Efficacy of prophylactic antibiotics for the prevention of neutropenic fever in patients with multiple myeloma receiving high-dose cyclophosphamide for stem cell mobilization.

High-dose cyclophosphamide (HD-Cy) (3 g/m2) plus granulocyte colony-stimulating factor (G-CSF) is a very effective regimen for peripheral blood stem cell (PBSC) mobilization. Unfortunately, it is associated with an increased risk of neutropenic fever (NF). We analyzed the effect of NF on PBSC apheresis results and the efficacy of prophylactic antibiotics for the prevention of NF associated with HD-Cy plus G-CSF for PBSC mobilization in patients with newly diagnosed multiple myeloma (MM). First, patients were divided into NF ( +) and NF ( -) groups according to whether they suffered from NF during mobilization. Second, we divided patients into an antibiotic prophylaxis group and a nonantibiotic prophylaxis group according to whether antibiotic prophylaxis was used during the mobilization period. Our study showed that NF( +) patients (n = 44) had lower CD34 + cell dose collection (median 2.60 versus 5.34 × 106/kg, P < 0.001) and slower neutrophil engraftment and platelet engraftment (median 11 versus 10 days, P = 0.002, and median 13 versus 11 days, P = 0.043, respectively) than NF( -) patients (n = 234). Of note, the nonantibiotic prophylaxis group patients (n = 30) had a 26.7% incidence of NF. In the patients receiving antibiotic prophylaxis (n = 227), the incidence was reduced to 9.3% (P = 0.01). The antibiotic prophylaxis patients had higher CD34 + cell collection (median 5.41 versus 2.27 × 106/kg, P < 0.001) and lower hospitalization cost of mobilization ($ median 3108.02 versus 3702.39, p = 0.012). Thus, our results demonstrate that NF is associated with lower CD34 + cell collection and that antibiotic prophylaxis can reduce the incidence of NF and improve stem cell mobilization and collection outcomes, which reduces the hospitalization cost of mobilization.

On-demand plerixafor added to high-dose cyclophosphamide and pegylated recombinant human granulocyte colony-stimulating factor in the mobilization of patients with multiple myeloma: a treatment with high effectiveness, convenient, and affordable cost.

Objective: The combination of high-dose cyclophosphamide (HD-Cy) (3g/m2) plus granulocyte colony-stimulating factor (G-CSF) and on-demand plerixafor (PXF) has been considered an effective mobilization regimen of patients with multiple myeloma(MM). However, the daily multi-injection regimen of G-CSF poses challenges. This study delves into the efficiency and cost implications of a novel approach, using HD-Cy alongside pegylated G-CSF (PEG G-CSF) and on-demand PXF. Unlike G-CSF, which necessitates daily injections, the half-life of PEG G-CSF extended allows for a single injection. Methods: A retrospective analysis was conducted on 350 MM patients, which were categorized based on their mobilization regimens: Cy+PEG G-CSF+/-PXF (n=66), Cy+PEG G-CSF (n=91), Cy+ G-CSF (n=169), and G-CSF+PXF (n=24). Results: Mobilization with Cy+PEG G-CSF+/-PXF(8.79)yielded a notably higher median CD34+ cell count compared to the other regimens: Cy+PEG G-CSF(4.96), Cy+G-CSF (4.65), and G-CSF+PXF (2.99) (P<0.001). The percentage of patients who achieved >6×106/kg CD34+ cells was significantly higher in the Cy+PEG G-CSF+/-PXF group (77.3%) than in the other mobilization regimens: Cy+PEG G-CSF (41.8%), Cy+ G-CSF (37.3%), and G-CSF+PXF (8.3%) (P<0.001). From a cost perspective, the Cy+PEG G-CSF+/-PXF approach was more economical than the G-CSF+PXF strategy but was marginally costlier than the other two methods. A multivariate assessment highlighted that the combination of Cy+PEG G-CSF with on-demand PXF had a superior potential to achieve the desired harvest (6×106/kg) compared to the Cy+PEG G-CSF protocol without PXF. The incremental cost-effectiveness ratio for each 1% increase in the probability of achieving a successful optimal harvest was $ 97.02 per patient. The incidence of neutropenic fever was 3.0% in the Cy+PEG G-CSF+/-PXF group. Conclusion: The combination of on-demand PXF with HD-Cy and PEG G-CSF offers a cost-effective approach with a high mobilization success rate, manageable side effects, and the convenience of fewer injections. It stands as a promising mobilization strategy for MM patients.

Sequential treatment with bortezomib plus dexamethasone followed by autologous hematopoietic stem cell transplantation in patients with multiple myeloma.

BACKGROUND: Whether the sequential treatment with bortezomib plus dexamethasone (BD) followed by autologous hematopoietic stem cell transplantation (ASCT) could extend the overall survival period in multiple myeloma patients is still not clear. Few large case studies about this therapeutics in multiple myeloma were reported in China. Our purpose was to assess the efficacy and adverse effects of sequential treatment with BD chemotherapy and ASCT in patients with multiple myeloma. METHODS: Fifty-three patients with newly diagnosed or relapsed/refractory multiple myeloma received BD as induction therapy before ASCT. Stem-cell mobilization was undertaken with cyclophosphamide 3 - 5 g/m(2) plus granulocyte colony-stimulating factor 300 µg/d. Target yield was 2.0×10(6) CD34(+) cells/kg. Conditioning for ASCT consisted of melphalan 200 mg/m(2). Thalidomide and/or a-interferon was used as post-transplantation maintenance treatment. RESULTS: The BD chemotherapy before transplantation was effective in 86.7% of the 53 patients, including 22.6% with complete remission (CR), 39.6% with near complete remission (nCR), and 24.5% with partial remission (PR). The best effect was achieved after two treatment courses. Most bortezomib-related adverse effects were classes 1 - 2. All patients were successfully mobilized after BD for autologous peripheral blood stem cell transplantation. The ASCT was effective in 96.3% of patients, including 49.1% with CR, 32.1% with nCR, and 15.1% with PR. The CR rate was significantly increased (49.1% vs. 22.6%, P < 0.05) by sequential ASCT. Within 27 (range, 6 - 53) months of follow-up, the efficacy of ASCT was maintained in 29 patients and further enhanced by post-transplantation maintenance treatment in four patients. Eleven patients died after transplantation. Among the patients undergoing BD/ASCT treatment, overall survival (OS) was significantly better in newly diagnosed patients in comparison to relapsed/refractory patients (P = 0.046). CONCLUSIONS: BD chemotherapy can be used as an induction therapy prior to ASCT in patients with multiple myeloma. Its rate of effectiveness is high and it alleviates symptoms quickly without affecting peripheral blood stem cell collection. The majority of adverse effects are mild (tolerable). Sequential BD with ASCT is the preferred option for transplant patients. First-line ASCT could prolong survival of newly diagnosed patients rather than delayed ASCT.

[BAFF level in bone marrow and expression of BAFF receptor on B cells in multiple myeloma patients].

This study was purposed to investigate the B cell-activating factor belonging to the TNF family (BAFF) and a proliferation-inducing ligand (APRIL) levels in bone marrow, and the BAFF receptor expression level on B cells in multiple myeloma (MM) patients, in order to explore the characteristics of B cells in bone marrow of MM patients. MM patients were studied before treatment (newly diagnosed group, 19 patients) and after treatment with improvement (stable group, 17 patients), 10 non-hematologic patients were selected as control (control group). The BAFF receptors (BAFF-R) and transmembrane activator and calcium modulator and cyclophilin ligand interactor (TACI) on B cell (CD19(+)), naive B cell (CD19(+)IgD(+)) and memory B cell (CD19(+)CD27(+)) of bone marrow in all groups were detected by flow cytometry. The BAFF, APRIL level in bone marrow supernatant were tested with ELISA. The results showed that the BAFF-R expression level on CD19(+) cells in newly diagnosed group were higher than that in stable group and control group; there was no significant difference between the BAFF-R expression level on CD19(+)IgD(+) cells in newly diagnosed group and stable group, but BAFF-R expression level on CD19(+)IgD(+) cells in newly diagnosed group was higher than that in control group; the BAFF-R expression level on CD19(+)CD27(+) cells in newly group was higher than that in stable group and control group; there was no significant difference between the BAFF-R expression level on CD19(+) cells, CD19(+)IgD(+) cells or CD19(+)CD27(+) cells in stable group and control group. There was no significant difference among the TACI expression level on CD19(+) cells, CD19(+)IgD(+) cells or CD19(+)CD27(+) cells in newly diagnosed group, stable group and control group. The bone marrow supernatant BAFF level in newly diagnosed group was higher than that in stable group and control group, but there was no significant difference between stable group and control group. There was no significant difference among the bone marrow TACI levels in newly diagnosed group, stable group and control group. It is concluded that both the bone marrow BAFF level and the BAFF-R expression level on CD19(+) cell, CD19(+)IgD(+) cells and CD19(+)CD27(+) cells in MM patients increase, which may help to stimulate B cells, thereby may relate with to MM pathogenesis.

[The efficacy and safety of bortezomib-based induction regimen before autologous hematopoietic stem cell transplantation in patients with multiple myeloma].

OBJECTIVE: To investigate the efficacy and safety of bortezomib-based induction regimen followed by autologous hematopoietic stem cell transplantation (ASCT) in patients with multiple myeloma (MM). METHODS: A retrospective analysis was performed upon clinical data of 62 MM patients who received bortezomib-based induction regimen followed by ASCT from June 2006 to June 2011. All patients were followed up to September 30, 2011. RESULTS: Overall response rate [complete remission (CR) + near complete remission (nCR) + partial remission (PR)], ≥ nCR rate (CR/nCR) and CR rate of post-induction with bortezomib-based regimen were 88.7%, 66.1% and 24.2%, respectively. After ASCT, CR rate and CR/nCR rate were increased to 50.0% and 82.3%, respectively, with significant differences (P = 0.003 and P = 0.032). The median time of neutrophil and platelet engraftment was 12.0 (9 - 43) days and 13.5 (0 - 120) days, respectively. Significances were found in neutrophil and platelet engraftment between MM patients with and without prior exposure to alkylating agents. Furthermore, engraftment of neutrophil and platelet in patients receiving peripheral blood stem cell transplantation were faster than those receiving bone marrow transplantation. No unexpected side effects occurred. The median time of follow-up was 26.5 (7-61) months. The median overall survival (OS) was not reached and the median progression-free survival (PFS) was 30 months. There were significant differences in OS and PFS between patients obtaining CR/nCR and those with ≤ PR before ASCT. CONCLUSIONS: Bortezomib-based induction regimen can improve the efficacy of ASCT in MM patients. The side effects are tolerant. Higher response quality before ASCT can translate to high rates of OS and PFS following high-dose therapy and stem cell transplantation.

Differentiation induction enhances bortezomib efficacy and overcomes drug resistance in multiple myeloma.

AIM: It is of clinical importance to find methods to overcome bortezomib resistance. In the current study, we clarified the relationship between resistance to bortezomib and the differentiation status of myeloma cells, and explored the feasibility of induction of differentiation in overcoming bortezomib resistance in myeloma. METHODS: Cell morphology, immunoglobulin light-chain protein secretion levels, and XBP-1 expression were used to evaluate the differentiation status of myeloma cells. Low dose 2-ME2 alone or in combination with ATRA was used to induce differentiation in myeloma cells. RESULTS: The differentiation status of myeloma cells was related to myeloma sensitivity to bortezomib. After successful induction of differentiation, the myeloma cells were more sensitive to bortezomib with decreased growth and an increased rate of apoptosis. Induction of differentiation increased the proteasome workload in myeloma cells by increasing immunoglobulin secretion, while reducing proteasome capacity by decreasing proteasome activity. The imbalance between increased proteasome workload and decreased proteasome capacity is a possible mechanism by which induction of differentiation overcomes myeloma resistance to bortezomib. CONCLUSION: The current study demonstrated, for the first time, that myeloma differentiation status is associated with myeloma sensitivity to bortezomib and that induction of differentiation can overcome myeloma resistance to bortezomib.

[The clinical features of infection in multiple myeloma undergoing autologous hematopoietic stem cell transplantation].

OBJECTIVE: To explore the clinical features of infection in multiple myeloma (MM) undergoing autologous hematopoietic stem cell transplantation (ASCT). METHODS: Thirty-seven patients with MM undergoing ASCT were retrospectively analyzed for type and time of infection, pathogen, and outcome. RESULTS: Fifty-nine cases of infectious complications occurred in 33 patients (89.2%) after ASCT, with 34 cases (57.6%) of bacterial infections in 30 patients, 15 cases (25.4%) of fungal infections in 12 patients, 4 cases (6.8%) of cytomegalovirus (CMV) infection, 3 cases (5.1%) of herpes zoster virus infection and 3 cases (5.1%) of HBV reactivation. The proportion of bacterial infection, fungal infection and virus infection were 62.8%, 28.6% and 8.6% respectively in the early stage after ASCT, and 50.0%, 20.8% and 29.3% respectively in the median stage. Response to first-line antibiotic therapy was seen in 38 cases (64.4%). Infection-related mortality was 8.1% (3 cases). CONCLUSIONS: The incidence of infection in MM patients undergoing ASCT is high and they are susceptible to all pathogens. It is important to choose the right antifungal agents as quickly as possible to reduce infection-related mortality.

[Maintenance therapies of thalidomine and interferon-α in multiple myeloma].

OBJECTIVE: To explore the efficacies and toxicities in multiple myeloma (MM) patients on the maintenance therapies of thalidomide and interferon-α so as to seek the optimal chemotherapeutic regimen. METHODS: A retrospective analysis was conducted for 57 MM patients on the maintenance therapies of thalidomide and interferon-α after introduction and consolidation. And 56 MM patients without maintenance therapy were enrolled as the control group. RESULTS: The values of progression-free survival (PFS) and overall survival (OS) were significantly longer in the maintenance group and this translated into an improved estimated 3-year PFS of 75.4% (71.8%, 83.3%) versus 23.2% in the control group (P < 0.01). The estimated 4-year OS was higher in the maintenance group [89.5% (89.7%, 88.9%) vs 33.9%, P < 0.01]. No statistically significant differences existed among different maintenance groups in terms of PFS and OS. The administration of maintenance therapy extended both PFS and OS for MM patients of various M-proteins (P < 0.05). However, in the thalidomide group, PFS and OS were extended only in MM of immunoglobulin G (IgG) and immunoglobulin A (IgA) but not in light-chain patients. Furthermore, the MM patients of Durie-Salmon (DS) stages II and III and international staging system (ISS) stages II and III extended PFS and OS through maintenance (P < 0.05). While in those of ISS stage I, the differences were insignificant in terms of PFS and OS between two groups. The results were similar between the thalidomide and control groups. The patients achieving a partial remission (PR) or higher response level benefited from the maintenance therapy in terms of PFS and OS (P < 0.05). In the thalidomide group, the patients with below PR prolonged OS (P = 0.031) but did not achieve a longer PFS (P = 0.091). Both PFS and OS were extended through maintenance therapy after either stem cell transplantation or consolidation chemotherapies (P < 0.05). There was no significant difference in terms of PFS and OS between MM patients without maintenance therapy after transplantation and those without transplantation. The adverse effects of thalidomide, milder than those of interferon-α, could be tolerated in most patients. The incidence and severity of adverse effects showed no significant difference between the combination maintenance and single agent therapies. CONCLUSION: The maintenance therapies of thalidomide and interferon-α could improve the profiles of PFS and OS in MM patients. And there was no significant difference between them in terms of PFS and OS. However, the maintenance therapy of thalidomide is a better option due to its convenient application, milder adverse effects, reasonable cost and better efficacies in MM patients not achieving PR or receiving induction therapy without bortezomib or without transplantation.

[Combination of bortezomib and dexamethasone for newly diagnosed multiple myeloma].

OBJECTIVE: To analyzed retrospectively two groups of patients with newly diagnosed multiple myeloma (MM) receiving bortezomib and dexamethasone (VD) regimen and vincristine combined with pirarubicin and dexamethasone and melphalan(VADM) regimen. METHODS: Twenty-four patients were enrolled in a group of VD, receiving bortezomib 1.3mg/m(2) on days 1, 4, 8, 11 and dexamethasone 20mg on days 1-4 intravenously of every 21-day cycle. EBMT Standard was used to evaluate the efficacy and NCI-CTC V3.0 was used to decide the adverse effect. Thirty matched patients with newly diagnosed MM who received VADM were used as control group, receiving vincristine 0.4 mg/d and pirarubicin 9 mgxm(-2)xd(-1) and dexamethasone 20 mg/d and melphalan 12 mg/d on days 1 - 4 intravenously of every 28 day cycle. RESULTS: With a median follow-up of 10.5 months in VD group, there were 87.5% patients (21/24) responded, including 12 cases (50.0%) of complete remission (CR) or near complete remission (nCR). The total response rate (RR) was 76.7% in VADM group, with no significant difference in VD group (P = 0.483). CR + nCR rate was significantly higher in VD group than in VADM group (10%) (P = 0.001). RR and CR + nCR of light chain patients in VD group were significantly higher than in VADM group (P = 0.025 and 0.040, respectively). The median time to response and to best response were significantly shorter in VD group than in VADM group. In VD group, the RR of 8 patients with renal dysfunction was 87.5%, and that of 16 with normal renal function was 75% (P = 0.631). There was no significant difference in adverse effects between patient with renal dysfunction and normal function (P > 0.05). The main adverse effects in VD group were fatigue (66.7%), diarrhea (58.3%), peripheral neuropathy (54.2%), thrombocytopenia (29.2%), infection (29.2%), fever (25.0%) and constipation (25.0%). Most of the adverse effects were mild (grade 1 - 2) and could be relieved by symptomatic treatments. The most common adverse event in VADM group was neutropenia (83.8%), infection (35.5%), vomiting (35.5%), loss of hair (32.5%) and thrombocytopenia (16.2%). CONCLUSION: VD has higher CR + nCR rate compared with VADM and can be tolerant in most patients. VD is safe in patients with renal inadequacy.

[The clinical features and risk factors for invasive fungal infection in multiple myeloma.].

OBJECTIVE: To study the clinical features and risk factors of invasive fungal infection (IFI) in multiple myeloma (MM). METHODS: Three hundred and fifty-seven cases of MM were retrospectively analyzed for IFI, clinical features, complicating diseases, treatment of fungus and side effect of anti-fungal drugs. RESULTS: Forty-four cases (12.3%) of IFI were diagnosed. Three of them were diagnosed definitely, 8 clinically and 33 probably. Ten cases incurred IFI in the induction therapy, 4 in platform, 27 in progress and 3 in the treatment with autologous stem cell transplantation. The lung was the commonest site of infection (50.0%). The total effective rates of amphotericin B liposome, voriconazole, itraconazole, caspofungin and fluconazol were 83.3%, 75.0%, 78.9%, 75.0% and 57.1% respectively (P = 0.493). In a multivariate analysis, independent factors significantly associated with IFI were diabetes (P = 0.035, OR 2.527, 95%CI 1.005 - 6.052), dialysis (P = 0.022, OR 2.768, 95%CI 1.161 - 6.600), persistent agranulocytosis (P = 0.019, OR 3.215, 95%CI 1.200 - 7.407), broad-spectrum antibiotic therapy (P = 0.009, OR 3.350, 95%CI 1.353 - 8.295) and fludarabine treatment (P = 0.001, OR 4.669, 95%CI 1.813 - 12.023). CONCLUSIONS: Patients with MM are in high risk of IFI. The lung is the commonest site of infection. The therapeutic effect was similar with itraconazole, voriconazole, caspofungin and amphotericin B liposome in MM patients with complicating IFI. The risk factors for IFI in MM were diabetes, dialysis, persistent agranulocytosis and the use of broad-spectrum antibiotics and fludarabine.

[The clinical features, chemotherapy responses and survival of 223 patients with newly diagnosed multiple myeloma].

OBJECTIVE: To explore the proportion, clinical and laboratory features, chemotherapy responses and long term survival of different kinds of newly diagnosed multiple myeloma (MM) in China. METHODS: The clinical data of 223 cases of newly diagnosed MM patients were gathered in the First Affiliated Hospital of Sun Yat-sen University from Jan, 2000 to Seb, 2007 were retrospectively analyzed. RESULTS: The proportions of each kind of MM including IgG, IgA, light chain, IgD, IgM and biclonal MM were 48.0%, 20.6%, 25.6%, 4.0%, 0.9% and 0.9% respectively. No IgE and nonsecretory myeloma was found. The median age of onset was 58 years, of which that of the IgA type was the oldest one and the light chain type was the youngest (P = 0.004). Bone pain, renal insufficiency, and anemia were the most common symptoms which accounted for 67.7%, 61.0% and 45.3% respectively. The incidences of renal inadequacy, hypercalcemia and pathological fracture in light chain type were higher than those in IgG and IgA types. Besides, no M protein were found in serum protein electrophoresis and no elevation of total globulin. The clinical features of IgD type were similar to that of the light-chain type. The total chemotherapy efficacy rate of 89 patients who were treated with more than 3 cycles in our hospital is 61.8%, which has no difference in all types. Median overall survival of the 89 patients was 33.0 months. CONCLUSION: IgG is the most common type in MM. Bone pain, anemia, hypercalcemia and renal insufficiency are common symptoms. Immunofixation electrophoresis should be performed routinely to avoid missed diagnosis of light-chain and IgD types of MM.

[Imatinib induces c-kit positive myeloma cells apoptosis].

OBJECTIVE: To explore the influence of Imatinib on multiple myeloma cells expressing c-kit in vitro and its mechanism. METHODS: KM3 cells were treated with Imatinib at different concentrations, and cell growth index were evaluated by XTT assay, cell cycle by flow cytometry, apoptosis by Annexin V/ PI and DNA ladder, and change in protein level by Western blot. RESULTS: Imatinib inhibited proliferation of KM3 cells at concentrations more than 0.25 micromol/L in a dose-dependent manner, and the 48 h IC50 was 0.33 micromol/L (P < 0.01). Imatinib arrested cell in C0/G1 phase. Annexin V/PI staining and DNA ladder indicated that Imatinib had a substantial effect on inducing apoptosis of KM3 cells in a dose-dependent manner and induced pro-caspase-3 and poly ADP-ribose polymerase (PARP) cleaved. Imatinib inhibited expression of c-kit and provoked a decrease of IL-6 induced c-kit phosphorylation in vitro. CONCLUSION: Imatinib inhibits KM3 cells proliferation and induces the cells apoptosis by inhibiting c-kit signalling transduction.

[Efficacy of bortezomib combined dexamethasone in 24 patients with multiple myeloma].

BACKGROUND & OBJECTIVE: Bortezomib, a potent and reversible proteasome inhibitor, induces apoptosis of myeloma cells, resulting in durable responses in patients with multiple myeloma (MM). This study was to explore the medical effects and side effects of bortezomib combined dexamethasone in treating newly diagnosed and relapsed or refractory MM, and evaluate the safety of this regimen in the patients with renal impairment. METHODS: Twenty-four MM patients were treated with bortezomib and dexamethasone in a 21-day cycle. The patients received a median of 3 cycles (range, 1-8 cycles) of the treatment. Response to bortezomib was evaluated according to the criteria of the European Group for Blood and Marrow Transplantation (EBMT); adverse events were graded according to the National Cancer Institute Common Toxicity Criteria. RESULTS: During the follow-up with a median of 4 months, 19 (79.2%) patients responded to the treatment. The complete remission (CR) rate was significantly higher in the patients of light-chain type than in those of non-light-chain type (57.1% vs. 5.9%, P=0.014). The response rates of the patients with and without renal impairment were similar (100% vs. 70.6%, P=0.272), and the renal functions were ameliorated in the patients with renal impairment during chemotherapy. Grade III-IV adverse events, including leucocytopenia (8.3%), thrombocytopenia (33.3%), diarrhea (8.3%) and debility (4.2%), could be relieved by symptomatic treatment or delayed chemotherapy. CONCLUSIONS: The combination of bortezomib and dexamethasone shows obvious effects on MM, especially in the patients with light-chain type. The adverse events can be tolerant in most patients, and this regimen is also safe in the patients with renal impairment.

[Expression and clinical significance of beta-catenin in multiple myeloma].

BACKGROUND & OBJECTIVE: beta-catenin is the pivotal regulator in Wnt pathway and in charge of cellular adhesion and signal conduction. Overexpression of beta-catenin has been observed in various human tumors. This study was to investigate the expression and clinical significance of beta-catenin in multiple myeloma (MM). METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to detect mRNA and protein expression of beta-catenin in bone marrow samples from 12 newly diagnosed MM patients, 14 relapsed/refractory MM patients, and 11 healthy donors. The clinical data and treatment outcomes of the MM patient were also analyzed. RESULTS: The positive rate and the expression level of beta-catenin mRNA were significantly lower in healthy donors than in newly diagnosed patients and relapsed/refractory patients (27.3% vs. 75.0% and 100%, P=5.01e(-4); 0.35+/-0.17 vs. 0.72+/-0.11 and 0.85+/-0.16, P=5.88e(-5)); the mRNA level of beta-catenin was significantly lower in newly diagnosed patients than in relapsed/refractory patients (P=0.045). beta-catenin protein was not detected in healthy donors; while the positive rate and the expression level of beta-catenin protein were significantly lower in newly diagnosed patients than in relapsed/refractory patients (50.0% vs. 85.7%, 0.32+/-0.11 vs. 0.21+/-0.08, P=0.039). In the 10 newly diagnosed MM patients with evaluable treatment outcomes, the positive rate of beta-catenin protein was significant lower in the 7 patients without response than in the 3 patients showed response (14.3% vs. 100%, P=0.033). The positive rate of beta-catenin protein was significant higher in the 16 Durie/Salmon stage III patients than in the 10 stage II patients (87.5% vs. 40.0%, P=0.026), and significant higher in ISS stage III patients than in stage I and II patients (100% vs. 45.5% and 33.3%, P=0.006, P=0.032). The protein level of beta-catenin was positively correlated to serum levels of beta2-MG (r=0.688, P=0.002) and lactate dehydrogenase (LDH) (r=0.502, P=0.034). CONCLUSION: The expression of beta-catenin is related with treatment outcome, Durin-Salmon stage and ISS stage of MM, and is positively correlated to serum levels of LDH and beta2-MG.

[Effect of silencing survivin gene on sensitivity of myeloma cells to adriamycin].

BACKGROUND & OBJECTIVE: Survivin, a member of the inhibitors of apoptosis protein family (IAPs), is overexpressed in many cancers, but not in normally differentiated adult tissues. It is known to be involved in poor prognosis and resistance to chemotherapy and radiotherapy in many cancers. This study was designed to use RNA interference technique to down-regulate the expression of survivin gene in human myeloma cell line KM3, observe its effects on apoptosis of KM3 cells and sensitivity of KM3 cells to adriamycin (ADM). METHODS: Small interfering RNA (siRNA) targeting survivin mRNA was designed and in vitro chemosynthesized, and transfected into KM3 cells. Survivin mRNA and protein levels in KM3 cells were detected using reverse transcription-polymerase chain reaction (RT-PCR) and Western blot 24, 48, and 72 h after transfection. The apoptosis of KM3 cells was observed under fluorescent microscope before and after transfection. The sensitivity of KM3 cells to ADM before and after transfection was assessed. RESULTS: The mRNA level of survivin was down-regulated by (47.0+/-4.3)%, (81.4+/-6.2)%, and (49.1+/-7.9)% of control at 24, 48, and 72 h after siRNA transfection, respectively; while the protein level of survivin was down-regulated by (39.6+/-3.8)%, (56.4+/-6.7)%, and (68.2+/-5.1)% of control, respectively. Under fluorescent microscope, the apoptosis rate of KM3 cells was (28+/-7)% at 48 h after transfection of survivin siRNA, which was significantly higher than that of control cells after transfection of negative siRNA (P<0.05). The 50% inhibition concentration (IC(50)) of ADM to KM3 cells was decreased from (1.12+/-0.14) micromol/L to (0.31+/-0.03) micromol/L. CONCLUSION: Down-regulation of survivin expression in KM3 cells by siRNA could effectively induce apoptosis of KM3 cells and increase their sensitivity to ADM.

[Expression and clinical significance of survivin in bone marrow cells of multiple myeloma patients].

BACKGROUND & OBJECTIVE: Survivin, a member of the inhibitors of apoptosis protein family (IAPs), is overexpressed in many cancers, but not in normal differentiated adult tissues. It is related to poor prognosis and resistance to chemotherapy and radiotherapy in many cancers. This study was designed to detect the expression of Survivin in bone marrow of healthy people, newly diagnosed and relapsed/refractory multiple myeloma (MM) patients, and MM cell line KM3, to explore its correlation to drug resistance and prognosis of MM patients. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to detect mRNA and protein expression of Survivin in bone marrow samples of 17 newly diagnosed MM patients, 12 relapsed /refractory MM patients, and 9 healthy people, and myeloma cell line KM3. RESULTS: KM3 cells expressed Survivin strongly. The positive rates and the expression levels of Survivin mRNA were significantly lower in healthy people than in newly diagnosed patients and relapsed/refractory patients (22.2 % vs. 70.5 % and 83.3 %, 0.06+/-0.04 vs. 0.31+/-0.14 and 0.69+/-0.24, P<0.01), and were significantly lower in newly diagnosed patients than in relapsed/refractory patients (P<0.05). Survivin protein were not detected in all of the healthy people; while the positive rates of Survivin were 41.4% in newly diagnosed patients and 58.3% in relapsed/refractory patients (P>0.05), and its expression levels was significantly higher in relapsed/refractory patients than in newly diagnosed patients (0.21+/-0.12 vs. 0.11+/-0.07, P<0.05). Among the 7 Survivin-positive newly diagnosed patients, 4 achieved partial remission (PR), 1 achieved MR, and 2 showed no change (NC); among the 10 Survivin-negative newly diagnosed patients, 2 achieved complete remission (CR), 4 achieved PR, 2 achieved MR, and 2 showed NC. The effective (CR+PR+MR) rate was slightly higher in Survivin-negative patients than in Survivin-positive patients (80.0% vs. 71.4%). CONCLUSION: High expression of Survivin may be relates to drug resistance in some patients. Since Survivin only expressed in bone marrow cells of multiple myeloma, but not in healthy people, it can be a new target to reverse drug resistance in myeloma cells.