Burkitt's Lymphoma Cell Research Articles

Therapeutic Effects of a Novel Fusion of ALT-803, an IL-15 Superagonist, with 4 Single-Chains of Anti-CD20 Antibody (2B8T2M) in Combination with Expanded Natural Killer Cells Against Rituximab Sensitive and Resistant Burkitt Lymphoma (BL)

Background:The prognosis of patients with relapsed or progressed B-cell (CD20+) non-Hodgkin lymphoma (B-NHL), including Burkitt lymphoma (BL), is dismal due to chemo-radiotherapy resistance (Cairo et al, JCO, 2012, Cairo et al, BJH, 2016). Rituximab has been widely used in frontline treatment of B-NHL, however, some patients retreated with rituximab relapse which limit patient treatment options (Goldman/Cairo, Leukemia, 2013). Several strategies for overcoming rituximab-resistance are currently being evaluated, including engineering immune cells with chimeric antigen receptors (Chu/Cairo et al, Can Imm Res 2015), as well as second-generation engineered anti-CD20 antibodies (Tiwari/Cairo et al BJH 2015). Nature Killer (NK) cells are bone marrow-derived cytotoxic lymphocytes that play a major role in the rejection of tumors. A variety of activating and inhibitory receptors on the NK cell surface are engaged to regulate NK cell activities. However, NK therapy is limited by several factors, including small numbers of active NK cells in unmodified peripheral blood, lack of tumor targeting specificity, and multiple mechanisms of tumor escape of NK cell immunosurveillance. Our group has successfully expanded functional and active peripheral blood NK cells (exPBNK) with irradiated feeder cells (Chu/Cairo, et al, Can Imm Res 2015). ALT-803 (Altor BioScience Corporation) is a superagonist of an IL-15 variant bound to an IL-15RαSu-Fc fusion with enhanced IL-15 biological activity (Zhu et al . 2009 J Immunol), longer half-life and increased potency (Han, et al . Cytokine . 2011). Recently, a new therapeutic molecule 2B8T2M was generated by fusing ALT-803 to four single-chains of the tumor-targeting monoclonal antibody, rituximab (Liu/Wong, et al, JBC, 2016). 2B8T2M displayed tri-specific binding activity through its recognition of the CD20 molecule on tumor cells, activated NK cells to enhance antibody-dependent cellular cytotoxicity (ADCC), and induced apoptosis of B-lymphoma cells (Liu/Wong, et al, JBC, 2016).Objective: To examine if 2B8T2M significantly enhances the cytotoxicity of exPBNK against rituximab-sensitive and -resistant BL cells.Method: PBMCs were expanded with lethally irradiated K562-mbIL21-41BBL cells (Denman/Dean Lee, PLoS One, 2012). CD56+CD3- exPBNK cells were isolated using Miltenyi NK cell isolation kit. ALT-803 and 2B8T2M were generously provided by Altor BioScience Corporation. NK receptors expression and cytotoxicity were examined as we previous described (Chu/Cairo, et al, Can Imm Res 2015). IFN-γ and granzyme B levels were examined by standard enzyme-linked immunosorbent assays. Equal doses of Rituximab, ALT-803, Rituximab+ALT-803, obinutuzumab (obinu) were used for comparison. IgG was used as controls. Rituximab-sensitive and -resistant BL cells Raji, Raji-2R and Raji-4RH, were used as target cells.Results: Both ALT-803 and 2B8T2M at equal doses significantly enhanced NK activating receptors such as NKG2D, NKp30, NKp44, and CD16 on exPBNK cells compared to IgG controls.2B8T2M significantly enhanced exPBNK cytotoxicity against rituximab-sensitive Raji cells compared to the controls IgG, Rituximab, ALT-803, Rituximab+ALT-803, obinu (2B8T2M vs IgG vs ALT803 vs Rituximab vs ALT803+Rituximab vs obinu=98.34%+0.24% vs 56.5+1.83% vs 62.8+1.1% vs 69.52+1.79% vs 81.85+1.83% vs 81.23+1.8%, p<0.001, E:T=1:1). 2B8T2M also significantly enhanced exPBNK cytotoxicity against rituximab-resistant Raji-2R cells (2B8T2M vs IgG vs ALT803 vs Rituximab vs ALT803+Rituximab vs obinu=90.7%+1.36% vs 20.47+7.7% vs 48.18+8.07% vs 37.44+3.63% vs 51.02+1.42% vs 65.51+3.03%, p<0.001, E:T=1:1) and resistant Raji-4RH cells (p<0.001, E:T=1:1) (Fig.1). Furthermore, we confirmed the enhanced cytotoxicity by measuring IFN-γ and granzyme B production. 2B8T2M significantly enhanced IFN-γ and granzyme B production from exPBNK against rituximab-sensitive Raji and -resistant Raji-2R and Raji-4RH compared to IgG (p<0.001), Rituximab (p<0.001), ALT-803 (p<0.001), Rituximab+ALT-803 (p<0.001), and obinutuzumab (p<0.001).Conclusions: 2B8T2M significantly enhanced exPBNK activating receptor expression and in vitro cytotoxicity against rituximab-sensitive and -resistant BL cells. The in vivo functions of 2B8T2M with exPBNK against rituximab-sensitive and -resistant BL cells using humanized NSG models are under investigation. [Display omitted] DisclosuresAlter:Altor BioScience: Employment, Equity Ownership. Jeng:Altor Bioscience: Employment, Equity Ownership. Rhode:Altor BioScience: Employment, Equity Ownership. Lee:Intellia Therapeutics, Inc: Consultancy; Courier Therapeutics, Inc: Membership on an entity's Board of Directors or advisory committees; Miltenyi: Speakers Bureau; Cyto-Sen Therapeutics, Inc: Other: Founder, Vice President, Medical Director. Wong:Altor BioScience: Employment, Equity Ownership. Cairo:Jazz Pharmaceuticals: Speakers Bureau.

RNA-guided gene editing of the murine gammaherpesvirus 68 genome reduces infectious virus production.

Epstein-Barr virus (EBV) and Kaposi sarcoma herpesvirus (KSHV) are cancer-causing viruses that establish lifelong infections in humans. Gene editing using the Cas9-guideRNA (gRNA) CRISPR system has been applied to decrease the latent load of EBV in human Burkitt lymphoma cells. Validating the efficacy of Cas9-gRNA system in eradicating infection in vivo without off-target effects to the host genome will require animal model systems. To this end, we evaluated a series of gRNAs against individual genes and functional genomic elements of murine gammaherpesvirus 68 (MHV68) that are both conserved with KSHV and important for the establishment of latency or reactivation from latency in the host. gRNA sequences against ORF50, ORF72 and ORF73 led to insertion, deletion and substitution mutations in these target regions of the genome in cell culture. Murine NIH3T3 fibroblast cells that stably express Cas9 and gRNAs to ORF50 were most resistant to replication upon de novo infection. Latent murine A20 B cell lines that stably express Cas9 and gRNAs against MHV68 were reduced in their reactivation by approximately 50%, regardless of the viral gene target. Lastly, co-transfection of HEK293T cells with the vector expressing the Cas9-MHV68 gRNA components along with the viral genome provided a rapid read-out of gene editing and biological impact. Combinatorial, multiplex MHV68 gRNA transfections in HEK293T cells led to near complete ablation of infectious particle production. Our findings indicate that Cas9-gRNA editing of the murine gammaherpesvirus genome has a deleterious impact on productive replication in three independent infection systems.

Enhanced electrochemiluminescence ratiometric cytosensing based on surface plasmon resonance of Au nanoparticles and nanosucculent films

Ramos cells are human Burkitt's lymphoma cells, which are a kind of cancer cells to facilitate the monitoring of the relevant biological processes of cancers. Sensitive and accurate detection of Ramos cells using emerging ratiometric ECL biosensing technology shows increasing importance, however, the target analytes of current ratiometric ECL biosensors are mainly limited to DNA/RNA or proteins. In this study, we proposed a dual-potential ratiometric sensing strategy for the electrochemiluminescence detection of Ramos cells based on two types of electrochemiluminescence (ECL)-responding molecular. Au nanosucculent films (AuNFs) were electrodeposited on the fluorine doped tin oxide (FTO) electrode to increase the effective area of the electrode for more efficient assembly of DNA and effectively improving the conductivity of the sensing interfaces. In the presence of Ramos cells, aptamers capped with Au@luminol would conjugate with Ramos cells and then remove from AuNFs, accompanying the decrease of ECL signal from Au@luminol. Then, Au–DNA was captured and alternately hybridized with DNA-modified CdS nanocrystals (NCs) on the surface of AuNFs with the formation of a super reticulate structure. The reticulate structure not only raised another identified ECL signal from CdS NCs but also greatly promoted its ECL intensity from the surface plasmon resonance originating from Au NPs. The value of log (ECLCdS/ECLluminol) and the logarithm of the number of cells exhibit considerable linear relation ranging from 80 to 8 × 105 cells mL−1 with a low detection limit of 20 cells mL−1 (S/N = 3). The selectivity and specificity of this dual-potential ECL sensor showed good performance and indicated considerable promise in avoiding false-positive results in detection.

Smac mimetics and TRAIL cooperate to induce MLKL-dependent necroptosis in Burkitt's lymphoma cell lines

Burkitt's lymphoma (BL) is a highly aggressive form of B-cell non-Hodgkin's lymphoma. The clinical outcome in children with BL has improved over the last years but the prognosis for adults is still poor, highlighting the need for novel treatment strategies. Here, we report that the combinational treatment with the Smac mimetic BV6 and TRAIL triggers necroptosis in BL when caspases are blocked by zVAD.fmk (TBZ treatment). The sensitivity of BL cells to TBZ correlates with MLKL expression. We demonstrate that necroptotic signaling critically depends on MLKL, since siRNA-induced knockdown and CRISPR/Cas9-mediated knockout of MLKL profoundly protect BL cells from TBZ-induced necroptosis. Conversely, MLKL overexpression in cell lines expressing low levels of MLKL leads to necroptosis induction, which can be rescued by pharmacological inhibitors, highlighting the important role of MLKL for necroptosis execution. Importantly, the methylation status analysis of the MLKL promoter reveals a correlation between methylation and MLKL expression. Thus, MLKL is epigenetically regulated in BL and might serve as a prognostic marker for treatment success of necroptosis-based therapies. These findings have crucial implications for the development of new treatment options for BL.

Dendrosomal nano-curcumin reduces VEGF gene expression and with increasing cell apoptosis has an inhibitory effect on the Burkitt lymphoma cell line.

Dendrosomal nano-curcumin reduces VEGF gene expression and with increasing cell apoptosis has an inhibitory effect on the Burkitt lymphoma cell line.

Construction and Characterization of 1F5 Chimeric Anti-CD20 Monoclonal Antibodies: an Efficient Splicing by Overlap Extension-polymerase Chain Reaction Technique

Despite the unparalleled success of anti-CD20-targeted immunotherapy, the currently available mAbs are not sufficiently efficacious in the treatment of lymphoma. 1F5 is one of a panel of anti-CD20 mAbs that was used in the B-cell lymphoma serotherapy. Despite the efficacy of murine 1F5 mAbs in lymphoma patients, the 1F5 chimeric antibodies with human effector functionality are yet to be approved and widely used in the treatment of lymphoma. In this study, the conversion of 1F5 mAb from mouse IgG2a to human-mouse chimeric IgG1 was achieved and the chimeric antibody was partially characterized. We constructed the 1F5 chimeric mouse-human anti-CD20 antibody genes using an efficient Splicing by overlap extension-polymerase chain reaction (SOE-PCR) technique and cloned the chimeric heavy and light genes in pBudCE4.1 mammalian expression vector, followed by purification of the expressed chimeric 1F5 mAbs using affinity chromatography. Our investigation also included the biological properties of purified chimeric antibodies. The generated 1F5 chimeric mAbs mediate complement-dependent cytotoxicity (CDC) and antibody-dependent cellular cytotoxicity (ADCC) against Raji and Daudi Burkitt's lymphoma cell lines, which were comparable with rituximab and exhibit superior reduction in cell viability in vitro, compared to rituximab. The current study indicated that the generated chimeric 1F5 mAbs has potential CDC and ADCC activity which was comparable with rituximab whereas it exhibits a superior reduction in cell viability, compared to rituximab. Our work contributes to future studies involving in vivo biological functions and the application of the 1F5 chimeric antibody.

B Cell Receptor-Responsive miR-141 Enhances Epstein-Barr Virus Lytic Cycle via FOXO3 Inhibition.

ABSTRACTAntigen recognition by the B cell receptor (BCR) is a physiological trigger for reactivation of Epstein-Barr virus (EBV) and can be recapitulated in vitro by cross-linking of surface immunoglobulins. Previously, we identified a subset of EBV microRNAs (miRNAs) that attenuate BCR signal transduction and subsequently dampen lytic reactivation in B cells. The roles of host miRNAs in the EBV lytic cycle are not completely understood. Here, we profiled the small RNAs in reactivated Burkitt lymphoma cells and identified several miRNAs, such as miR-141, that are induced upon BCR cross-linking. Notably, EBV encodes a viral miRNA, miR-BART9, with sequence homology to miR-141. To better understand the functions of these two miRNAs, we examined their molecular targets and experimentally validated multiple candidates commonly regulated by both miRNAs. Targets included B cell transcription factors and known regulators of EBV immediate-early genes, leading us to hypothesize that these miRNAs modulate kinetics of the lytic cascade in B cells. Through functional assays, we identified roles for miR-141 and EBV miR-BART9 and one specific target, FOXO3, in progression of the lytic cycle. Our data support a model whereby EBV exploits BCR-responsive miR-141 and further mimics activity of this miRNA family via a viral miRNA to promote productive lytic replication.IMPORTANCE EBV is a human pathogen associated with several malignancies. A key aspect of lifelong virus persistence is the ability to switch between latent and lytic replication modes. The mechanisms governing latency, reactivation, and progression of the lytic cycle are only partly understood. This study reveals that specific miRNAs can act to support the EBV lytic phase following BCR-mediated reactivation triggers. Furthermore, this study identifies a role for FOXO3, commonly suppressed by both host and viral miRNAs, in modulating progression of the EBV lytic cycle.

Effects of Emodin Derivative on Cell Cycle, Apoptosis and NF-κB Pathway in Burkitt Lymphoma Cells

To investigate the effect and mechanism of a novel emodin derivative YX-18 on Burkitt lymphoma (BL) cells. MTT assay was used to detect the effect of YX-18 on the proliferation of BL cell lines CA46 and Raji. Annexin V-PE/7-AAD double staining assay was used for detecting the effect of YX-18 on the apoptosis of CA46 and Raji cells. PI/RNase staining was used to test the effect of YX-18 on CA46 and Raji cell cycle. JC-1 method was used to measure the changes of mitochondrial membrane potential after YX-18 treatment, and DAPI staining was used to detect the morphology of apoptotic cells. Western blot was used to analyze the distribution changes of NF-κB pathway protein (P65, P-P65, IκB, P-IκB) in the cytoplasm and cell nucleus, and also the expression changes of cyclin-related protein P21, CDK2, P-CDK2, Cycling D1, Cycling E1, and the apoptosis-related protein Caspase-3, Caspase-8, Caspase-9 and the proliferation-related protein C-MYC, BCL-2 by YX-18. Real-time fluorescence-quantitative PCR was used to evaluate the effects of YX-18 on mRNA levels of C-MYC and Ki-67 genes in CA46 and Raji cells, and EBNA-1 and EBER genes of EBV in Raji (EBV+) cells. Novel Emodin derivative YX-18 could effectively inhibit the proliferation of BL cell lines CA46 and Raji, showing a time-dependent effect (24, 48 and 72 h: rCA46=0.89, 0.75, 0.75, rRaji=0.87, 0.73, 0.64). IC50 of CA46 cells and Raji cells treated with YX-18 for 24 h was 1.77±0.04 µmol/L and 1.97±0.22µmol/L, respectively. CA46 cells and Raji cells were treated with YX-18 at concentration of 2.0 and 4.0 µmol/L for 24 h. Compared with the control group, both strains of cells showed a very significant apoptosis at the concentration of 2.0 and 4.0 µmol/L (P<0.01), showing a concentration-dependent effect (rCA46=0.99, rRaji=0.92). Moreover, the cleavaged Caspase-3, 8 and 9 proteins were activated by YX-18 into verious degrees in both two cell lines. Both the two cell lines displayed by YX-18 cell cycle arrest at G0/G1 phase (P<0.01) after exposed to YX-18 for 24 hours at the concentration of 1.0, 2.0 µmol/L in CA46 cells and at 0.5 and 1 µmol/L in Raji cells, respectively. YX-18 decreased expression level of cyclin D1, cyclin E1, CDK2, p-cdk2 proteins and increased p21Waf1/Cip1 level in CA46 and Raji cells. YX-18 significantly declined mitochondrial membrane potential in both cells at the concentration of 2.0 and 4.0 µmol/l (P<0.01) with concentration-dependent manner (rCA46=-0.96, rRaji=-0.99). Western blot tests indicated that YX-18 down-regulated nucleus P65 and intracellular cytoplasm P65, P-IκB, P-P65 protein, and upregulated intracellular IκB level with dose-dependent manner. Meanwhile, the expression level of the cell proliferation-related molecules C-MYC and BCL-2 was decreased significantly. YX-18 suppressed mRNA levels of C-MYC and Ki-67 in both cell lines, and EBNA-1 in EBV-positive Raji cells in a concentration-dependent way. The novel emodin derivative YX-18 can significantly inhibit the proliferation of Burkitt lymphoma cells, and induce the cell apoptosis and cycle arrest. The inhibitory effect of YX-18 on the proliferation of Burkitt lymphoma cells may be related with the effect of Caspase apoptosis pathway, the proliferation and apoptosis-related molecules, such as C-MYC and Ki-67, and also to the inhibition of NF-κB pathway.

Cell Function Research of β-Trefoil Lectins from Mytilidae

Two novel β-trefoil lectins, MytiLec-1 and SeviL were found from mussels in the coast of Yokohama and Nagasaki. MytiLec-1 was purified from gill and mantle of Mytilus galloprovincialis. It was consisted of 149 amino acid residues and there was no similarity with any other proteins when it was discovered. We advocate for this "Mytilectin" as a new protein family because of their novelty of its primary structure and homologues were also found in other mussels. Glycan array analysis revealed that MytiLec-1 specifically bound to the Gb3 and Gb4 glycan which contained the α-galactoside. MytiLec-1 caused the apoptosis against the Burkitt's lymphoma cells through the interaction of Gb3 express in their cell surface. On the other hand, SeviL obtained from gill and mantle of Mytilisepta virgata showed the specific binding against GM1b, asialo GM1 and SSEA-4 which are known as glycosphingolipid glycan including the β-galactoside. In addition, SeviL was identified as R type lectin by confirmation of QXW motif within its primary structure. Messenger RNA of SeviL like R type lectins was also found among the musssels including Mytilus galloprovincialis. SeviL also showed the apoptosis against asialo GM1 expressing cells. To apply the anticancer lectin as a novel molecular target drug, primary structure of MytiLec-1 was analyzed to enhance the stabilization of confirmation by computational design technique. It was succeeded to produce a monomeric artificial β-trefoil lectin, Mitsuba-1 without losing the Gb3 binding ability. Comparison of biological function between Mitsuba-1 and MytiLec-1 is also described in this study.

Effect of Autophagy to Drug Resistance of B-cell Lymphoma Cells

To investigate the effect of autophagy on the drug resistance of different human lymphoma cells. Human Burkitt's lymphoma cell Daudi, human B lymphoma cell SUDHL-4, and human mantle cell lymphoma cell JeKo-1 were taken as the research subjects. The expression of Atg5 was inhibited by the treatments of autophagy inhibitors or stable interference via lentivirus infection. The autophagy activity of B lymphoma cell was changed, and the changes of lymphoma cells to the drug resistance of ADR and VCR was observed. JeKo-1 cells showed the strongest resistance to ADR and VCR, followed by SUDHL-4, and Daudi cells showed the weakest resistance to ADR and VCR. At the same time, JeKo-1 cells showed the strongest autophagy activity, followed by SUDHL-4, and Daudi cells showed the weakest autophagy activity. After the treatments of autophagy inhibitors or stable Atg5 interference, the resistance of lymphoma cells to ADR and VCR was significantly weakened, and there was the positive correlation at the drug resistance and the autophagy activity of B lymphoma cell. The higher autophagy activity in lymphoma cells, the lower chemotherapy resistance of the lymphoma cells after autophagy was inhibited.

Synthesis of a reusable composite of graphene and silver nanoparticles for catalytic reduction of 4-nitrophenol and performance as anti-colorectal carcinoma

The preparation and formulation of new chemotherapeutic drugs and supplements with remarkable effects for the treatment of cancer are in the priority of both developing and developed countries. In the recent study, a composite of graphene and silver nanoparticles (GO/AgNPs) was fabricated to treat colorectal carcinoma. Characterization of GO/AgNPs was done by Field Emission Scanning Electron Microscopes (FE-SEM), Energy-Dispersive X-ray Spectroscopy (EDS), Transmission Electron Microscopy (TEM), X-Ray Diffraction (XRD), and Ultraviolet–Visible Spectrophotometry (UV–Vis) analysis. The catalytic activity of the GO/AgNPs nanocomposite is demonstrated in reducing 4-nitrophenol (4-NP) by NaBH4 as the effective reductant. The reaction was thoroughly studied over a UV–Vis spectrophotometer. The rate constants were also determined following pseudo-first-order kinetics and the catalyst was reused for several times with almost reproducible yields indicating the robustness of our catalyst. The anti-colorectal carcinoma potentials of GO, AgNO3, and GO/AgNPs were assessed by MTT assay against colorectal adenocarcinoma (HT-29), colorectal carcinoma (HCT 116), ileocecal colorectal adenocarcinoma (HCT-8 [HRT-18]), and Burkitt's lymphoma (Ramos.2G6.4C10) cell lines. GO/AgNPs had excellent cell death and anti-human colorectal cancer effects against colorectal carcinoma cell lines. The IC50 of GO/AgNPs were 489, 617, 537, and 496 μg/mL against HT-29, HCT 116, HCT-8 [HRT-18], and Ramos.2G6.4C10 cell lines, respectively. Among the above cell lines, the best finding of anti-colorectal cancer activities of GO/AgNPs was gained in the cell line of HT-29. The antioxidant activities of GO, AgNO3, and GO/AgNPs were investigated using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) test in comparison with butylated hydroxytoluene as a positive control. In the antioxidant test, GO/AgNPs prevented 50% of the DPPH molecules in the 344 μg/mL concentration. According to the above findings, it appears GO/AgNPs may be administrated as a chemotherapeutic supplement/drug to treat colorectal carcinoma.

Comparison of SYK Signaling Networks Reveals the Potential Molecular Determinants of Its Tumor-Promoting and Suppressing Functions.

Spleen tyrosine kinase (SYK) can behave as an oncogene or a tumor suppressor, depending on the cell and tissue type. As pharmacological SYK inhibitors are currently evaluated in clinical trials, it is important to gain more information on the molecular mechanisms underpinning these opposite roles. To this aim, we reconstructed and compared its signaling networks using phosphoproteomic data from breast cancer and Burkitt lymphoma cell lines where SYK behaves as a tumor suppressor and promoter. Bioinformatic analyses allowed for unveiling the main differences in signaling pathways, network topology and signal propagation from SYK to its potential effectors. In breast cancer cells, the SYK target-enriched signaling pathways included intercellular adhesion and Hippo signaling components that are often linked to tumor suppression. In Burkitt lymphoma cells, the SYK target-enriched signaling pathways included molecules that could play a role in SYK pro-oncogenic function in B-cell lymphomas. Several protein interactions were profoundly rewired in the breast cancer network compared with the Burkitt lymphoma network. These data demonstrate that proteomic profiling combined with mathematical network modeling allows untangling complex pathway interplays and revealing difficult to discern interactions among the SYK pathways that positively and negatively affect tumor formation and progression.

Anti-human colon cancer properties of a novel chemotherapeutic supplement formulated by gold nanoparticles containing Allium sativum L. leaf aqueous extract and investigation of its cytotoxicity and antioxidant activities

Recently, scientists have tried to increase organic chemistry productions for the treatment of many cancers such as colon cancers. Au nanoparticles have a special role in the treatment of several cancers. Furthermore, one of the therapeutic properties of Allium sativum L. is increasing the physiological potentials of the body against different cancers. In the current research, gold nanoparticles were prepared and synthesized in aqueous medium using Allium sativum L. leaf extract. Also, we investigated the anti-colon cancers potentials of these nanoparticles against colon cancer cell lines. The FT-IR, UV–Vis, FE-SEM, and TEM tests used to determine the physiochemical properties of the recent nanoparticles. The results of FE-SEM and TEM images indicated the average size of 19 nm. DPPH test was carried out to assess the antioxidant capacities of HAuCl4, A. sativum, and AuNPs. DPPH test revealed similar antioxidant potentials for A. sativum, AuNPs and butylated hydroxytoluene. For the determining of anti-colon cancer properties of HAuCl4, A. sativum, and AuNPs, MTT assay was used on HUVECs (Normal), HT-29 (Colorectal adenocarcinoma), HCT 116 (Colorectal carcinoma), HCT-8 [HRT-18] (Ileocecal colorectal adenocarcinoma), and Ramos.2G6.4C10 (Burkitt's lymphoma) cell lines. AuNPs had very low cell viability and anti-colon cancer effects dose-dependently against HT-29, HCT 116, HCT-8 [HRT-18], and Ramos.2G6.4C10 cell lines. The best result of anti-colon cancer activities of AuNPs against above cell lines was observed in the case of the HCT 116 cell line. In conclusion, the synthesized AuNPs revealed remarkable anti-colon cancer properties against colon cell lines. It seems the above nanocomposite can be used as a novel chemotherapeutic material in the future.

Nanoencapsulated rituximab mediates superior cellular immunity against metastatic B-cell lymphoma in a complement competent humanized mouse model

BackgroundDespite the numerous applications of monoclonal antibodies (mAbs) in cancer therapeutics, animal models available to test the therapeutic efficacy of new mAbs are limited. NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) mice are one...

The Role of P53 Serine 46, Burkitt Cell Lymphoma 2 (BCL-2), and Cysteinyl Aspartate Specific Proteinase 3 (Caspase 3) Proteins as Risk Factors in Abortion

An Incomplete abortion is one of the complications that often occur during pregnancy, but the biomolecularmechanism is still unknown. It has proven to be an increase in apoptosis activity that is controlled by avariety of proteins, including p53 Ser 46, Bcl-2, and caspase 3, in uteroplasenta and reproductive organs. Theaim of the study is to measure the level of protein p54 Ser 46, Bcl-2, and caspase 3 in abortion and normalpregnancy. A case control study conducted in 38 cases of incomplete abortion and 38 normal pregnancies asa control. The level of protein of Ser 46, Bcl-2, and caspase 3 examined by ELISA independent/Duplo. Thehigher level of p53 Ser 46 (&gt; 0,09 U/mL) led to higher risks of abortion with OR 8.52; 95% CI, 2.65 to 28.6.The higher level of Bcl-2 led to higher risks of abortion with OR 3.75; 95% CI, 1.24 to 11.68. The higherlevel of caspase 3 (&gt; 7 mg/mL) and p53 Ser 46 (&gt; 0,09 U/mL) led to higher risks of abortion with OR 71.15;95% CI, 8.74 to 578.92; with the result of series p, p, 0.001; p &lt; 0.008; and p = 0.001. Level of protein p53Ser 46 and caspase 3 in abortion is higher than normal pregnancies, while the Bcl-2 protein levels in abortionis lower than normal pregnancies.

Triclosan induces apoptosis in Burkitt lymphoma-derived BJAB cells through caspase and JNK/MAPK pathways.

Burkitt's lymphoma (BL) is the fastest growing human tumor. Current treatment consists of a multiagent regimen of cytotoxic drugs with serious side effjects including tumor lysis, cardiotoxicity, hepatic impairment, neuropathy, myelosuppression, increased susceptibility to malignancy, and death. Furthermore, therapeutic interventions in areas of BL prevalence are not as feasible as in high-income countries. Therefore, there exists an urgent need to identify new therapies with a safer profile and improved accessibility. Triclosan (TCS), an antimicrobial used in personal care products and surgical scrubs, has gained considerable interest as an antitumor agent due to its interference with fatty acid synthesis. Here, we investigate the antitumor properties and associated molecular mechanisms of TCS in BL-derived BJAB cells. Dose-dependent cell death was observed following treatment with 10-100µM TCS for 24h, which was associated with membrane phospholipid scrambling, compromised permeability, and cell shrinkage. TCS-induced cell death was accompanied by elevated intracellular calcium, perturbed redox balance, chromatin condensation, and DNA fragmentation. TCS upregulated Bad expression and downregulated that of Bcl2. Moreover, caspase and JNK MAPK signaling were required for the full apoptotic activity of TCS. In conclusion, this report identifies TCS as an antitumor agent and provides new insights into the molecular mechanisms governing TCS-induced apoptosis in BL cells.

Transcription factor 3 (TCF3) combined with histone deacetylase 3 (HDAC3) down-regulates microRNA-101 to promote Burkitt lymphoma cell proliferation and inhibit apoptosis

ABSTRACT To explore the function of transcription factor 3 (TCF3) on the proliferation and apoptosis of Burkitt lymphoma cells and its mechanism. qRT-PCR was performed to determine the expression of TCF3, histone deacetylase 3 (HDAC3), and microRNA-101 (miR-101) in the Burkitt lymphoma (BL) tumor tissues and lymph node tissues with reactive lymph node hyperplasia (RLNH). We found that the expression of TCF3 and HDAC3 was up-regulated in BL tumor tissues and lymphoma cells, and the miR-101 expression was down-regulated. And TCF3 and HDAC3 were negatively correlated with the expression of miR-101, respectively. In addition, knockdown of TCF3 can inhibit BL cell proliferation, reduce cell viability and promote cell apoptosis, retain the cell cycle in the G0/G1 phase, and inhibit the expression of Akt/mTOR pathway-related proteins (p-Akt and p-mTOR). When miR-101 was overexpressed, the results were the same as when TCF3 was knocked down. Moreover, we used Co-immunoprecipitation (Co-IP) to detect the interaction between TCF3 and HDAC3, and performed the Chromatin immunoprecipitation (ChIP) experiment to detect the enrichment of TCF3 and HDAC3 in the promoter region of miR-101. We found that TCF3 can interact with HDAC3 and is enriched in the miR-101 promoter region. In conclusion, TCF3 combined with HDAC3 down-regulates the expression of miR-101, thereby promoting the proliferation of BL cells and inhibiting their apoptosis.

Nicotinamide adenine dinucleotide phosphate oxidase inhibitor induces apoptosis on Epstein-Barr virus positive B lymphoma cells.

Over-expression of nicotinamide adenine dinucleotide phosphate oxidase (Nox) isoform enzymes was recently reported in various cancers including Burkitt’s lymphoma (BL). However, the functions of Nox isoform enzymes in BL remain poorly understood. In this study, Nox isoform expression and the effects of a Nox-specific inhibitor were evaluated in Epstein-Barr virus (EBV)-positive Raji BL cells in comparison with EBV-negative Ramos BL cells. To evaluate Nox enzyme expression in Raji and Ramos BL cells, polymerase chain reaction (PCR) and western blot analysis were performed. To verify the intracellular signaling mechanism of the Nox inhibitor-induced apoptosis of Raji cells, WST-1 assay, trypan blue exclusion method, flow cytometry, PCR, western blotting, and bromodeoxyuridine staining were conducted. Experiments using the pan-caspase inhibitor z-VAD, reactive oxygen species scavenger N-acetyl-L-cysteine (NAC), and Bim inhibitor 1 were performed. PCR and western blot results showed that Nox isoform enzymes were highly expressed in EBV-positive BL Raji cells compared with EBV-negative BL Ramos cells. The Nox2 inhibitor induced apoptosis of Raji cells in time- and dose-dependent manners. The Nox2 inhibitor also caused up-regulation of Bim and Noxa, down-regulation of Mcl-1, translocation of Bax, release of cytochrome c, and caspase cascade activation, resulting in apoptosis. Furthermore, z-VAD, NAC, and BI-1 effectively blocked the Nox2 inhibitor-induced apoptosis of Raji cells. Taken together, these results provide a novel insight into the mechanism of Nox inhibitor-induced apoptosis and evidence for Nox as a therapeutic target to treat EBV-positive malignancies.

Expression pattern of MRPS18 family genes in malignantly transformed b-cells.

To compare expression patterns of proteins of a family of mitochondrial ribosomal protein S18 (MRPS18) in tumor cell lines of the B-cell origin. The study has been performed on different subsets of tonsil B-cells and tumor cell lines of the B-cell origin using quantitative polymerase chain reaction analysis, western blot analysis, immunohistochemistry, bioinformatic analysis of the publicly available data bases on expression. We have found that genes of the MRPS18family (1-3) show different expression patterns in tumor cell lines of the B-cell origin. The highest levels of expression were shown for MRPS18-3, the lowest- for MRPS18-1. MRPS18-2was expressed at the highest levels in germinal center cells, Burkitt lymphoma and Hodgkin lymphoma cell lines. At the protein levels, MRPS18-2showed the highest expression in Burkitt lymphoma and B-cell precursor acute lymphoblastic leukemia cell lines. In lymphoblastoid cell lines, and in germinal center B-cells MRPS18-2levels were somewhat lower, but higher than in memory and plasma B-cells. The differential expression pattern of the MRPS18family proteins suggests that they play various roles in cellular processes.

Knockdown of long non‑coding RNA ANRIL inhibits the proliferation and promotes the apoptosis of Burkitt lymphoma cells through the TGF‑β1 signaling pathway.

Burkitt lymphoma (BL) has a high mortality rate and its treatment is currently limited to chemotherapy combined with immunotherapy. The long non-coding RNA antisense non-coding RNA in the INK4 locus (ANRIL) has been identified as an oncogene that can regulate cell proliferation and apoptosis in multiple types of cancer. However, the function of ANRIL in BL remains unknown. The present study aimed to determine the effect of ANRIL on cell proliferation and apoptosis in BL. Reverse transcription-quantitative PCR was used to analyze the expression levels of ANRIL in BL cells. The effect of ANRIL knockdown on BL cells was determined using Cell Counting Kit-8, flow cytometric, western blotting, immunofluorescence staining and Hoechst staining assays. The results revealed that ANRIL silencing inhibited the proliferation and promoted the apoptosis of BL cells. In addition, the expression levels of cyclin D1, E2F transcription factor 1 and Bcl-2 were downregulated, while the expression levels of cyclin-dependent kinase inhibitor 1A, Bcl-2-associated X protein, cleaved-caspase-9/pro-caspase-9 and cleaved-caspase-3/pro-caspase-3 were upregulated. Furthermore, the knockdown of ANRIL activated the TGF-β1 signaling pathway, as evidenced by the upregulated expression levels of TGF-β1, phosphorylated (p)-SMAD2/3/SMAD2/3, p-SMAD1/SMAD1 and sphingosine-1-phosphate receptor 2. Moreover, the protective effect of ANRIL silencing in BL could be inhibited by the TGF-β receptor type I/II dual inhibitor, LY2109761. In conclusion, the findings of the present study suggested that the knockdown of ANRIL may inhibit cell proliferation and promote cell apoptosis in BL by regulating the TGF-β1 signaling pathway, which may provide a novel target for the treatment of BL.