignancies remains unclear. Here, we study the association of CD109 with diffuse large B-cell lymphoma (DLBCL) prognosis. Eighty-four DLBCL specimens were immunohistochemically analyzed for CD109 expression, and 31 and 53 cases were classified into low- and high-CD109 expression groups, respectively. CD109 expression was not associated with overall survival using the Kaplan-Meier analysis and log-rank tests (P = 0.17); however, a significant association was observed between high-CD109 expression and low-1-year survival (P = 0.01). Moreover, in combination with the revised International Prognostic Index (R-IPI), R-IPI-poor/CD109-high was associated with poorer prognosis compared with R-IPI-poor alone. We assessed TGF-β signaling in CD109-depleted Nalm6 cells (a human B-lymphoblastic leukemia/lymphoma cell line), and found prolonged Smad2 phosphorylation compared with control cells after TGF-β1 stimulation, suggesting that CD109 attenuates TGF-β1 signaling in human B-cell tumors. These results suggest that CD109 is a putative biomarker for identifying a high-risk group among DLBCL patients.

Topics: Adult; Aged; Aged, 80 and over; Antigens, CD; Biomarkers, Tumor; Cell Line, Tumor; Female; GPI-Linked Proteins; Humans; Lymphoma, Large B-Cell, Diffuse; Male; Middle Aged; Neoplasm Proteins; Prognosis; Risk; Signal Transduction; Transforming Growth Factor beta; Young Adult

The objective of this study was to investigate the expression and significance of the transforming growth factor β type II receptor (TGFβRII) in diffuse large B cell lymphoma. All patients were enrolled at the First Affiliated Hospital of Liaoning Medical University between 2001 and 2007. The median follow-up period was 53.3 months. Of the 338 patients studied, 131 (38.76 %) had TGFβRII positive expression on immunohistochemistry. The 5 year survival rate was significantly higher in patients with TGFβRII expression than in those without TGFβRII expression (40.3 vs. 31.6 %, P = 0.041). Multivariate analysis identified TGFβRII expression as an independent predictive parameter for survival, in addition to lactate dehydrogenase, clinical stage, and histologic subtype. TGFβRII expression may be considered a new prognostic factor of diffuse large B cell lymphoma.

Topics: Adult; Aged; Biomarkers, Tumor; Female; Humans; Immunohistochemistry; Lymphoma, Large B-Cell, Diffuse; Male; Middle Aged; Prognosis; Protein Serine-Threonine Kinases; Receptor, Transforming Growth Factor-beta Type II; Receptors, Transforming Growth Factor beta; Signal Transduction; Survival Analysis; Transforming Growth Factor beta

Published data indicate that common genetic variants in immune/inflammatory response genes can affect the outcome of diffuse large B-cell lymphomas (DLBCL). This study investigated the association of interleukin (IL)-10 (-3575, -1082), tumor necrosis factor (TNF)-α -308 and transforming growth factor (TGF)-β Leu10Pro gene polymorphisms with clinical characteristics and outcome of DLBCL patients treated with rituximab-CHOP therapy.. Between January 2004 and December 2007, a total of 84 patients with newly diagnosed DLBCL entered into this study. Genotypes were determined with PCR-based methodology.. Patients presenting with B symptoms had IL-10 -3575 TA/AA genotypes more frequently than TT genotype [odds ratio (OR) 2.89, 95 % confidence interval (CI) 1.11-7.57; p = 0.03]. Carriers of TGF-β Pro10 allele more frequently had an advanced clinical stage III/IV (OR 4.65, 95 % CI 1.33-16.19; p = 0.016) and intermediate-high/high IPI score (OR 5.37, 95 % CI 1.45-20.0; p = 0.012). In rituximab-CHOP-treated patients (n = 64), the TNF-α -308 AG/AA carriers had shorter overall (p = 0.048) and event-free survival (p = 0.07) compared to GG carriers. In multivariate analysis of prognostic factors for survival, the TNF-α AG/AA genotypes were significantly associated with inferior survival of lymphoma patients (OR 0.23, 95 % CI 0.07-0.78; p = 0.018).. Our results indicate the association of IL-10 -3575 and TGF-β Leu10Pro gene variations with clinical characteristics. In patients treated with rituximab-CHOP therapy, the TNF-α -308 AG/AA genotypes showed a significantly less favorable survival than the GG genotype.

Topics: Aged; Antibodies, Monoclonal, Murine-Derived; Antineoplastic Combined Chemotherapy Protocols; Biomarkers, Pharmacological; Cyclophosphamide; Disease-Free Survival; Doxorubicin; Genetic Association Studies; Genetic Predisposition to Disease; Genotype; Humans; Interleukin-10; Kaplan-Meier Estimate; Lymphoma, Large B-Cell, Diffuse; Male; Middle Aged; Neoplasm Staging; Polymorphism, Single Nucleotide; Prednisone; Rituximab; Transforming Growth Factor beta; Treatment Outcome; Tumor Necrosis Factor-alpha; Vincristine

Neoplastic processes, tumor growth, and tumor cell proliferation and survival are often due to the altered activation of different signaling pathways. The increased activity of PI3K/AKT/mTOR signaling has been shown to be an important regulator of tumor growth in several solid tumors and in mantle cell lymphomas. The active form of mTOR kinase (mammalian target of rapamycin) is a key signaling molecule, and it exists in two different complexes, mTORC1 and mTORC2. In the present work, mTOR activity was investigated in different lymphoma types, in parallel with clinical data. We also examined in Hodgkin lymphomas (HL) the role of mTOR activity in survival mechanisms such as antiapoptotic protein expression and alterations in the microenvironment. We determined which lymphoma types display characteristic high mTOR activity in our TMA (tissue microarray) study. We observed that mTOR activity is increased in mitotic lymphoid cells compared to interphasic cells. The number of diffuse large B cell lymphoma (DLBCL) and HL cases was extended in a further set of TMA. We observed significantly higher mTOR activity in the non-centrum germinativum derived subtype of DLBCL than in the centrum germinativum derived subtype, which was a prognostic marker; 63% of mTOR active cases showed Rictor overexpression, indicating mTORC2 activity. High mTOR activity was also established in 92% of HL cases, which was linked to mTORC1. This finding was not a prognostic marker, however, it can be useful in targeted therapy. We observed the overexpression of the antiapoptotic protein BCL-xL and NFκB-p50 in the majority of mTOR active HLs. HLs showed high numbers of regulatory T cells in the microenvironment and high expression of galectin-1 in tumor cells and in the extracellular matrix, when compared to reactive lymph nodes. We confirmed that mTOR inhibition had significant antiproliferative and antiapoptotic effects in lymphoma cell lines and in lymphoma xenografts (HL, DLBCL, Burkitt lymphoma). We also showed that rapamycin was able to augment the effect of chemotherapeutic agents and TGF-β. Taken together, mTOR activity may be a potential therapeutic target in different lymphoma types. However, patient and inhibitor selection criteria must be carefully considered. The combination of mTOR inhibitors with other agents will probably offer the highest efficiency for achieving the best clinical response, and may also allow dose reduction in order to decrease late treatment toxicity in th. A neopláziás folyamatok kialakulása, a daganat növekedése, a daganatsejtek proliferációja és túlélése hátterében gyakran különbözõ jelutak magváltozott aktivitása áll. A PI3K/AKT/mTOR jelút fokozott aktivitása szolid daganatokban és köpenysejtes lymphomákban a daganatkialakulás és -növekedés fontos szabályozója. Az aktív mTOR (mammalian target of rapamycin) kináz két komplex (mTORC1, mTORC2) meghatározó eleme. Az mTOR-aktivitás szerepét vizsgáltuk különbözõ humán lymphomákban, összefüggéseket keresve a betegek klinikai adataival. Hodgkin-lymphomákban (HL) tanulmányoztuk, hogy a magas mTOR-aktivitás milyen, a daganat túlélésében fontos folyamatokban vesz részt (antiapoptotikus mechanizmusok és mikrokörnyezeti változások). Meghatároztuk azokat a humán lymphomatípusokat, amelyekre magas mTOR-aktivitás jellemzõ. Kimutattuk, hogy a mitotikus lymphoid sejtek mTOR-aktivitása magasabb, mint a nem osztódó sejteké. Nagyobb esetszámot tartalmazó TMA-blokkokon (tissue microarray) tovább vizsgáltuk a diffúz nagy B-sejtes lymphoma (DLBCL) és a HL eseteket. Szignifikáns összefüggést mutattunk ki DLBCL-s betegek altípusmegoszlása (csíraközpont-eredetû és nem csíraközpont-eredetû DLBCL-ek) és az mTOR-aktivitás között. DLBCL-ban a fokozott mTOR-aktivitás negatív prognosztikus markernek bizonyult. A HL-ek 92%-a magas mTOR-aktivitást mutatott (mTORC1-hez köthetõ), ami prognosztikus faktorként nem, viszont terápiás célpontként felhasználható. A HL-ek mikrokörnyezetének vizsgálata szerint a regulátor T-sejtek mennyisége a mikrokörnyezetben, valamint a galektin-1-expresszió a tumorsejtekben és az extracelluláris mátrixban emelkedett. A magas mTOR-aktivitás és a galektin-1-expresszió között kapcsolatot találtunk in vitro kísérleteinkben, ahol az mTOR gátlása transzlációs szinten csökkentette a galektin-1-expressziót. Az mTOR-gátlás jelentõségét – proliferációgátló és apoptotikus hatását – humán lymphoma xenograftokban (HL, DLBCL, Burkitt-lymphoma) bizonyítottuk. In vitro kombinációs kezelésekben a rapamycin apoptotikus hatást fokozó szerepét igazoltuk. Munkánkban meghatároztuk azokat a lymphomatípusokat, amelyekben az mTOR-gátlás célzott terápiaként alkalmazható lehet. Eredményeink alapján annak meghatározása, hogy melyik komplexhez köthetõ az mTOR-aktivitás, nagyon fontos a megfelelõ mTOR-gátló (klasszikus vagy kettõs gátlók) kiválasztásában. A jövõben a magas mTOR-aktivitást mutató lymphomákban várhatóan kombinációs kezelésben az mTOR-gátlók használata hozzájárulhatna a jobb t

Topics: Animals; Antineoplastic Agents; Apoptosis; Apoptosis Regulatory Proteins; bcl-X Protein; Burkitt Lymphoma; Cell Line; Drug Synergism; Galectin 1; Gene Expression Regulation, Neoplastic; Hodgkin Disease; Humans; Immunosuppressive Agents; Interphase; Lymphoma; Lymphoma, Large B-Cell, Diffuse; Mechanistic Target of Rapamycin Complex 1; Mechanistic Target of Rapamycin Complex 2; Mitosis; Multiprotein Complexes; NF-kappa B p50 Subunit; Signal Transduction; Sirolimus; T-Lymphocytes, Regulatory; Tissue Array Analysis; TOR Serine-Threonine Kinases; Transforming Growth Factor beta; Up-Regulation; Xenograft Model Antitumor Assays

This study was purposed to detect the expression levels of TRAF6, TAK1 and TGF-β mRNA in peripheral blood mononuclear cell (PBMNC) of patients with diffuse large B cell lymphoma (DLBCL) before and after chemotherapy, and to explore the effect of chemotherapy on the activity of TRAF6/TAK1 signal pathway. The expression levels of TRAF-6, TAK1 and TGF-β mRNA in PBMNC of 38 patients with DLBCL were detected by using the quantitative real time PCR before treatment or after two cycles of chemotherapy, 12 healthy people were served as the control. The results showed that the expression levels of TRAF-6, TAK1 and TGF-β mRNA in PBMNC of DLBCL patients' were higher than those in healthy people. Before treatment, the expression levels of TRAF-6 and TAK1 mRNA had no significant difference as compared with healthy people (P > 0.05); after chemotherapy, the expression levels of these two genes significantly increased, and the differences both had statistically significant as compared with healthy people (P < 0.05); meanwhile the increased expression levels of these two genes after chemotherapy had statistically significant difference as compared with levels before treatment (P < 0.05) , and those expression levels were positively correlated. While the expression level of TGF-β mRNA decreased after chemotherapy as compared with level before treatment, and the differences had statistically significantse(P < 0.05). It is concluded that the activity of TRF6/TAK1 signal pathways in PBMNC of DLBCL patients' significantly increases after chemotherapy, while the expression level of TGF-β mRNA after chemotherapy is abviously lower than level before treatment.

Topics: Gene Expression Regulation, Leukemic; Humans; Leukocytes, Mononuclear; Lymphoma, Large B-Cell, Diffuse; MAP Kinase Kinase Kinases; RNA, Messenger; Signal Transduction; TNF Receptor-Associated Factor 6; Transforming Growth Factor beta

The mechanisms by which microRNA dysfunction contributes to the pathogenesis of diffuse large B cell lymphoma (DLBCL) are not well established. The identification of the genes and pathways directly targeted by these small regulatory RNAs is a critical step to advance this field. Using unbiased genome-wide approaches in DLBCL, we discovered that the oncogenic microRNA-155 (miR-155) directly targets the bone morphogenetic protein (BMP)-responsive transcriptional factor SMAD5. Surprisingly, we found that in DLBCL a noncanonical signaling module linking TGF-beta1 signals to SMAD5 is also active. In agreement with these data, miR-155 overexpression rendered DLBCLs resistant to the growth-inhibitory effects of both TGF-beta1 and BMPs, via defective induction of p21 and impaired cell cycle arrest. In confirmatory experiments, RNAi-based SMAD5 knockdown recapitulated in vitro and in vivo the effects miR-155 overexpression. Furthermore, in primary DLBCLs, miR-155 overexpression inhibited SMAD5 expression and disrupted its activity, as defined by individual and global analyses of its transcriptional targets. Together, our data helped explain miR-155 function, highlighted a hitherto unappreciated role of SMAD5 in lymphoma biology, and defined a unique mechanism used by cancer cells to escape TGF-beta's growth-inhibitory effects.

Topics: Bone Morphogenetic Proteins; Cell Line, Tumor; Cyclin-Dependent Kinase Inhibitor p21; Humans; Lymphoma, Large B-Cell, Diffuse; MicroRNAs; Reverse Transcriptase Polymerase Chain Reaction; RNA Interference; Smad5 Protein; Statistics, Nonparametric; Transforming Growth Factor beta

Pulmonary fibrosis is a severe complication associated with bis-chloronitrosourea (BCNU) therapy. However, the pathogenetic mechanism has never been well investigated. We report here a 26-year-old female with diffuse large B-cell lymphoma who died of severe pulmonary fibrosis 81 days after the administration of high-dose BCNU (600 mg/m2). Thoracoscopic wedge resection of left upper lung performed 10 days before patient's death showed severe pulmonary fibrosis with prominent hyperplasia of alveolar macrophages and type II pneumocytes. We further used immunohistochemistry (IHC) to examine the relative role of platelet-derived growth factor-B (PDGF-B), insulin-like growth factor I (IGF-I), transforming growth factor-beta1 (TGF-beta1) and cyclooxygenase-2 (COX-2) in the pathogenesis of BCNU-related pulmonary fibrosis. Strong expressions of PDGF-B and IGF-1 on alveolar macrophages and type II pneumocytes were clearly demonstrated, but in contrast, the expressions of TGF-beta1 and COX-2 were almost undetectable. In conclusion, pulmonary fibrosis can develop early and progress rapidly after the administration of high-dose BCNU. The markedly increased expression of fibrogenic factors PDGF-B and IGF-1 on hyperplastic alveolar macrophages and hyperplastic type II pneumocytes may play an important role in the fibrogenesis of this disease. These novel findings may offer specific therapeutic targets in the treatment of BCNU-associated pulmonary fibrosis.

Topics: Adult; Antineoplastic Agents, Alkylating; Carmustine; Cyclooxygenase 2; Fatal Outcome; Female; Humans; Insulin-Like Growth Factor I; Isoenzymes; Lung; Lymphoma, B-Cell; Lymphoma, Large B-Cell, Diffuse; Membrane Proteins; Prostaglandin-Endoperoxide Synthases; Proto-Oncogene Proteins c-sis; Pulmonary Fibrosis; Transforming Growth Factor beta; Transforming Growth Factor beta1

With increasing awareness of the importance of renal cortical interstitial fibrosis, interest has focused on the mechanisms that stimulate generation of profibrotic factors including transforming growth factor (TGF)-beta1, by resident cells, such as proximal tubular epithelial cells (PTCs). Infiltration of monocytes, has been implicated in the pathogenesis of a wide variety of renal diseases, however, how interaction between monocytes and PTCs may affect the generation of TGF-beta1 by the resident cell is unknown. We demonstrate that monocytes stimulate TGF-beta1 transcription and protein synthesis by PTCs. This was dependent on direct cell contact and TGF-beta1 transcriptional activation that was dependent on ICAM-1 binding of unstimulated monocytes. This was mimicked by antibody cross-linking of PTC surface ICAM-1. We have previously identified hyaluronan (HA)-based structures on the surface of PTCs, both primary cultures and the HK-2 cell line. Removal of cell-surface HA increased ICAM-1-dependent monocyte binding and stimulation of TGF-beta1 synthesis. Furthermore, we demonstrate that binding of monocytes to HA-based structures on the cell surface of HK-2 cells interferes with this response. In summary, we have demonstrated that HA-based pericellular structures down-regulate proinflammatory and profibrotic responses by modulation of monocyte-driven ICAM-1-dependent cell activation and TGF-beta1 generation.

Topics: Cell Communication; Epithelial Cells; Humans; Hyaluronic Acid; Intercellular Adhesion Molecule-1; Kidney Tubules, Proximal; Luciferases; Lymphoma, Large B-Cell, Diffuse; Monocytes; Promoter Regions, Genetic; Protein Binding; Transcription, Genetic; Transforming Growth Factor beta; Transforming Growth Factor beta1; U937 Cells

We have previously reported an overexpression of Smad1 in follicular lymphoma (FL) cells, which are characterized by the t(14;18) bcl2/IgH translocation. Smad1 is commonly involved in bone morphogenetic protein but not in tumor-transforming growth factor beta (TGFbeta) signaling pathways. This study focuses on Smad1 signaling pathway in non-Hodgkin lymphoma cells including follicular or large-cell lymphoma cells. Our results support the notion that phosphorylation of Smad1 is mediated by TGFbeta present in the microenvironment and occurs in FL in vivo. Using an in vitro coculture system mimicking interactions between stroma cells and FL cells, we found that both the cell partners release TGFbeta at a sufficient concentration to activate Smad pathways in the malignant cells. This Smad1 activation involves TGFbetaRII but not ALK-1 receptors, and does not compete with the Smad2 pathway. Moreover, proliferation assays performed on lymphoma cells expressing wild-type or mutated Smad1, or in which endogenous Smad1 level was decreased by gene silencing, strongly supported that overexpression and activation of Smad1 modifies the biological response of lymphoma B cells to TGFbeta family members. This work opens new insights into aberrant Smad pathways and their pathophysiological role in FL and in other non-Hodgkin lymphomas.

Topics: Activin Receptors, Type I; Activin Receptors, Type II; B-Lymphocytes; Cell Proliferation; DNA-Binding Proteins; Gene Silencing; Humans; Lymphoma, B-Cell; Lymphoma, Follicular; Lymphoma, Large B-Cell, Diffuse; Mutation; Palatine Tonsil; Phosphorylation; Protein Serine-Threonine Kinases; Receptor, Transforming Growth Factor-beta Type II; Receptors, Transforming Growth Factor beta; Signal Transduction; Smad Proteins; Smad1 Protein; Smad2 Protein; Stromal Cells; Trans-Activators; Transforming Growth Factor beta; Tumor Cells, Cultured

The retinoblastoma tumor-suppressor gene, RB, has been implicated in tumor suppression, in regulation of the cell cycle, and in mediating cell differentiation. RB is necessary for hematopoiesis in mice, and aberrant RB-expression is associated with the progress and prognosis of leukemia. We have used antisense oligonucleotides, established clones stably expressing an antisense RB construct, and also established clones over expressing the retinoblastoma protein (pRb) to study the role of RB expression in monocytic differentiation induced by all-trans retinoic acid (ATRA) or 1-alpha-25-dihyroxycholecalciferol (Vit D3) in the monoblastic cell line U-937 and erythroid differentiation induced by transforming growth factor beta1 (TGFbeta1) and hemin in the erythroleukemic cell line K562. A reduction in pRb production in antisense RB-transfected U-937 clones was shown. Antisense oligonucleotides as well as expression of the antisense RB construct suppressed differentiation responses to ATRA or Vit D3, as judged by the capability to reduce nitro blue tetrazolium, by the appearance of monocyte-related cell surface antigens and by morphologic criteria. K562 cells showed decreased differentiation response to TGFbeta1, but not to hemin, when incubated with antisense oligonucleotides. U-937 antisense RB-transfected cells were also suppressed in their ability to upregulate levels of hypophosphorylated pRb when induced to differentiate. Although U-937 cells incubated with antisense oligonucleotides and clones expressing the antisense RB construct were hampered in their ability to differentiate on incubation with ATRA or Vit D3, the induced G0/G1-accumulation was similar to differentiating control cells treated with ATRA or Vit D3. Intriguingly, U-937 clones overexpressing RB were also inhibited in their differentiation response to ATRA or Vit D3 but not inhibited in their ability to respond with G0/G1 accumulation when induced with these substances. The results indicate that pRb plays a role in induced differentiation of U-937 cells as well as K562 cells involving mechanisms that, at least partially, are distinct from those inducing G1 accumulation.

Topics: Animals; Antigens, Differentiation; Antigens, Neoplasm; Calcitriol; Cell Differentiation; G1 Phase; Gene Expression Regulation, Leukemic; Genes, Retinoblastoma; Leukemia; Leukemia, Myelogenous, Chronic, BCR-ABL Positive; Lymphoma, Large B-Cell, Diffuse; Mice; Monocytes; Neoplasm Proteins; Oligonucleotides, Antisense; Retinoblastoma Protein; Transfection; Transforming Growth Factor beta; Tretinoin; Tumor Cells, Cultured

The leukocyte integrin Mac-1 (CD11b/CD18) and the urokinase receptor (uPAR, CD87) mediate complementary functions in myelomonocytic cells. Both receptors promote degradation of fibrin(ogen) and also confer adhesive properties on cells because Mac-1 and uPAR bind fibrin and vitronectin, respectively. Staining of lung biopsy specimens from patients with acute lung injury indicated that fibrin and vitronectin colocalize at exudative sites in which macrophages bearing these receptors accumulate. Because of the parallel roles and physical proximity of Mac-1 and uPAR, the capacity of these receptors to functionally interact was explored. Induction of Mac-1 and uPAR expression on monocytic cell lines by transforming growth factor- beta 1 and 1.25-(OH)2 vitamin D3 conferred urokinase and uPAR-dependent adhesion to vitronectin, which was further promoted by engagement of Mac-1. Vitronectin attachment promoted subsequent Mac-1-mediated fibrinogen degradation threefold to fourfold. In contrast, enhancement of uPAR occupancy by exogenous urokinase or receptor binding fragments thereof inhibited Mac-1 function. Addition of urokinase progressively inhibited Mac-1-mediated fibrinogen binding and degradation (maximal inhibition, 91% +/- 14% and 72% +/- 15%, respectively). Saturation of uPAR with urokinase also inhibited binding of the procoagulant Mac-1 ligand, Factor X. These inhibitory effects of uPAR were reproduced in fresh monocytes, cultured monocytic cells, and in Chinese hamster ovary (CHO) cells transfected with both human Mac-1 and human uPAR. These data show that the procoagulant and fibrinolytic potential of monocytic cells is co-ordinately regulated by ligand binding to both Mac-1 and uPAR and identify uPAR as a regulator of integrin function. Vitronectin-enhanced fibrin(ogen) turnover by Mac-1 may operate as a salvage pathway in the setting of urokinase and plasmin inhibitors to promote clearance of the provisional matrix and subsequent healing.

Topics: Animals; Blood Coagulation; Calcitriol; CD18 Antigens; Cell Adhesion; CHO Cells; Cricetinae; Factor X; Fibrin; Fibrinogen; Fibrinolysis; Humans; Leukemia, Monocytic, Acute; Lung; Lung Injury; Lymphoma, Large B-Cell, Diffuse; Macromolecular Substances; Macrophage-1 Antigen; Macrophages; Monocytes; Neoplasm Proteins; Receptors, Cell Surface; Receptors, Urokinase Plasminogen Activator; Recombinant Proteins; Transfection; Transforming Growth Factor beta; Tumor Cells, Cultured; Urokinase-Type Plasminogen Activator; Vitronectin

We found that production and release of two functionally antagonistic cytokines, TGF-beta and TNF-alpha, were regulated differently in the mast cell, T cell, and macrophage cell lines RBL-2H3, MLA-144, and U-937, respectively. TGF-beta was produced and released constitutively in all three cell lines. When, however, the cell lines were stimulated with Ag, LPS, or calcium ionophore plus PMA, acceleration of release and some additional production of TGF-beta were apparent. In contrast, TNF-alpha was produced and released only when these lines were stimulated. Although neither the glucocorticoid, dexamethasone, nor the protein kinase C inhibitor, Ro31-7549, suppressed constitutive production or release of TGF-beta, these agents inhibited TNF-alpha production and the inducible component of TGF-beta production noted above. The release of these cytokines, whether constitutive or inducible, was dependent on Golgi-processing as indicated by inhibition with brefeldin A. Therefore, although both types of cytokines were processed by Golgi, only TNF-alpha and the inducible component of TGF-beta production were protein kinase C or steroid-regulated processes. These findings suggested that constitutive and inducible pathways exist for production and release of cytokines and that the inducible pathways can be selectively suppressed by pharmacologic agents.

Topics: Animals; Brefeldin A; Cyclopentanes; Dexamethasone; Enzyme Inhibitors; Gene Expression Regulation; Golgi Apparatus; Humans; Indoles; Leukemia, Basophilic, Acute; Lymphocyte Activation; Lymphoma, Large B-Cell, Diffuse; Macrophage Activation; Macrophages; Maleimides; Mast Cells; Protein Kinase C; Rats; Signal Transduction; T-Lymphocytes; Transforming Growth Factor beta; Tumor Cells, Cultured; Tumor Necrosis Factor-alpha

Osteolysis has become a major cause of aseptic loosening in total joint arthroplasty (TJA). Titanium, cobalt and chromium are commonly used in orthopaedic implants (e.g. joint prostheses). The release of bone-associated cytokines has been associated with the development of osteolysis in patients with prostheses. We evaluated the effects of these metals on the release of bone-associated cytokines (IL-1 beta, IL-6, TNF-alpha and TGF-beta 1) by human blood monocytes/macrophages and monocyte-like U937 cells upon lipopolysaccharide (LPS) stimulation, the cell proliferation, and their cytotoxic effects on these cells in vitro. We found that the release of IL-1 beta was enhanced by titanium, chromium and cobalt, the release of TNF-alpha was enhanced by titanium and chromium, and the release of IL-6 was enhanced by titanium. All three metal ions inhibited the release of TGF-beta 1. We also found that titanium and chromium, but not cobalt, enhanced blood monocyte/macrophage proliferation in response to LPS while only titanium enhanced U937 cell proliferation in response to LPS. The metals in concentrations ranging from 0.01 to 100 ngml-1 did not stimulate the cells to secrete detectable cytokines in the absence of LPS. Furthermore, a 4-h pre-exposure of blood monocytes/macrophages or U937 cells to the metals did not alter cytokine release when the metals were removed from the media prior to the addition of LPS. Similarly, a 4-h pre-exposure of blood monocytes/macrophages or U937 cells to LPS did not alter cytokine release when LPS was removed from the media prior to the addition of the metals. The metals did not reduce cell viability and induce cell injury after 72h incubation with the cells. The data suggest that the three metals at clinically relevant concentrations modulated cytokine expression, whereas they did not induce any cytotoxic effects. A metal-induced enhancement of bone-resorbing cytokine release with a concomitant inhibition of bone-forming cytokine release may be an important factor in the development of osteolysis, which can severely compromise the outcome of TJA.

Topics: Biocompatible Materials; Cell Division; Cells, Cultured; Chromium; Cobalt; Corrosion; Cytokines; Humans; Interleukin-1; Interleukin-6; Joint Prosthesis; Lymphoma, Large B-Cell, Diffuse; Macrophages; Monocytes; Osteolysis; Prosthesis Failure; Titanium; Transforming Growth Factor beta; Tumor Cells, Cultured; Tumor Necrosis Factor-alpha

Patients with HIV infection often develop glomerular lesions (focal segmental glomerular sclerosis). Because mesangial expansion (enhanced mesangial cell (MC) growth and matrix accumulation) has been demonstrated to precede the development of focal segmental glomerulosclerosis, we studied the effect of the interaction between HIV-1 proteins such as gp160 envelope protein and macrophages on mesangial cell proliferation and matrix synthesis. We determined the effect of control media, serum-free macrophage supernatant (MSP), and serum-free HIV-1 gp 160 protein-treated MSP (gp 160-MSP) on the proliferation of MC and synthesis of collagen type IV (a component of mesangial matrix). MSP (20%) enhanced (P < 0.01) MC proliferation (control, 7.58 +/- 0.29 versus MSP, 9.06 +/- 0.25 x 10(4) cells/ml), whereas gp 160-MSP (20%) inhibited (P < 0.001) MC proliferation (gp160-MSP, 5.58 +/- 0.14 x 10(4) cells/ml). gp160-MSP modulated MC proliferation in a dose-dependent manner; it enhanced cell proliferation at a lower concentration but inhibited cell proliferation at a higher concentration. Anti-TGF-beta antibody attenuated the effect of gp160-MSP on MC proliferation at lower as well as higher concentrations. Bromodeoxyuridine incorporation studies also showed the modulation of MC proliferation by gp160-MSP. Interaction of other HIV proteins such as HIV-1 Gag4 and HIV-1 Tat with macrophages did not affect MC proliferation when compared with MSP alone. gp160-MSP also enhanced (P < 0.001) synthesis of type IV collagen by MC (control, 467.8 +/- 9.0; MSP, 501.0 +/- 25.0; gp160-MSP, 775.5 +/- 39.0 ng/mg protein). The effect of gp160-MSP on collagen synthesis by MC was dose-dependent. Anti-TGF-beta antibody attenuated the gp160-MSP-induced mesangial cell collagen synthesis. The present study provides a basis for speculation that macrophage-gp160 interaction products have the potential to cause expansion of the mesangium.

Topics: Animals; Antibodies, Monoclonal; Cell Division; Cell Line; Culture Media, Conditioned; Extracellular Matrix; Gene Products, env; Gene Products, gag; Gene Products, tat; Glomerular Mesangium; HIV Envelope Protein gp160; Humans; Leukocytes, Mononuclear; Lymphoma, Large B-Cell, Diffuse; Macrophages; Mice; Protein Precursors; Transforming Growth Factor beta; Tumor Cells, Cultured

We wished to examine the role of transforming growth factor-beta (TGF-beta) in the regulation of human lymphoma cell growth. The RL cell line is an immunoglobulin M (IgM)+, IgD+ B lymphoma cell line, which does not constitutively express receptors for TGF-beta, and thus has lost the ability to respond to the inhibitory effects of TGF-beta. We demonstrate here that anti-Ig antibodies can efficiently upregulate the expression of TGF-beta receptors and promote sensitivity to growth inhibition by TGF-beta. Furthermore, because TGF-beta has been shown to function in late G1 of the cell cycle, we examined the ability of TGF-beta to modulate two tumor suppressor proteins known to be critical regulators of the G1/S transition, Rb and p53. Rb is a 105- to 110-kD phosphoprotein, which has been shown to maintain its growth suppressive function when it is found in the hypophosphorylated state. Wild-type p53 is a 53-kD phosphoprotein that appears to be important in preventing cell-cycle progression and promoting apoptosis in cells with DNA damage, whereas mutant p53 can overcome those functions. We show here that TGF-beta treatment of phorbol myristate acetate (PMA) or anti-Ig-activated RL cells results in growth inhibition through a dual effect on Rb and mutant p53. After TGF-beta treatment, we observe a predominance of Rb in the hypophosphorylated, growth suppressive form. In addition, we show a decrease in levels of mRNA and protein for mutant p53. We also show that, although these changes are sufficient to halt progression through the cell cycle, the cells do not appear to undergo extensive programmed cell death following 72 hours of TGF-beta treatment. Thus, although these lymphoma cells maintain the capacity to be negatively growth regulated by TGF-beta, the ability of TGF-beta to induce apoptosis must be independently controlled.

Topics: Animals; Antibodies, Anti-Idiotypic; Apoptosis; Cell Division; Drug Resistance; G1 Phase; Gene Expression Regulation, Neoplastic; Humans; Immunoglobulin M; Lymphoma, Large B-Cell, Diffuse; Neoplasm Proteins; Rabbits; Receptors, Antigen, B-Cell; Receptors, Transforming Growth Factor beta; Retinoblastoma Protein; Tetradecanoylphorbol Acetate; Transforming Growth Factor beta; Tumor Cells, Cultured; Tumor Suppressor Protein p53; Up-Regulation

A patient with chronic lymphocytic leukemia developed a large cell lymphoma apparently derived from the same neoplastic B-cell clone (Richter's syndrome). At the same time, mitogen-stimulated proliferation of the patient's circulating leukemic B-cells was no longer inhibited by the regulatory cytokine transforming growth factor-beta (TGF-beta), suggesting that such loss of inhibition might be contributing to the clinical and biological progression of the disease.

Topics: Aged; B-Lymphocytes; Cell Division; Depression, Chemical; Humans; Leukemia, Lymphocytic, Chronic, B-Cell; Lymphoma, B-Cell; Lymphoma, Large B-Cell, Diffuse; Male; Syndrome; Transforming Growth Factor beta; Tumor Cells, Cultured