Clinical success of the proteasome inhibitor established bortezomib as one of the most effective drugs in treatment of multiple myeloma (MM). While survival benefit of bortezomib generated new treatment strategies, the primary and secondary resistance of MM cells to bortezomib remains a clinical concern. This study aimed to highlight the role of p53-induced RING-H2 (Pirh2) in the acquisition of bortezomib resistance in MM and to clarify the function and mechanism of action of Pirh2 in MM cell growth and resistance, thereby providing the basis for new therapeutic targets for MM. The proteasome inhibitor bortezomib has been established as one of the most effective drugs for treating MM. We demonstrated that bortezomib resistance in MM cells resulted from a reduction in Pirh2 protein levels. Pirh2 overexpression overcame bortezomib resistance and restored the sensitivity of myeloma cells to bortezomib, while a reduction in Pirh2 levels was correlated with bortezomib resistance. The levels of nuclear factor-kappaB (NF-κB) p65, pp65, pIKBa, and IKKa were higher in bortezomib-resistant cells than those in parental cells. Pirh2 overexpression reduced the levels of pIKBa and IKKa, while the knockdown of Pirh2 via short hairpin RNAs increased the expression of NF-κB p65, pIKBa, and IKKa. Therefore, Pirh2 suppressed the canonical NF-κB signaling pathway by inhibiting the phosphorylation and subsequent degradation of IKBa to overcome acquired bortezomib resistance in MM cells.
Antineoplastic Agents ; pharmacology ; therapeutic use ; Apoptosis ; drug effects ; Bortezomib ; pharmacology ; therapeutic use ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Dose-Response Relationship, Drug ; Drug Resistance, Neoplasm ; drug effects ; Drug Screening Assays, Antitumor ; Humans ; Multiple Myeloma ; drug therapy ; metabolism ; pathology ; NF-kappa B ; metabolism ; Signal Transduction ; drug effects ; Structure-Activity Relationship ; Ubiquitin-Protein Ligases ; genetics ; metabolism

No abstract available.
Aged ; Antineoplastic Agents/therapeutic use ; Bone Marrow/pathology ; Fusion Proteins, bcr-abl/genetics/metabolism ; Humans ; Imatinib Mesylate/therapeutic use ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive/complications/*diagnosis/drug therapy ; Leukocyte Count ; Male ; Multiple Myeloma/complications/*diagnosis/drug therapy ; Platelet Count ; Polymerase Chain Reaction ; Thrombocytosis/etiology

ADP-ribosyl Cyclase 1 ; metabolism ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Biomarkers, Tumor ; metabolism ; Biopsy ; Bone Neoplasms ; drug therapy ; metabolism ; pathology ; CD56 Antigen ; metabolism ; Humans ; Ilium ; Interferon Regulatory Factors ; Male ; Middle Aged ; Multiple Myeloma ; drug therapy ; metabolism ; pathology ; Syndecan-1 ; metabolism

The purpose of this study was to investigate the effect of bisphosphonate combined with chemotherapy on bone mineral density (BMD) of patients with multiple myeloma (MM) and analyse its value of BMD detection in clinic of these patients. 53 MM cases were enrolled in this study, including 33 newly diagnosed, 10 refractory/relapsed and 10 stable cases. They were divided randomly into two groups, 33 MM cases were treated with bisphosphonates combined with chemotherapy and 20 MM cases were treated with chemotherapy alone. The chemotherapy schedules for all patients were same. BMD was tested using the dual energy X-ray absorptiometry at 2 time points, i.e. pretreatment (basal level) and 12 months after treatment with chemotherapy and bisphosphonates. Comparisons was performed with t tests using SPSS 11.0 software. The results indicated that there was minor difference between 2 groups for BMD scores of whole body and lumbar vertebra (L1-L4), but no difference for scores of the near-end of left femur. After treatment for 12 months, all BMD scores (whole body, lumbar vertebra and the near-end of left femur) increased significantly in the bisphosphonate combined with chemotherapy group (P < 0.05). In contrast, only minor changes were seen in chemotherapy alone group. It is concluded that the bisphosphonate combined with chemotherapy has displayed promotive effect on BMD of MM patients, the detection of BMD is sensitive and special method for monitoring therapeutic effect of bisphosphonate in MM patients, thus has useful value in clinic of these patients.
Adult ; Aged ; Antineoplastic Agents ; therapeutic use ; Bone Density ; drug effects ; Diphosphonates ; administration & dosage ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; drug therapy ; metabolism ; pathology

We report a rare case of multiple myeloma with biclonal gammopathy (IgG kappa and IgA lambda type) in a 58-year-old man with prostate cancer who presented with lower back pain. Through computed tomography (CT) imaging, an osteolytic lesion at the L3 vertebra and an enhancing lesion of the prostate gland with multiple lymphadenopathies were found. In the whole body positron emission tomography-computed tomography (PET-CT), an additional osteoblastic bone lesion was found in the left ischial bone. A prostate biopsy was performed, and adenocarcinoma was confirmed. Decompression surgery of the L3 vertebra was conducted, and the pathologic result indicated that the lesion was a plasma cell neoplasm. Immunofixation electrophoresis showed the presence of biclonal gammopathy (IgG kappa and IgA lambda). Bone marrow plasma cells (CD138 positive cells) comprised 7.2% of nucleated cells and showed kappa positivity. We started radiation therapy for the L3 vertebra lesion, with a total dose of 3,940 cGy, and androgen deprivation therapy as treatment for the prostate cancer.
Adenocarcinoma/complications/*diagnosis/radiotherapy ; Antineoplastic Agents/therapeutic use ; Bone Marrow Cells/metabolism/pathology ; Combined Modality Therapy ; Humans ; Immunoelectrophoresis ; Immunoglobulin kappa-Chains/blood ; Immunoglobulin lambda-Chains/blood ; Male ; Middle Aged ; Multiple Myeloma/complications/*diagnosis/drug therapy ; Neoplasm Staging ; Positron-Emission Tomography ; Prostatic Neoplasms/complications/*diagnosis/radiotherapy ; Spine/pathology ; Syndecan-1/metabolism ; Tomography, X-Ray Computed

Involvement of the central nervous system is very uncommon in multiple myeloma, observed in approximately 1% of the multiple myeloma patients. We report a case of central nervous system myelomatosis with complex chromosome aberrations in a 62-yr-old female patient, who had previously been diagnosed as multiple myeloma. Fluorescent in situ hybridization revealed 13q deletion, p53 gene deletion and IGH/FGFR3 rearrangement and chromosomal study showed complex chromosome aberrations. After four cycles of chemotherapy, the patient was admitted to the hematology department with severe headache. Plasma cells were found in the cerebrospinal fluid (CSF), and CSF immunoelectrophoresis revealed abnormal precipitin arcs against anti-IgG and anti-lambda antisera. She was given systemic chemotherapy and eight courses of intrathecal chemotherapy, which cleared plasma cells in the CSF. Two months later, she was given autologous stem cell transplantation. Three months after stem cell transplantation, central nervous system myelomatosis progressed to plasma cell leukemia and two months later,the patient expired.
Antineoplastic Agents/therapeutic use ; Central Nervous System Neoplasms/*diagnosis/drug therapy/genetics ; Cerebrospinal Fluid/cytology ; *Chromosome Deletion ; Combined Modality Therapy ; Disease Progression ; Female ; Gene Deletion ; Humans ; Immunoelectrophoresis ; In Situ Hybridization, Fluorescence ; Leukemia, Plasma Cell/diagnosis ; Middle Aged ; Multiple Myeloma/*diagnosis/drug therapy/genetics ; Plasma Cells/pathology ; Precipitins/metabolism ; Receptor, Fibroblast Growth Factor, Type 3/genetics ; Stem Cell Transplantation ; *Translocation, Genetic ; Transplantation, Autologous ; Tumor Suppressor Protein p53/genetics

OBJECTIVETo investigate the reverse effect of mutidrug resistance of curcumin combined with melphalan on the mutidrug-resistant human multiple myeloma cell line MOLP-2/R and the relation with FA/BRCA pathway.

METHODSThe inhibitory effects of the drugs on the growth of MOLP-2/R cells were determined by MTT assay. Cell cycle analysis, intracellular drug concentration and apoptosis were assayed by flow cytometry. The expression of FANCD2 monoubiquitination was determined by Western blot analysis.

RESULTSCo-administration of curcumin and melphalan had an synergistic inhibitory effects on the proliferation, IC50 of melphalan with 10 micromol/L curcumin reduced from 45.5 micromol/L to 19 micromol/L in MOLP-2/R cells. The apoptosis percentage of MOLP-2/R cells was significantly increased from (23.3 +/- 0.6)% to (52.6% +/- 0.8)% by the treatment of melphalan 20 micromol/L plus curcumin 10 micromol/L with the increased percentage of cells in the G2/M phase (from 9.1% to 18.5%) and enhanced intracellular drug concentration of MOLP-2/ R cells (from 15.2 +/- 0.3 to 21.4 +/- 0.8 ). The effects were accompanied with inhibition of FA/BRCA pathway by down regulation of FANCD2 protein monoubiquitination.

CONCLUSIONCurcumin combined with melphalan results in synergistic effects and reverses multiple drug resistance of MOLP-2/R cells effectively. The inhibition of FA/BRCA pathway may be the mechanism.

Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Curcumin ; pharmacology ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; drug effects ; Fanconi Anemia Complementation Group D2 Protein ; metabolism ; Humans ; Multiple Myeloma ; drug therapy ; metabolism ; pathology

OBJECTIVETo investigate the expression of B7 co-stimulatory molecules in human multiple myeloma (MM) and the immunoregulatory effects of thalidomide on B7.1 co-stimulator.

METHODSThe immunoregulatory effects of thalidomide on the expression of B7-1 in human MM cell line was examined by detecting the changes in the expression of B7 co-stimulator on the cells using flow cytometry following the drug treatment.

RESULTSThe expression of B7.1 co-stimulator was lowly expressed in human MM cell line and MM patients, with a positivity rate of 0.8 and (2.19-/+2.13) for B7.1 and a rate of 26.4 and (30.28-/+28.11) for B7.2, respectively. Compared with the control group, the thalidomide-treated cells showed significantly increased percentage of CD-80 positive cells in a dose-dependent manner (but not at 0.1 microg/ml) (P<0.01), with the highest percentage reaching (17.7-/+1.53)% at thalidomide concentration of 5 microg/ml.

CONCLUSIONMM cells express low or undetectable levels of B7.1. Thalidomide can up-regulate the expression of B7-1 molecules on myeloma cells, which is probably one of the therapeutic mechanisms of thalidomide.

Adult ; Aged ; Aged, 80 and over ; B7-1 Antigen ; drug effects ; metabolism ; Cell Line, Tumor ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; drug therapy ; immunology ; pathology ; Thalidomide ; therapeutic use ; Up-Regulation ; drug effects

OBJECTIVETo determine fatty acid synthase (FAS) expression in human multiple myeloma and verify its potential as a therapeutic target in multiple myeloma.

METHODSFAS expression was determined by immunohistochemistry, reverse-transcription polymerase chain reaction (RT-PCR) and immunoblot analysis in bone marrow samples obtained from 27 patients with multiple myeloma (MM patients) and peripheral blood mononuclear cells (PBMCs) obtained from 12 healthy donors. In parallel, additional analyses were performed on 2 human multiple myeloma cell lines, U266 and RPMI8226. U266 cells were treated with cerulenin at various concentrations (5 to 320 microg/ml) for 24 h, and metabolic activity was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. Apoptosis was evaluated by dual Annexin V/PI (propidium iodide) labeling and flow cytometry (FCM) in U266 cells treated with 20 (g/ml cerulenin for 12 h or 24 h.

RESULTSBy immunohistochemistry, we found that 19 of 27 bone marrow samples obtained from MM patients expressed significantly high levels of FAS. Similarly, by RT-PCR, 22 of 27 bone marrow samples obtained from MM patients, U266 and RPMI8226 showed FAS expression, whereas PBMC samples from 12 healthy donors did not express detectable level of FAS. FAS protein expression was confirmed by immunoblot analysis in 16 of 27 bone marrow samples obtained from MM patients, U266 and RPMI8226 cell lines, and no FAS protein expression was detected in PBMC samples from 12 healthy donors. U266 cells were highly sensitive to cerulenin treatment, with a dosage-related effect on metabolic activity, as a measure for cell proliferation. U266 cells treated with 20 microg/ml cerulenin for 12 and 24 h also showed early sign of apoptosis with 56.9% and 69.3% Annexin V(+)/PI(-) cells, and late apoptotic and necrotic cells with 3.2% and 17.6% Annexin V(+)/PI(+) cells.

CONCLUSIONIncreased FAS expression existed in multiple myeloma samples and human myeloma cell lines. Cerulenin greatly inhibited metabolic activity/cell proliferation of U266 cells and induced apoptosis, suggesting that FAS is an effective target for pharmacological therapy in human multiple myeloma.

Adult ; Aged ; Apoptosis ; drug effects ; Base Sequence ; Blotting, Western ; Case-Control Studies ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cerulenin ; pharmacology ; DNA Primers ; genetics ; Enzyme Inhibitors ; pharmacology ; Fatty Acid Synthase, Type I ; antagonists & inhibitors ; genetics ; metabolism ; Fatty Acid Synthesis Inhibitors ; pharmacology ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Multiple Myeloma ; drug therapy ; enzymology ; genetics ; pathology ; RNA, Messenger ; genetics ; metabolism ; RNA, Neoplasm ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

This study was aimed to further explore whether brain derived neurotrophic factor (BDNF) pathway is a potential therapeutic target in multiple myeloma (MM) and whether anti-BDNF monoclonal antibody can prevent the development of this disease. The in vivo antitumor effect of anti-BDNF monoclonal antibody (McAb) on a human myeloma xenograft animal model was evaluated. The model of xenograft tumors was established in the nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice by subcutaneous injection of human myeloma cell line RPMI8226. The antibodies were injected intraperitoneally at a dose of 20 microg/mouse at day 1, 2, 3 after inoculation or at a dose of 100 microg/mouse once a week after tumors were detected. The microvascular densities in tumors were analyzed by immunohistochemistry study. The effect of anti-BDNF McAb on the proliferation of RPMI8226 cells in vitro and on endothelial cells network formation in the co-culture system were determined by using a (3)H-thymidine incorporation assay and a Matrigel network formation assay, respectively. The results showed that multiple injections of anti-BDNF McAb reduced the tumor size, decreased the microvascular density and significantly prolonged tumor-free time and survival time. Moreover, the proliferation of RPMI8226 cells was inhibited in vitro by anti-BDNF McAb, but not by the control IgG. Anti-BDNF McAb also inhibited RPMI8226-induced network formation in endothelial cells in vitro. It is concluded that anti-BDNF monoclonal antibody can inhibit cell growth and angiogenesis in subcutaneous plasmacytoma.
Animals ; Antibodies, Monoclonal ; therapeutic use ; Brain-Derived Neurotrophic Factor ; immunology ; Cell Line, Tumor ; Humans ; Male ; Mice ; Mice, SCID ; Multiple Myeloma ; drug therapy ; metabolism ; pathology ; Neoplasms, Plasma Cell ; drug therapy ; Xenograft Model Antitumor Assays