Leukemia K562 Research Articles

Exploring the anticancer potential of extracts and compounds from the heartwood of Cotinus coggygria Scop. wild growing in Serbia.

Cotinus coggygria has a long history of use in traditional medicine in Europe and Asia. The aim of study was to explore the cytotoxicity of extracts (EE-ethanol, MME-methylene chloride/methanol, and WE-water) and compounds (butin, butein, fisetin, sulfuretin, taxifolin, eriodictyol, fustin, cotinignan A, sulfuretin auronol, 3-O-methylepifustin, 3-O-methylfustin, and sitosterol-3-O-β-D-glucoside) isolated from C. coggygria. Mechanisms of anticancer effects of three extracts, butin, butein, and sulfuretin were examined. Compounds were isolated from the EE using silica gel column chromatography and semipreparative HPLC. Structure elucidation was performed using NMR spectroscopy. Cytotoxicity was evaluated using MTT assay. The effects on cell cycle and cell death were investigated by flow cytometry. The antimigration effects were examined by scratch assay, while expression of the MMP2, MMP9, and VEGFA were measured by quantitative real time PCR. The antioxidant effects were examined by flow cytometry. 3-O-methylepifustin, epitaxifolin, and sulfuretin auronol were found for the first time in C. coggygria. The extracts and compounds showed selective cytotoxicity against HeLa, MDA-MB-231, HL-60, K562, A375, PC-3, and DU 145 cells. HeLa cells were the most sensitive to the cytotoxicity of MME (IC50 value of 47.45µg/mL), while leukemia K562 and HL-60 cells were the most sensitive to the MME and EE (IC50 values in the range from 31.04 to 44.57µg/mL). Butein exerted strong cytotoxicity on HeLa, K562, and MDA-MB-231 cells (IC50 values of 8.66 µM, 13.91 µM, and 22.36 µM). EE, butin, butein, sulfuretin, and fisetin were highly selective against leukemia K562 cells when compared with normal fibroblasts MRC-5 (selectivity index: 4.01, 5.15, 6.17, 7.05, > 4.41, respectively). Butein and fisetin showed high selectivity in the cytotoxic activity against HeLa cells when compared with MRC-5 cells (selectivity index: 9.91 and > 6.61). Three extracts, butin, butein, and sulfuretin, initiated apoptosis in HeLa cells by activating caspase-8 and caspase-9. The extracts, butin, butein, and sulfuretin inhibited HeLa cell migration. EE, MME, butein, and sulfuretin exerted cytoprotective effects in normal fibroblasts. This research might suggest promising anticancer effects and underscores the need for additional research on C. coggygria extracts and compounds to assess their potential in cancer prevention and therapy.

Proliferation Inhibited by Genipin in Human Leukemia K562 Cells: Involvement of Uncoupling Protein 2 in Mitochondrial Damage.

Uncoupling protein 2 (UCP2) is essential for maintaining redox homeostasis and regulating energy metabolism. Abnormal expression of UCP2 has been associated with various tumors, including leukemia. Genipin (GEN), a specific inhibitor of UCP2, has a long history of use in traditional Chinese medicine. However, the precise role and underlying mechanisms of UCP2 in the inhibition of leukemia cells by GEN remain inadequately understood. This study focuses on the expression levels of UCP2 in myeloid leukemia (ML) and investigates the effects of GEN on the proliferation, mitochondrial function, and energy metabolism of the chronic myeloid leukemia (CML) cell line K562. The expression of UCP2 in clinical samples and cell lines (HL-60, U937, and K562) was confirmed using real-time quantitative polymerase chain reaction (qPCR) and western blot. The effects of GEN on K562 cell viability, morphology, and apoptosis were assessed through a cell counting kit-8 (CCK-8), Wright-Giemsa staining, and an annexin V-fluorescein isothiocyanate/propidium iodide (FITC/PI) apoptosis detection kit. Additionally, the impact of GEN on mitochondrial function and energy metabolism, including reactive oxygen species (ROS), mitochondrial membrane permeability transition pore (MPTP), lactic acid (LA), oxygen consumption rate (OCR), and adenosine triphosphate (ATP) levels in K562 cells, was also examined. The results showed that UCP2 was differentially expressed in clinical samples from patients with ML. Among the three cell lines examined, K562 cells exhibited a significantly higher expression level of UCP2. Functionally, GEN markedly inhibited K562 cell viability while promoting K562 cell differentiation and apoptosis. Mechanistically, UCP2 mRNA and protein expression levels were inhibited by GEN in K562 cells in a concentration- and time-dependent manner. Additionally, GEN dramatically increased ROS generation and induced mitochondrial MPTP opening in K562 cells. Furthermore, GEN significantly reduced LA production in K562 cells and markedly increased OCR and ATP production. The results suggest that UCP2 is differentially expressed in ML patients and cell lines; GEN, a UCP2 inhibitor, induces mitochondrial damage and metabolic remodeling, thereby inhibiting proliferation and promoting apoptosis in K562 cells, and thus could be suggested as an adjuvant of an antitumor metabolic therapy.

Total Synthesis of the Tridecasaccharide Motif fromAngelica sinensisAPS-1 II Polysaccharide with Anti-leukemia Activity and Structure-Activity Relationship Studies.

A polysaccharide APS-1 II from a medicinal plant Angelica sinensisrepresents an interesting therapeutic agent against leukemia. However, the synthetic accessibility of the highly branched and complex APS-1 II polysaccharide with multiple 1, 2-cis-glycosidic linkages remains a difficult task, impeding the in-depth structure-activity relationship biological studies and the development of carbohydrates-based therapeutics against leukemia. Here, we report the first chemical synthesis of tridecasaccharide repeating unit together with shorter sequences 4-mer, 6-mer and 9-mer from APS-1 II polysaccharide via one-pot orthogonal glycosylation strategy based on glycosyl ortho-(1-phenylvinyl)benzoates, which precluded the potential issues such as aglycone transfer associated with one-pot assembly with thioglycosides. The synthetic pathway also features the following aspects: 1) three contiguous and challenging 1, 2-cis-Fuc bonds were highly stereoselectively constructed via the newly developed stereoselective 1, 2-cis-fucosylation method; 2) several 1, 2-trans-glycosidic linkages were formed via neighboring group participation effect, while 1,2-cis-Glc linkage was stereoselectively assembled via N,N-dimethylformamide reagent modulation; 3) the final [1+1+2+9] one-pot assembly of the target tridecasaccharide via strategic utilizations of glycosyl N-phenyltrifluoroacetimidates, ortho-alkynylbenzoates and ortho-(1-phenylvinyl)benzoates. Biological studies revealed that human leukemia K562 and mouse L1210 cells could be effectively inhibited by tridecasaccharide repeating unit andnonasaccharide.

L-Asparaginase Immobilized on Nanographene Oxide as an Efficient Nanobiocatalytic Tool for Asparagine Depletion in Leukemia Cells.

l-Asparaginase (l-ASNase) catalyzes the hydrolysis of l-asparagine, leading to its depletion and subsequent effects on the cellular proliferation and survival. In contrast to normal cells, malignant cells that lack asparagine synthase are extremely susceptible to asparagine deficiency. l-ASNase has been successfully employed in treating pediatric leukemias and non-Hodgkin lymphomas; however, its usage in adult patients and other types of cancer is limited due to significant side effects and drug resistance. Recent research has explored alternative formulations and delivery methods to enhance its efficacy and minimize adverse effects. One promising approach involves the immobilization of l-ASNase onto nanostructured materials, offering improved enzymatic activity and biocompatibility of the support. We harnessed an E. coli l-ASNase type II preparation to develop a novel strategy of enzyme immobilization on graphene oxide (GO)-based support. We compared GO and nanographene oxide (nGO) in terms of their biocompatibility and influence on enzyme parameters. The obtained l-ASNase on the nGO nanobiocatalyst maintains enzymatic activity and increases its stability, selectively acting on K562 leukemia cells without cytotoxic influence on normal endothelial cells. In the case of treated K562 cells, we confirmed enlargement in the cell and nucleus size, disturbance in the cell cycle (interphase and metaphase), and increased apoptosis rate. The potential therapeutic possibilities of immobilized l-ASNase on leukemia cell damage are also discussed, highlighting the importance of further research in this area for advancing cancer therapy.

Quinizarin Gold(I) N-Heterocyclic Carbene Complexes with Synergistic Activity Against Anthracycline-Resistant Leukaemia Cells: Synthesis and Biological Activity Studies.

New, asymmetric quinizarin-Au(I)-NHC complexes were designed, isolated, and fully characterised including by single crystal X-ray crystallography. Cytotoxicity studies showed effective growth inhibition in HeLa cervical cancer cells with IC50 values ranging from 2.4 μM to 5.3 μM. The successful cellular uptake was evidenced by X-ray fluorescence imaging on cryo-preserved whole HeLa cells and the sub-cellular localisation was monitored by live-cell fluorescence microscopy. Notably, complex 2 b showed circumvention of acquired anthracycline resistance in K562 leukaemia cells as well as synergistic activity with doxorubicin against both wild-type and anthracycline-resistant Nalm-6 leukaemia cells. Interestingly, sub-cellular localisation towards mitochondria proved to be more important than the compounds' overall cytotoxicity for potent antiproliferative activity and to achieve effective resistance circumvention.

Oxabicyclo Isolate from the Leaves of Siparuna cymosa

In this research, the phytochemical study of extracts from the leaves of Siparuna cymosa, an endemic species in Brazil, was carried out for the first time, and evaluated the cytotoxic activity of this extract against leukemia cell lines. The dried leaves were powdered and subjected to extraction by exhaustive maceration with pure organic solvents. The chloroform and ethyl acetate extracts were purified by column chromatography, and the isolated compounds were identified by spectroscopic techniques. The cytotoxic activity of extracts and compounds against THP-1 (acute myeloid) and K-562 (chronic myeloid) leukemia lines was determined using 3-(4,5-dimethyl-2-thyazolyl)- 2,5-diphenyl-tetrazoliumbromide (MTT) assay. The isolated compounds were 1,5,6-trimethyl8-oxabicyclo[3.2.1]oct-6-en-3-ol, β-sitosterol-3-O-β-D-glucopyranoside, and kaempferol-3,7-diO-α-L-rhamnoside. This is the first report of isolation of oxabicyclo-octenol in plant species. β-Sitosterol-3-O-β-D-glucopyranoside and kaempferol-3,7-di-O-α-L-rhamnoside significantly reduced the viability of tumor cells after controlled exposure for 48 h. Moreover, selectivity was at least two times higher for such lineages compared to the results found for non-tumor peripheral blood cells (PBMC). Kaempferol-3,7-di-O-α-L-rhamnoside was the most active and selective. In conclusion, metabolites from S. cymosa showed to be of scientific interest. This study shows that the biodiversity of plant species in Brazil represents an important arsenal of organic compounds and, therefore, research should be motivated aiming at the isolation of secondary metabolites.

Flow cytometry conjugate formation assay between natural killer cells and their target cells.

Before being able to kill other cells, natural killer (NK) cells first have to establish contact with those targets. In case of a predominance of activating signals from the target cell over inhibitory ones, the killing process is initiated. It is possible, with a simple two-color flow cytometry method, to evaluate, for any given effector cell-target cell pair, the number of conjugates between both types of cells. The percentage obtained gives an idea of the amplitude of binding of the NK cells to the targets and might be expected to be indicative of the level of cytotoxicity. Nevertheless, there is no absolute correlation, as the percentages of conjugates are sometimes higher with relatively resistant targets than with the highly sensitive cell line K562. Practically, NK cells and target cells are stained with two differently fluorescent dyes and incubated together at the desired effector:target ratio (in our example, 1:1) for various periods of time (0, 10, 30min, etc.) at 37°C. After the incubation time, the cells are carefully introduced into the flow cytometer, where in principle three populations are distinguished: the single positive, unconjugated effector and target cells, respectively, and the double positive subset, which corresponds to the conjugates between both cell types. We describe here in detail the staining and cell culture protocols and procedures, and give several examples. Thus, the very cytotoxic NK leukemia cell line KHYG-1 versus the myeloid leukemia K562 (the "conventional" NK cell target) and the Burkitt lymphoma cell line Raji forms a high number of conjugates. In contrast, purified, non-activated, healthy donor-derived peripheral blood NK cells bind less to the targets, in accordance with their low (K562) or absent (Raji) cytotoxic activity.

Quantitative elemental analysis of human leukemia K562 single cells by inductively coupled plasma mass spectrometry in combination with a microdroplet generator

A μDG was installed into a sample introduction system of scICP-MS to achieve efficient and quantitative elemental analysis of cultured mammalian cells at the single-cell level.

Analysis of synthetic polymer hydrogel-based generation of leukemia stem cells.

Leukemia stem cells (LSCs), capable of simultaneous self-renewal and differentiation, are resistant to chemotherapy and the cause of relapse in refractory cases of leukemia. As a method to rapidly generate LSCs has not been established, research on LSCs as therapeutic targets has been hampered. Here, we demonstrate that K562 leukemia cells acquired LSC properties with increase in stemness markers such as CD34, Oct3/4, and Nanog and metabolic alterations towards OXPHOS by culturing cells on synthetic polymer hydrogels. In this hydrogel-generated LSCs, single-cell RNA sequencing identified the increase in expression levels of AKR1B1 and TSPYL5, which play an essential role for stemness generation. Decrease in expression of CD34, Oct3/4, and Nanog were observed in K562cells with knockdown of AKR1B1 and TSPYL5. These results indicate that cell culturing on synthetic polymer hydrogels can be a useful system to generate LSCs and AKR1B1 and TSPYL5 may become therapeutic targets for LSCs.

Synthesis and Anti-Cancer Activity In Vitro of Synephrine Derivatives.

Glucocorticoids (GCs) are routinely used to treat hematological malignancies; however, long-term treatment with GCs can lead to atrophic and metabolic adverse effects. Selective glucocorticoid receptor agonists (SEGRAs) with reduced side effects may act as a superior alternative to GCs. More than 30 SEGRAs have been described so far, yet none of them reached clinical trials for anti-cancer treatment. In the present work, we propose a novel approach to increase the number of potential SEGRAs by obtaining derivatives of synephrine, a molecule of natural origin. We synthesized 26 novel compounds from the class of synephrine derivatives and characterized them by HRMS, and 1H and 13C NMR. We evaluated in vitro anti-cancer effects in leukemia K562 and lymphoma Granta cells using the MTT assay and studied their potential affinity for the glucocorticoid receptor (GR) in silico using the molecular docking approach. The novel derivative 1-[4-(benzyloxy)phenyl]-2-(hexylamino)ethanol (10S-E2) with the highest GR affinity in silico exhibited cytotoxic activity against K562 and Granta cells after 24 h of treatment at the concentration of approximately 13 µM which correlated with its highest MolDock Score. The other compound with high GR affinity, 2-(hexylamino)-1-(4-nitrophenyl)ethanol (13S-G2), demonstrated cytotoxicity in both cell lines at concentrations of 50-70 µM. Overall, our results may provide a solid rationale for developing and further investigating synephrine derivatives as SEGRAs with anti-cancer activity.

Microfluidic impedance flow cytometer leveraging virtual constriction microchannel and its application in leukocyte differential

Microfluidic impedance flow cytometry has been widely used in leukocyte differential and counting, but it faces a bottleneck due to the trade-off between impedance detection throughput and sensitivity. In this study, a microfluidic impedance flow cytometer based on a virtual constriction microchannel was reported, in which the virtual constriction microchannel was constructed by crossflow of conductive sample and insulated sheath fluids with underneath micro-electrodes for impedance measurements. Compared to conventional mechanical constriction microchannels, this virtual counterpart could effectively avoid direct physical contact between cells and the microchannel walls to maintain high throughputs, and significantly reduce the volume of the impedance detection region for sensitivity improvements. Using the developed microfluidic impedance flow cytometer, impedance pulses of three leukemia cell lines, K562, Jurkat, and HL-60, were detected, achieving a 99.8% differentiation accuracy through the use of a recurrent neural network. Furthermore, impedance pulses of four white blood cell subpopulations (neutrophils, eosinophils, monocytes, and lymphocytes) from three donors were detected, achieving a classification accuracy of ≥99.2%. A classification network model was established based on purified white blood cell and applied to impedance pulses of two white blood cell mixtures, resulting in proportional distributions of four leukocyte subpopulations within theoretical ranges. These results indicated that the developed microfluidic impedance flow cytometer based on the virtual constriction microchannel could achieve both high detection throughput and high sensitivity, showing great potentials for clinical diagnostics and blood analysis.

Cellular uptake of CPX-351 by scavenger receptor class B type 1-mediated nonendocytic pathway

The proper uptake of drugs in liposome formulations into target cells markedly impacts therapeutic efficacy. The protein corona (PC), formed by the adsorption of serum proteins onto the liposome surface, binds to specific surface receptors of target cells, influencing the uptake pathway. We investigated the uptake pathway into leukemia cells based on PC analysis of CPX-351, a liposome containing cytarabine and daunorubicin in a fixed 5:1 synergistic molar ratio. The PC of CPX-351 mixed with fetal bovine serum was analyzed by nanoflow liquid chromatography-tandem mass spectrometry. CPX-351 uptake in HL-60, K562, and THP-1 leukemia cell lines, was measured by flow cytometry using daunorubicin fluorescence. The major components of CPX-351 PC were apolipoproteins A-I and A-II, which bind to scavenger receptor class B type 1 (SR-BI), a nonendocytic pathway that takes up only liposome contents. SR-BI was expressed in each cell, and its expression correlated with CPX-351 uptake. The uptake was significantly decreased by the inhibition of clathrin-mediated endocytosis and macropinocytosis. Additionally, blocks lipid transport-1 (BLT-1), a selective inhibitor of SR-BI, decreased the uptake; however, high-dose BLT-1 addition significantly increased the uptake, which was more strongly inhibited by macropinocytosis suppression compared with clathrin-mediated endocytosis. BLT-1 enhances the binding of SR-BI to liposomes in a dose-dependent manner. These findings indicate that the enhancement of binding between SR-BI and CPX-351 activates different pathways, such as macropinocytosis, distinct from CPX-351 alone. SR-BI may be a biomarker for CPX-351 therapy, and the combination of CPX-351 with high-dose BLT-1 may augment therapeutic efficacy.

Retraction Note: Silencing of membrane-associated sialidase Neu3 diminishes apoptosis resistance and triggers megakaryocytic differentiation of chronic myeloid leukemic cells K562 through the increase of ganglioside GM3

Retraction Note: Silencing of membrane-associated sialidase Neu3 diminishes apoptosis resistance and triggers megakaryocytic differentiation of chronic myeloid leukemic cells K562 through the increase of ganglioside GM3

Cancer-associated SF3B1 mutation K700E causes widespread changes in U2/branchpoint recognition without altering splicing.

Myelodysplastic syndromes and other cancers are often associated with mutations in the U2 snRNP protein SF3B1. Common SF3B1 mutations, including K700E, disrupt SF3B1 interaction with the protein SUGP1 and induce aberrant activation of cryptic 3' splice sites (ss), presumably resulting from aberrant U2/branch site (BS) recognition by the mutant spliceosome. Here, we apply the new method of U2 IP-seq to profile BS binding across the transcriptome of K562 leukemia cells carrying the SF3B1 K700E mutation. For cryptic 3' ss activated by K700E, we identify their associated BSs and show that they are indeed shifted from the WT sites. Unexpectedly, we also identify thousands of additional changes in BS binding in the mutant cells that do not alter 3' ss choice. These new BS are usually very close to the natural sites, occur upstream or downstream, and either exhibit stronger base-pairing potential with U2 snRNA or are adjacent to stronger polypyrimidine tracts than the WT sites. The widespread imprecision in BS recognition induced by K700E with limited changes in 3' ss selection supports a positive role for SUGP1 in early BS choice and expands the physiological consequences of this oncogenic mutation.

Targeting Poly(U) Binding Splicing Factor 60 (PUF60): A Small-Molecule Inhibitor Shows Anti-Leukemic Activity and Impacts Cell Cycle in Leukemia Models

The emerging role of altered RNA splicing in the development of myeloid neoplasms including MDS and AML has received wide attention. Facilitating genes, including the frequently mutated splicing factors, SF3B1, U2AF1, and SRSF2, have been reported to play a key role in leukemogenesis. Pharmacological intervention targeting cells harboring these mutations has been pursued, but limited agents have been reported. Rare events of more than one hit in splicing factors were found in MDS and AML patients' samples. This suggests that inhibiting another target in the splicing process may overcome the threshold of survival in these vulnerable cells carrying splicing factor mutations and induce toxicity. The concept has been demonstrated in the case of leukemia cells carrying U2AF1 mutations being vulnerable to inhibitors modulating wild-type SF3B1 function. The primary function of U2AF1 is to form a complex with U2AF2 for 3‘ splice site (SS) definition in RNA splicing and lower frequency U2AF2 mutations are also detected in MDS patients, underlying the significance of dysfunctional 3‘ SS definition as a potential facilitator of leukemogenesis. Here, we hypothesize that compounds inhibiting proteins participating in 3' SS selection may disrupt isoform patterns in leukemia cells. These agents may act alone or serve as a second hit in the RNA processing to invoke additional vulnerabilities to leukemia cells carrying the most frequent splicing factor mutations. Definition of 3' SS by U2AF2/U2AF1 is mediated by the recognition of the polypyridine tract (PPT) by U2AF2 and the binding of U2AF1 to the conserved AG dinucleotides at the 3' SS. Like U2AF2, PUF60 contains two zinc-finger domains, recognizing the polyuridine tract preceding 3' SS, and a U2 homology motif (UHM) domain that engages in protein-protein interaction with partner proteins including U2AF2, SF1, and SF3B1. Different from the U2AF1/U2AF2 complex, PUF60 does not have a protein domain to bind to the exon intron boundary junction at the 3' SS. PUF60 has been reported to enhance the splicing activity of U2AF1/U2AF2, but also directly regulates alternative splicing of a subset of genes. A recent finding indicated that knockout of PUF60 leads to alternative splicing switching of CDC25 from isoform A to isoform C to induce cell cycle arrest at G2/M in solid tumors. Leukemia patients carrying PUF60 overexpression have also been found to have less progression free survival (https://ualcan.path.uab.edu/cgi-bin/TCGA-survival1.pl?genenam=PUF60&ctype=LAML) compared to patients with low expression of PUF60. To support our hypothesis, we undertook an inhibitor development campaign to discover small-molecule compounds selectively targeting PUF60. Based on a relatively less selective and weak inhibitor to the UHM domain, SF-153, we conducted chemical modifications and obtained a PUF60 inhibitor, SF2-69, with >15-fold selectivity against U2AF1, RBM39, SPF45, and U2AF2. We found SF2-69 had an IC50 value of ~3 mM in NKM-1 and K562 leukemia cell lines. In the cell cycle analysis, we found SF2-69 increased S and subG1 cell population in NKM-1 at 24h, but induced G1 arrest in K562 at 48h in a dose dependent manner. In comparison, E7820, an RBM39 degrader, increased G2/M phase in both NKM-1 and K562. RNA-seq analysis of NKM-1 treated with SF2-69 after 24h revealed that SF2-69 upregulated a set of genes participating in cholesterol biosynthesis, NME1-NME2 (a tumor suppressor kinase) and downregulated PLK1, a kinase that phosphorylates and activates CDC25C in G2/M transition. Because K562 is p53-null and NKM-1 has wild-type p53, SF2-69 likely induced p53-dependent and p53-independent responses in NKM-1 and K562 cells, respectively. In summary, SF2-69 is a new selective PUF60 inhibitor which disrupts cell cycle progression in leukemia cell lines by modulating p53-mediated responses. Upregulation of cholesterol biosynthesis may be a feedback loop to counteract the inhibition of PUF60 by SF2-69. Further optimization of SF2-69 will allow us to develop more effective and selective chemical probes for studying the role of PUF60 overexpression in leukemia and the potential use of PUF60 inhibitors in MDS and AML cells carrying splicing factor mutations.

The combination of venetoclax and quercetin exerts a cytotoxic effect on acute myeloid leukemia

Venetoclax is a BH3 mimetic that was recently approved for the treatment of acute myeloid leukemia (AML) treatment. However, the effect of venetoclax on AML remains limited, and a novel strategy is required. Here, we demonstrate for the first time that the cytotoxic effect of venetoclax drastically increased when by combined with the naturally occurring flavonoid quercetin. Combined treatment with venetoclax and quercetin caused most of AML KG-1 cells to exhibit a condensed morphology. Cell cycle analysis revealed that the combination strongly induced cell death. Caspase inhibitor blocked this cell death, and the combination induced poly (ADP-ribose) polymerase (PARP) cleavage, indicating that apoptosis was the primary mechanism. These effects were also observed in another AML cell line Kasumi-1 but not in chronic myeloid leukemia (CML) K562 cells. Public data analysis demonstrated that B-cell/CLL lymphoma 2 (Bcl-2) expression is increased in AML cells compared to other malignant tumors, and the survival and the growth of AML cell line depends on Bcl-2. We found that quercetin increased Bcl-2-associated X protein (Bax) expression in KG-1. Our study provides a novel function for quercetin and presents a promising strategy for AML treatment using venetoclax.

One‐Pot Synthesis of [(l‐GluA)‐b‐(PCL)]‐Based Diblock Copolymer and Solvent‐Assisted Multifaceted Capsules as Potential Cargo Payload for Nanomedicines

Abstract This study shows that the synthesis of di‐BCP between amino acid l‐glutamic acid (l‐GluA) and hydrophobic polycaprolactone (PCL) leads to multifunctional and multifaceted capsule formation with different dispersion solvents. Successful synthesis of [(l‐GluA)‐b‐(PCL)] was confirmed by using FTIR and 1H NMR characterizations. For morphological studies, synthesized di‐BCP of [(l‐GluA)‐b‐(PCL)] was dispersed in four different solvents such as acetone, DMSO, IPA, and ethanol and was used for sample preparation for SEM imaging. Interestingly, smooth‐surfaced NPs (defined as type I NPs), rough‐surfaced NPs (type II NPs), porous NPs (type III NPs), and football‐type deep excavation having NPs (defined as type IV NPs). Furthermore, these NPs were characterized through BET, AFM, and DLS for particle size and ζ potential of NPs. LC for types III and IV NPs was calculated to be 1.96% ± 0.01% and 1.92% ± 0.01%, respectively. EE of types III and IV NPs was calculated to be 91% ± 1.5% and 94% ± 1.2%. We used K562 (leukemia blood cancer) and HEK293 (embryonic kidney) cells for in vitro studies. DOX@[(l‐GluA)‐b‐(PCL)] nanoformulation related to types III and IV NPs shows significant early apoptosis that is, 29.17%, which is a remarkable programmed cell death for these porous nanoparticles.

Chlorine containing tetrahydropyrimidines: Synthesis, characterization, anticancer activity and mechanism of action

The aim of the presented research was to explore anticancer potential of eleven newly synthesized tetrahydropyrimidine derivatives. The compounds were synthesized via Biginelli multicomponent one-pot reaction using different derivatives of vanillin, ethyl 4-chloroacetoacetate and (N-methyl)urea. The cytotoxic effects of the compounds were examined on three human malignant cell lines (HeLa, K562, and MCF7), and normal lung fibroblasts MRC-5. The mechanisms of anticancer activity were examined for two compounds 4a and 4b which showed the strongest and selective cytotoxicity against chronic myelogenous leukaemia K562 cells (IC50 = 1.76 ± 0.09, and 1.66 ± 0.05, respectively). The changes of matrix metalloproteinase 2 (MMP2), matrix metalloproteinase 9 (MMP9), and vascular endothelial growth factor A (VEGFA) were investigated in the K562 cell line, as well as oncomiRNA miR-10b, miR-23a described to have both features, depending on a specific type of malignancy, and miR-34a with mostly described as a tumour suppressor.

Study of TRAIL and SAHA Co-Treatment on Leukemia K562 Cell Line.

TRAIL (Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand) is an attractive agent being considered a potential cancer treatment. It attaches to its death receptors, leading many cancer cells to apoptosis. However, some malignancies indicate substantial resistance to TRAIL, challenging anticancer scientists. Herein, combination therapy with TRAIL plus SAHA (Suberoyl Anilide Hydroxamic Acid) was conducted to evaluate the capability of SAHA to overcome TRAIL resistance in the leukemia K562 cell line. First, the IC50 for SAHA was calculated (2 µM) at 12, 24, 48, and 72 h of treatment using MTT assay. Second, the K562 cells were treated with concentrations of 50 and 100 nM of TRAIL and 2 μM of SAHA separately and together for 24, 48, and 72 h and the survival of these cells was evaluated by Flowcytometry following the annexin-V and PI staining. To demonstrate the non-toxicity of the combined treatment for normal cells, the HEK-293 cell line was treated with the TRAIL 100 nM and SAHA 2 μM combined and separated at the same periods. In the end, by performing real-time PCR, the amount of candidate genes' expression implicated in TRAIL resistance, and the levels of BCR-ABL expression was measured. The drug dosages were not toxic to normal cells. SAHA plus TRAIL strongly triggered apoptosis in K562 cells after 24, 48, and 72 h of exposure. Furthermore, it was shown that DR4, DR5, and CHOP expressions were enhanced, and PI3K, Akt, ERK, STAT3, c-FLIPL, NF-κB, and BCR-ABL expressions were decreased by SAHA in K562 cells. Our study indicated that SAHA combined with TRAIL can increase the sensitivity of K562 leukemic cells to TRAIL by suppressing intracellular anti-apoptotic molecules and augmenting the expressions of DR4/DR5 and CHOP.

Differentiating leukemia subtypes based on metabolic signatures using hyperpolarized 13C NMR.

Leukemia is a group of blood cancers that are classified in four major classes. Within these four classes, many different subtypes exists with similar origin, genetic mutations, and level of maturity, which can make them difficult to distinguish. Despite their similarities, they might respond differently to treatment, and therefore distinguishing between them is of crucial importance. A deranged metabolic phenotype (Warburg effect) is often seen in cancer cells, leukemia cells included, and is increasingly a target for improved diagnosis and treatment. In this study, hyperpolarized 13C NMR spectroscopy was used to characterize the metabolic signatures of the six leukemia cell lines ML-1, CCRF-CEM, THP-1, MOLT-4, HL-60, and K562. This was done using [1-13C]pyruvate and [1-13C]alanine as bioprobes for downstream metabolite quantification and kinetic analysis on cultured cells with and without 2-deoxy-D-glucose treatment. The metabolic signatures of similar leukemia subtypes could be readily distinguished. This includes ML-1 and THP-1, which are of the similar M4 and M5 AML subtypes, CCRF-CEM and MOLT-4, which are of the similar T-ALL lineage at different maturation states, and HL-60 and K562, which are of the closely related M1 and M2 AML subtypes. The data presented here demonstrate the potential of hyperpolarized 13C NMR spectroscopy as a method to differentiate between leukemia subtypes of similar origin. Combining this method with bioreactor setups could potentially allow for better leukemia disease management as metabolic signatures could be acquired from a single biopsy through repeated experimentation and intervention.