bluuebunny/crossref_metadata_2025_split huggingface dataset:
Xanthohumol Induces Apoptosis in B-CLL Mediated by Caspase 9 Following Downregulation of bcl-xL and mcl-1.
Authors: Vanhoecke Barbara , Lust Sofie , Janssens Ann , Philippe Jan , Bracke Marc , Offner Fritz 
Published: April 2020

Abstract
Introduction
Xanthohumol is a hop-derived prenylated chalcone with structural similarities to flavopiridol. It has documented impact on breast cancer cell growth and invasiveness in vitro. Based on these observations, lymphocytes from patients with 20 B-CLL were cultured in the presence of xanthohumol in vitro.
Methods
Xanthohumol was dissolved in DMSO and used at concentrations up to 50 μM. B-CLL cells were cultured in RPMI 1640–10% FBS in vitro. Apoptosis, cell cycle analysis and bcl-2 expression was assessed by flow cytometry. PARP, Erk1/2, p38, Akt, JNK, p70S6K, and P-p38, P-Akt, P-JNK, and P-p70S6K, caspase 3, 8 and 9, and bcl-XL and mcl-1 were assessed by Western Blot.
Results
After 24h, xanthohumol induced dose-dependent killing of B-CLL cells at a LD50 (24h) of 24.4 ± 6.6 μM, independently of known adverse prognostic factors such as Ig mutation status, ZAP-70, and cytogenetic abnormalities including 17p-loss of p53. Cell death was associated with PARP cleavage and Annexin V positivity, suggestive of an apoptotic mechanism. Apoptosis was accompanied by cleavage of procaspase-9 but not of procaspase-8. Among bcl-2 family members tested, there was a decrease in bcl-2, bcl-xL and mcl-1 expression. The effects of xanthohumol on the major signal transduction pathways showed no effect on Jnk, Akt, p38 and Erk phosphorylation, but stimulation of p70S6K phosphorylation which could not be inhibited by rapamycin, a specific inhibitor of mTOR.
Conclusion
Xanthohumol has antitumor activity on B-CLL cells in vitro that appears to be independent both of DNA-damage and of Zap-70 effects on signal transduction.
MDM2 Antagonists Activate p53 and Synergize with Genotoxic Drugs To Induce Apoptosis in B-Cell Chronic Lymphocytic Leukemia Cells but Not in T Cells.
Authors: Gil Joan , Coll-Mulet Llorenç , Iglesias-Serret Daniel , Santidrián Antonio F. , Castaño Esther , Campàs Clara , Barragán Montserrat , Cosialls Ana M. , De Frias Mercé , de Sevilla Alberto Fernández , Domingo Alicia , Vassilev Lyubomir T. , Pons Gabriel 
Published: April 2020

Abstract
B-cell chronic lymphocytic leukemia (B-CLL) is characterized by the accumulation of long-lived CD5+ B lymphocytes. Several drugs used currently in the therapy of B-CLL act, at least partially, through activation of the p53 pathway. Recently, non-genotoxic small-molecule activators of p53, the nutlins, have been identified that inhibit p53-MDM2 binding. We have investigated the antitumor potential of nutlin-3 in B-CLL and find that it can effectively activate the p53 pathway and induce apoptosis in cells with wild-type but not mutant p53. The half-maximal effective concentration (EC50) for B-CLL cells was 4.7 ± 1.5 μM (n = 8). Nutlin-3 stabilized p53 and induced several p53 target genes including MDM2, p21, PUMA, Bax, PIG3 and WIG1. At lower concentrations, nutlin-3 synergized with the genotoxic drugs doxorubicin, chlorambucil, and fludarabine but not with acadesine, that induces p53-independent apoptosis. Normal human T cells showed lower sensitivity to nutlin-3 than B-CLL cells and no synergism with the genotoxic drugs. These results suggest that MDM2 antagonists alone or in combination with chemotherapeutic drugs may offer a new treatment option for B-CLL.
The Research of Specific Inhibition of Survivin Expression To Reverse Drug Resistance with RNA Interference Technology.
Authors: Li Juan , Zhao Ying , Luo Shao-kai , Zhang Dian-bao , Haung Bei-hui 
Published: April 2020

Abstract
Objective:Use RNA interference technology to down-regulate the expression of survivin gene in KM3 cells, observe its induction of apoptosis of KM3 cells, and if it can increase chemotherapy sensitivity of KM3 cells to adriamycin.
Methods: SiRNA designed and in-vitro chemosynthesized aiming to survivin was transfected into human myeloma cell line KM3 mediated by liposome. Survivin mRNA transcription and protein expression of KM3 cells were then detected using RT-PCR and Western-blot 24, 48 and 72 hours after transfection, so as to invest the silencing effect. Besides, the apoptosis of the cell line were observed under fluorescence microscopy before and after transfection. Furthermore, cytotoxic analysis was used to compare the sensitivity variation of KM3 cells to adriamycin before and after transfection.
Results: Down-regulations of survivin mRNA transcription and protein expression of KM3 cells could be observed 24 hours after siRNA tranfection, which were (12±2.3)% and (9.4±1.8)% respectively. After 48 hours, they were (61.4±7.9)% and (38.6±6.7)% respectively, while the survivin mRNA transcription and protein expression was increased quickly 72 hours later. Under fluorescence microscopy, the apoptosis rate of KM3 cells transfected with survivin siRNA 48 hours later was 28±7%, which was significantly higher than the control cells. (P&lt;0.05). The IC50 value of KM3 cells to adriamycin decreased from 1.12±0.14uM to 0.21±0.03uM, which means the sensitivity of KM3 cells to adriamycin was 5.3 fold increased.
Conclusions: SiRNA which was designed and synthesized to target survivin could effectively down-regulate survivin mRNA transcription and protein expression in KM3 cells, induce KM3 cells to apoptosis, increase its sensitivity to adriamycin after down-regulation, and effectively reverse drug-resistance of MM cells.
Validation of International Staging System (ISS) for Multiple Myeloma: A Retrospective Analysis of 487 Patients at 8 Brazilian Centers.
Authors: Hungria Vania T.M. , Maiolino Angelo , Martinez Gracia , Colleoni Gisele , Oliveira Luciana , Coelho Erika O.M. , Rocha Lais , Bittencourt Rosane , Quero Adriana , Ribeiro Karina 
Published: April 2020

Abstract
Introduction: The survival of patients with multiple myeloma varies from a few months to more than 10 years. This heterogeneity is related to the characteristics of the myeloma itself and of the host. The identification of the factors which influence the prognosis is very important to predict the result, assist in the choice of the treatment and adequately stratify the patients in clinical studies. Many prognostic factors have been identified in patients with multiple myeloma, such as anemia, renal failure, ß2 microglobulin, albumin and chromossomic alterations. Some authors have combined prognostic factors and proposed various systems of staging. However, none of them have yet substituted the Durie-Salmon staging system. Recently, the International Myeloma Working Group, with the objective of developing a simple and reliable staging system, which can be internationally applied to classify and stratify patients with multiple myeloma, identified 3 risk groups. This new system of staging, the “International Staging System” (ISS), consists of stage I: ß2 microglobulin &lt; 3.5 mg/L plus albumin ≥ 3.5 g/dL (median survival: 62 months); stage II: neither I nor III (median 44 months); stage III: ß2 microglobulin &gt; 5.5 mg/L (median 29 months). This study included sites in North America, Europe and Asia, but the sites in Latin America were not included.
Objective: To validate the ISS in patients with multiple myeloma at Brazilian centers.
Patients and Methods: Four hundred and eighty-seven patients with the diagnosis of multiple myeloma within the period of 1998 to 2004 at Santa Casa de São Paulo, Hospital Universitário Clementino Fraga Filho do Rio de Janeiro, Hospital das Clínicas de São Paulo, Hospital São Paulo, Hospital das Clínicas de Ribeirão Preto, HEMOPE, Hospital Prof. Edgar Santos de Salvador and Hospital de Clínicas de Porto Alegre, with available data on albumin and ß2 microglobulin, were stratified according to the ISS. A total of 339 patients received standard therapy and 148 received high-dose therapy as initial therapy. The survival was estimated using the Kaplan-Meier method with differences in survival examined using the logrank test.
Results: The median age of the patients was 60 years, 52% male and 48% female. In Stage I (n=104), the global median survival was not reached, the survival at 60 months was 60%, in stage II (n=264), the global median survival was 61 months and in stage III (n=119), 19 months (p&lt;0.001).
Conclusion: The new system of staging for multiple myeloma (ISS) is simple, based on variables easy to be applied and was possible to be validated in patients with multiple myeloma in Brazilian centers.
Wnt3 Augments Adhesion Mediated Drug Resistance of Myeloma Cells Via Wnt/RhoA Pathway.
Authors: Chiba Hiroki , Kobune Masayoshi , Kato Kazunori , Nakamura Kiminori , Kawano Yutaka , Takimoto Rishu , Matsunaga Takuya , Kato Junji , Niitsu Yoshiro , Hamada Hirofumi 
Published: April 2020

Abstract
Despite the tremendous effort in developing of conventional chemotherapy and molecular targeting drugs for patients with multiple myeloma (MM), it has been proven difficult to completely abrogate neoplastic cells from bone marrow (BM). Hence, patients with refractory disease still experience poor outcome due to disease progression. Principle obstacle in the treatment of this disease is a chemo-resistance which is mainly caused by the interaction of myeloma cells with BM stromal cells. However, little is known about the molecular mechanism of cell adhesion mediated drug resistance (CAM-DR) in MM. In this study, we focused on relationship between drug resistance and expression of Wnts, the factor regulating the cell adhesion and proliferation, in myeloma cells. To gain insight into involvement of Wnt signaling in CAM-DR, we first screened the expression of Wnt family in myeloma cell lines (RPMI8226, ARH77, KMS-5, HS-sultan and MM1S) by reverse transcription (RT)-polymerase chain reaction (PCR) analysis. Although the mRNAs of Wnt2b, Wnt7a and Wnt10b were variably expressed in some of myeloma cell lines, Wnt3 mRNA was detected in all the myeloma cells examined. RPMI8226, ARH77 and KMS-5 which highly expressed Wnt3 protein, tightly adhered to human BM stromal cells and accumulation of beta-catenin and GTP-bounded RhoA was observed in these myeloma cell lines. This cell adhesion was augmented by addition of Wnt3 containing conditioned medium (CM) and suppressed by Wnt-receptor competitor, secreted Frizzled related protein (sFRP)-1, but not by specific inhibitor of canonical pathway (DKK-1). These results suggest that adhesion of myeloma cells was regulated by non-canonical pathway of Wnt signaling. We further examined whether the Wnt3 mediated adhesion to stromal cells involved in CAM-DR. The drug resistance of ARH-77 for doxorubicin was 1.8 folds enhanced by adhesion to stromal cells in comparison with stroma-free condition. This CAM-DR for doxorubicin was further augmented (2.6 folds) by addition of Wnt3 CM via enhancement of adhesion to stromal cells. Moreover, although the doxorubicin sensitivity of ARH-77 in coculture with stromal cells was significantly reduced by sFRP-1, this effect was not observed in stroma-free culture, indicating that Wnt3-mediated CAM-DR is dependent on attachment with stromal cells. Additionally, CAM-DR was completely restrained by addition of Rho kinase inhibitor Y27632. These results indicate that Wnt3 augments myeloma CAM-DR by enhancement of adhesion to human BM stromal cells via Wnt/RhoA signaling. Thus, Wnt/RhoA signaling pathway could be a promising molecular target to overcome CAM-DR.
Immunoglobulin Free Light Chains at Diagnosis: Predictors of Progression and Survival in Solitary Plasmacytoma of Bone.
Authors: Dingli David , Kyle Robert A. , Rajkumar Vincent S. , Nowakowski Grzegorz S. , Larson Dirk R. , Bida John P. , Gertz Morie A. , Dispenzieri Angela , Melton L. J. , Therneau Terry M. , Katzmann Jerry A. 
Published: April 2020

Abstract
Background: Solitary plasmacytoma of bone (SBP) is a localized collection of monoclonal plasma cells that is potentially curable with local radiation therapy but associated with a high risk of progression to multiple myeloma. We hypothesized that an abnormal immunoglobulin free light (FLC) ratio at diagnosis may be a prognostic indicator of transformation risk.
Methods: We identified a cohort of 133 patients with SBP for whom stored serum taken at the time of diagnosis was available. The diagnosis was ascertained and serum FLC determined in 126 patients.
Results: From this cohort, 48 patients have progressed to myeloma and the median time to progression among those who progressed was 1.9 years. On univariate analysis, age (p&lt;0.001), gender (p=0.035), abnormal FLC ratio at diagnosis (p=0.009) and persistence of serum or urine M-protein after therapy (p=0.0070 were all associated with a shorter overall survival (OS) and time to progression to multiple myeloma.
Progression by Normal FLC(0.26–1.65) Progression by Normal FLC(0.26–1.65)
On multivariate analysis, an abnormal FLC ratio retained its independence in a model that includes age at diagnosis but lost its significance when combined with persistence of the serum or urine M-protein. However, serum or urine M-proteins are not detectable in a significant number of patients with SBP and therefore not informative.
Conclusion: The FLC ratio at the time of diagnosis of SBP is a powerful predictor of risk and a useful aid to management of patients with this condition.
Differential Presence of Truncated Forms of Transcription Factor Yin Yang 1 (YY1) in Bone Marrow of Multiple Myeloma Patients.
Authors: Neshat Mehran S. , Chen Haiming , Gordon Melinda S. , Berenson James R. , Bonavida Benjamin 
Published: April 2020

Abstract
The transcription factor Yin Yang 1 (YY1) regulates cellular differentiation and response to apoptotic stimuli. YY1 exerts its pleiotropic effects through regulation of promoter activity of critical genes, as well as association and direct modulation of stability and function of a subset of proteins. Genes that are regulated by YY1 include those that control the cell cycle, development, differentiation and tumor suppression. For example, it has been reported that YY1 inhibits the proto-oncoprotein c-Myc (Austen, et al., Oncogene, 1998, 17:511) and negatively regulates the tumor suppressor gene p53 (Sui, et al., 2004, Cell 117: 859). Thus, expression and activity of YY1 in tumor cells may be involved in the pathogenesis of disease, as well as controlling response to drug stimuli. YY1 is regulated at transcriptional and post-translational levels in response to intra and extracellular signals. It has been reported that YY1 undergoes proteolytic cleavage. Caspase-dependent N-terminal cleavage of YY1 has been reported in response to physiological (Fas, TNF, L-glutamate) and chemical (staurosporine, etoposide, okadaic acid) death promoting factors. Similar presence of truncated YY1 is observed in in vitro models of skeletal and cardiac muscle differentiation. N-terminal truncated YY1 lacks its transactivation domain, while DNA binding remains unaltered. Hence, YY1 function may be altered by truncated forms. We hypothesized that post-translational processing of YY1 occurs in bone marrow and may be important in tumor progression and response to therapeutic agents. This study thereby aimed to determine whether altered levels and/or forms of YY1 are expressed in the bone marrow of multiple myeloma patients and to identify their potential downstream effectors. YY1 expression in protein lysates of bone marrow aspirates from nine patients was determined by Western blot analysis. Truncated species of YY1 were present in 6/8 samples. In contrast to myeloma bone marrow, one plasma cell leukemia sample showed high levels of YY1 and no truncated forms. Similar high levels of YY1 expression was observed in established tumor xenografts of a plasma cell leukemia tumor. We are presently extending the pool of analyzed normal and cancer harboring tissues and examining potential correlation of YY1 and its altered forms with disease status and prior therapeutic history. Identification and purification of cell populations that generate altered forms of the protein and its effect on expression and function of YY1 interacting proteins are under investigation.
Circulating Nucleic Acids in Multiple Myeloma: Presence of Non-Coding/ Intergenic DNA Reflects Disease Activity.
Authors: Durie Brian G.M. , Jagannath Sundar , Schutz Ekkehard , Urnovitz Howard 
Published: April 2020

Abstract
Circulating nucleotides in plasma have been previously quantified and sequenced in studies involving 65 patients with multiple myeloma (Acta Onc39, 7892000). RT-PCR revealed RNA incorporating a 709-nucleotide sequence (GenBank #AFO18254) with 99.7% homology with a non-coding ALU sequence of a type capable of retro-transposition and disease linked mutation. Levels of this circulating ALU RNA correlated with myeloma disease status. Circulating DNA, devoid of cellular material was also noted as part of these studies. We now describe the molecular analysis of this circulating DNA before, during, and after therapy. Based upon prior findings, direct PCR was performed utilizing primers with a 3′ Alu sequence linked to a variety of 5′ sequences. Detailed studies including serial evaluations were conducted in one patient. 167 total clones were analyzed and categories set to compare under and over-representation in comparison of active myeloma samples versus samples from healthy controls. 96 selected clones fitted into 10 homologs. Homolog #2, which proved to be reflective of myeloma disease activity, incorporated only non-coding DNA regions. One homolog sequence, CNA Clone #2, matched one GenBank entry in the human genome (GenBank #AC020734; chromosome 4), incorporating only repetitive elements and interspersed DNA. The percentage occurrence rates for different types of repeat elements in CNA Clone 2 were: LINEs 25.2%; MaLR 5.5%; HERVs 4.8%; ALUs 4.8%; MERs 3.8%; MiR 2.1%; microsattelites 1.1% and A-T rich regions 1.0%. Using a combination of the 10 homologs containing ALU-attached sequences, distinct patterns of clonal change occurred over several months of follow-up in the patients studied. Initial clones associated with active disease disappeared with response. Transiently, expressed different clones emerged with intervening infection and disappeared with resolution of infection. Electron microscopy has revealed that circulating nucleotides are present in the form of exosomes or membrane vesicles released into the extra-cellular environment upon exocytic fusion of multivesicular endosomes with the cell surface membranes.
Conclusions: Our studies revealed several important aspects. Firstly, specific types of circulating DNA and RNA accompany the presence of active myeloma. Secondly, the predominant types of RNA and DNA are repetitive elements, especially SINEs, LINEs and ALUs. Thirdly, that these repetitive elements are typically linked to other non-coding sequences derived from interspace gene (intergenic) “hotspots”, which may be linked to particular patterns of disease and/ or treatment responses. The serial testing of DNA patterns has the potential for personalized molecular patient monitoring. Further studies are ongoing to assess these findings in a randomized trial setting and in comparison with proteomic and other markers.
Monitoring of Dendritic Cell Numbers in Patients during the Treatment of Multiple Myeloma.
Authors: Kovarova Lucie , Hajek Roman , Svobodnik Adam , Penka Miroslav , Mayer Jiri 
Published: April 2020

Abstract
Objective: In this study, the proportion of dendritic cell (DC) subsets (myeloid DC1 and plasmacytoid DC2), T cells, B cells and NK cells was evaluated in peripheral blood of patients with multiple myeloma (MM) before and during treatment including autologous transplantation. Also control group of healthy volunteers was evaluated.
Methods: Flow cytometric determination of relative cells number in unmanipulated peripheral blood was based on expression of the surface antigen: T cells (CD3/CD4/CD8), B cells (CD19/CD20), NK cells (CD3/CD16/CD56) and DC (CD83/HLA-DR/CD11c and CD83/HLA-DR/CD123).
Results: Significant difference (p&lt;0.01) was found in initial values of CD83+ cells between the group of healthy volunteers (n = 15; mean count of CD83+ cells 0,26 ± 0,15%; ratio DC1/DC2 = 1,54) and the group of patients before treatment (n = 15; 0,15 ± 0,03% CD83+; DC1/DC2 = 4,55). After induction treatment with VAD regimen (vincristine, adriamycin, dexamethasone) in a group of patients was the mean percentage of DC higher (0,18 ± 0,04% CD83+ cells; DC1/DC2 = 4,77) than initial values. Administration of G-CSF again increased the total DC numbers (0,34 ± 0,11%; DC1/DC2 = 2,3) and intermediate levels of DC counts were found in the apheresis products (0,22 ± 0,05%; DC1/DC2 = 1,21). After engraftment there were found the highest relative DC numbers (0,50 ± 0,21%; DC1/DC2 = 1,85) in patients. Within six months after transplantation were achieved pretreatment DC values when compared with DC values of healthy volunteers (p&lt;0,97) (0,24 ± 0,08%; DC1/DC2 = 1,57). Total numbers of T cells did not significantly differ during treatment only the reverse CD4/CD8 ratio was found in majority of patients within six month after the transplantation.
Conclusions: Untreated patients with MM have significant lower relative numbers of peripheral blood DC in comparison with healthy volunteers. The highest number of total DC was found after engraftment. The ratio DC1/DC2 showed relative majority of DC1 subtype and its the lowest value was found in the apheresis products. Normal DC values comparable with DC values of healthy volunteers were found in patients within six months after transplantation together with the reverse CD4/CD8 ratio.