Quercetin Promotes Cell Cycle Arrest and Apoptosis and Attenuates the Proliferation of Human Chronic Myeloid Leukemia Cell Line-K562 Through Interaction with HSPs (70 and 90), MAT2A and FOXM1

2019 ◽  
Vol 19 (12) ◽  
pp. 1523-1534 ◽  
Author(s):  
Ali Hassanzadeh ◽  
Elham Hosseinzadeh ◽  
Saleheh Rezapour ◽  
Ghasem Vahedi ◽  
Navideh Haghnavaz ◽  
...  
Keyword(s):  
Caspase 3 ◽  
Kinase Inhibitors ◽  
Philadelphia Chromosome ◽  
Leukemia Cell ◽  
Annexin V ◽  
Leukemic Cells ◽  
Leukemia Cell Line ◽  
Genes Expression ◽  
K 562 Cells ◽  
Induced Apoptosis

Background: Chronic Myeloid Leukaemia (CML) starts in certain blood-forming cells of the bone marrow when cells acquire Philadelphia chromosome. Nowadays, scientists attempt to find novel and safe therapeutic agents and approaches for CML therapy using Tyrosine Kinase Inhibitors (TKIs), CML conventional treatment agents, has some restrictions and also adverse effects. Recently, it has been proposed that phytochemicals, such as flavonoids due to their low side effects and notable safety have the potential to be used for CML therapy. Materials and Methods: K-562 cells were exposed with three concentrations of the querectin (10, 40 and 80µM) for 12, 24 and 48 hours. After that, these cells apoptosis rate was estimated using Annexin-V/PI staining and flowcytometry analysis, and their proliferation rate was evaluated using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT). Finally, the expression of the 70 and 90 kilodalton heat shock proteins (HSP70 and 90), methionine adenosyltransferase 2A (MAT2A), Forkhead box protein M1 (FOXM1), caspase-3 and -8, Bcl-X(L) and Bax involved in leukemic cells survival and proliferation was assessed using Real-Time PCR within 12, 24 and 48 hours after exposure with quercetin 40 and 80µM. Results: Considering consequences, querecetin induced apoptosis in K-562 cells, and also abrogated these cells proliferation. On the other hand, RT-PCR results showed a reduction in some of the candidate genes expression, especially HSP70, Bcl-X(L) and FOXM1, when cells were treated with quercetin 40 and 80µM. Also, Bax, caspase-3 and caspase-8 expression was significantly improved in K-562 cells upon quercetin exposure. Conclusion: We concluded that CML therapy by querecetin due to its anti-proliferative and anti-survival potentials could lead to the promising therapeutic outcome through targeting major survival and proliferation involved genes expression.

scholarly journals Digoxin-Like Immunoreactive Factors Induce Apoptosis in Human Acute T-Cell Lymphoblastic Leukemia

2007 ◽  
Vol 53 (7) ◽  
pp. 1315-1322 ◽  
Author(s):  
Kenneth Ihenetu ◽  
Hassan M Qazzaz ◽  
Fabian Crespo ◽  
Rafael Fernandez-Botran ◽  
Roland Valdes
Keyword(s):  
T Cell ◽  
Cell Line ◽  
Lymphoblastic Leukemia ◽  
Leukemia Cell ◽  
Jurkat Cells ◽  
Leukemic Cells ◽  
Leukemia Cell Line ◽  
Induction Of Apoptosis ◽  
K 562 Cells ◽  
Induced Apoptosis

Abstract Background: Plant-derived cardenolides reportedly possess anticancer properties in human leukemic cells via selective induction of apoptosis, cell cycle arrest, and differentiation. Selective induction of apoptosis with mammalian-derived digoxin-like immunoreactive factor (DLIF) could provide new strategies for anticancer drug development or the identification of biomarkers for cancer. We investigated whether DLIFs selectively induce apoptosis in human lymphoblastic leukemic cells. Methods: We compared the relative potencies of digoxin, ouabain, and DLIF on induction of programmed cell death in Jurkat cells (an acute T-leukemic cell line), K-562 (a myelogenous leukemia cell line), and nonpathologic human peripheral blood mononuclear cells (PBMCs). Apoptosis was measured by flow cytometry with the annexin V/propidium iodide method. Results: Digoxin and ouabain induced apoptosis in Jurkat cells [digoxin 50% inhibitory concentration (IC50), 24 nmol/L; ouabain IC50, 26 nmol/L]. Neither digoxin nor ouabain induced apoptosis in K-562 cells or PBMCs. DLIF was more potent (IC50, 1.9 nmol/L) and >2-fold more effective than digoxin or ouabain at inducing maximum apoptosis in Jurkat cells. The IC50 values in the apoptosis assays were >100-fold lower (DLIF) and 20-fold lower (digoxin and ouabain) than the IC50 required for Na+- and K+-dependent ATPase (DLIF, 200 nmol/L; digoxin, 910 nmol/L; ouabain, 600 nmol/L). Conclusion: DLIF selectively induces apoptosis in a human acute T-cell lymphoblastic leukemia cell line but not in K-562 cells or PBMCs. These data suggest a new physiological role for these endogenous hormone-like factors.

The SirT1 Inhibitor Tenovin-6 Induces Monocytic Differentiation in APL Cells

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 1542-1542
Author(s):  
Yoshitaka Sunami ◽  
Marito Araki ◽  
Soji Morishita ◽  
Yumi Hironaka ◽  
Yoko Edahiro ◽  
...  
Keyword(s):  
Conflicts Of Interest ◽  
Leukemia Cell ◽  
Annexin V ◽  
Leukemia Cell Line ◽  
Limited Information ◽  
Monocytic Differentiation ◽  
Nb4 Cells ◽  
Downstream Target ◽  
Altered Cell ◽  
Induced Apoptosis

Abstract Abstract 1542 Tenovin-6, an inhibitor for nicotinamide adenine dinucleotide (NAD)-dependent histone deacetylase Sirtuins, shows cytotoxicity to cancerous cells and thus recognized as a potential therapeutic compound for cancer treatment (Lain S., et al. (2008) Cancer Cell. 13,454–63). Since there is limited information for the cytotoxic property of this compound for hematopetic malignancies, we treated an APL (acute promyelocytic leukemia) cell line NB4 with Tenovin-6. As expected, Annexin V assay revealed that Tenovin-6 induced apoptosis in NB4 cells. However, to our surprise, at modest concentration, Tenovin did not induce cell death, rather inhibited NB4 cell proliferation and altered cell morphology. The fluorescence-activated cell sorting (FACS) analysis revealed that tenovin-6-treated NB4 cells are positive for CD11b and CD36 with decreased level of CD13 and CD33. Moreover, tenovin-6-treated NB4 cells presented nitroblue tetrazolium reduction capacity, suggesting that tenovin-6 induced monocytic differentiation in NB4 cells. To assess how Tenovin-6 induces cellular differentiation in NB4 cells, we investigated downstream target of Sirtuins. Although Tenovin-6 reportedly promotes acetylation of p53 by inhibiting SirT1, a founder of Sirtuin family proteins, the acetylation status of p53 is unchanged at the concentration where we observed differentiation. This suggests that Tenovin-6 induces NB4 differentiation through inhibiting other Sirtuin family proteins. These findings demonstrate a potential of Tenovin-6 as a differentiation-inducing reagent in APL cells. Disclosures: No relevant conflicts of interest to declare.

scholarly journals In Vitro Inhibitory Effect of Recombinant Human Calprotectin on Nalm6 Leukemia Cell Line

2020 ◽  
Vol 20 (8) ◽  
pp. 951-962
Author(s):  
Samira Charkhizadeh ◽  
Mehdi Imani ◽  
Nematollah Gheibi ◽  
Fateme Shabaani ◽  
Akbar Nikpajouh ◽  
...  
Keyword(s):  
Cell Line ◽  
Leukemia Cell ◽  
Annexin V ◽  
Inhibitory Effect ◽  
New Drugs ◽  
Leukemia Cell Line ◽  
Cytotoxic Effects ◽  
Tumor Expression ◽  
Induced Apoptosis

Background & Purpose: In evaluating new drugs for the treatment of various types of cancer, investigations have been made to discover a variety of anti-tumor compounds with less side effects on normal cells. Investigations have shown that the heterodimers S100A8 and S100A9 inhibit the enzyme casein kinase 2 and then prevent the activation of the E7 oncoprotein. Therefore, the aim of this study was to evaluate the effect of calprotectin as an antitumor compound on the Nalm6 (B cell precursor leukemia cell line). Material & Methods: Transformation of genes encoding S100A8 and S100A9 human, designed in the pQE32 plasmid, was performed by the thermal shock method into E. coli M15 bacteria. After bacterial growth in LB medium, the expression of two S100A8 and S100A9 subunits, the solubility of the protein by SDS-PAGE method was determined. Finally, the S100A8 / A9 complex was equally placed in the microtube. In the next step, the cytotoxic effects of calprotectin produced on the Nalm6 cell line were evaluated using the wst1 test. Then, the apoptosis in these cells was measured using flow cytometry methods with Annexin-V coloration. Results: In the current study, the results showed that the cytotoxic effects of Calprotectin are time and concentration- dependent. Therefore, it can reduce the tumor expression and had a beneficial effect by induced apoptosis in Nalm6 cell line. Conclusion: Calprotectin has an anti-tumor effect on the Nalm6 cell line by increasing apoptosis.

scholarly journals 3ʹS,4ʹS-Disenecioylkhellactone from the Roots of Peucedanum Japonicum Induces Apoptosis in HL-60 Cells ​

2020 ◽  
Author(s):  
Kyung-Yun Kang ◽  
Sung-Ju Lee ◽  
Hyun-Ji Kim ◽  
Heonjoong Kang ◽  
Sang-Jip Nam ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Caspase 3 ◽  
Biological Activities ◽  
Leukemia Cell ◽  
Annexin V ◽  
The Philippines ◽  
Cell Counting ◽  
Leukemia Cell Line ◽  
Resonance Spectroscopy ◽  
Human Leukemia

Abstract Background: The root of Peucedanum japonicum is used in traditional medicine in Japan, the Philippines, China, and Korea, and it has been reported to have a variety of biological activities, including anticancer activity. Inducing apoptosis in tumor cells has become a major focus of antitumor drug development; therefore, we studied the apoptotic effects of the MeOH/CH2Cl2 extract of the roots of P. japonicum and its components on HL-60 cells (a human leukemia cell line).Methods: Compounds were purified using solvents with varying polarities followed by column chromatography (reverse phase), and the structures were confirmed using nuclear magnetic resonance spectroscopy. The viabilities of HL-60, A549, and MCF-7 cells were determined using the cell counting kit-8 (CCK-8) assay. Analysis of apoptosis signaling was performed only with HL-60 cells, and cell cycle progression, Annexin V/propidium iodide (PI) staining (analyzed by flow cytometry), the mitochondrial membrane potential (MMP, as analyzed by flow cytometry), and caspase-3, 8, and 9 activity (determined using the caspase-3, -8, and -9 activity kit according to the manufacturer’s protocol) were evaluated.Results: Two coumarin molecules, (-)-isosamidin (1) and 3ʹS,4ʹS disenecioylkhellactone (2), were isolated by bioactivity-guided fractionation. Only compound 2 showed cytotoxicity in HL-60 cells, which occurred due to increases in the sub-G1 population and the initiation of early and late apoptosis as determined by Annexin V/PI staining. In addition, decreases in the MMP were observed in HL-60 cells treated with compound 2. Several apoptotic features were observed, including increased cleavage of caspase-3, -8, and -9. Moreover, treatment with Z-DEVD-FMK (a caspase-3 inhibitor), Z-IETD-FMK (a caspase-8 inhibitor), or Z-LEHD-FMK (a caspase-9 inhibitor) significantly inhibited cell cytotoxicity.Conclusions: We provide evidence that compound 2 induces apoptosis in HL-60 cells.

Ectopic Expression of Mer on Jurkat Cells and Its Anti-Apoptosis Effect.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4098-4098
Author(s):  
Lei Fan ◽  
Liqian Xie ◽  
Fei Shen ◽  
Lan Dai ◽  
Jianyong Li ◽  
...  
Keyword(s):  
T Cells ◽  
Tyrosine Kinase ◽  
Caspase 3 ◽  
Ectopic Expression ◽  
Leukemia Cell ◽  
Annexin V ◽  
Jurkat Cells ◽  
Leukemia Cell Line ◽  
Serum Starvation ◽  
Significant Difference

Abstract BACKGROUND & OBJECTIVE: Mer is one member of Axl receptor tyrosine kinase family; its ligand Gas6 can bind Mer, then stimulate the tyrosine kinase activity and downstream cell signal pathway of Mer, and take part in cell inflammation, apoptosis and thrombosis. There are more and more reports on Mer function, while few on its association with malignant diseases. This study was to detect the expression of Mer on T-cell leukemia cell line--Jurkat, and investigate its anti-apoptosis function and the mechanism. METHODS: Flow cytometry (FCM) was used to detect Mer expression on normal T cells and Jurkat cells. RNA interference (RNAi) was used to block the expression of Mer on Jurkat cells. The effect of Mer on the proliferation of Jurkat cells was assessed by MTT assay, and its effect on serum starvation-induced apoptosis was evaluated by FCM with Annexin V/PI double staining. The expression of apoptosis-associated genes Bcl-2 and Caspase-3 in Jurkat cells was detected by real-time polymerase chain reaction (PCR) after Mer blocking. RUSELTS: Mer was not expressed in normal T cells both in peripheral blood and bone marrow, but highly expressed in Jurkat cells with a positive rate of 51.1%±5.2%. The inhibition rate of Mer expression in Jurkat cells by RNAi was 86.0%±10.3%. After 48-hour serum starvation, the apoptosis rate was 15.3%±4.3% in Mer-blocking Jurkat cells, and only 1.5%±1.0% in control Jurkat cells(P<0.01). There was no significant difference in Jurkat cell proliferation rate between these 2 groups. After Mer blocking, Bcl-2 expression was decreased by 42.7%±8.6% of control(P<0.01), Caspase-3 expression showed no significant change. CONCLUSION: Mer is highly expressed in Jurkat cells, and could inhibit cell apoptosis via Bcl-2 signaling pathway.

Involvement of caspase-3 activation in squamocin-induced apoptosis in leukemia cell line HL-60

Life Sciences ◽  
2002 ◽  
Vol 70 (11) ◽  
pp. 1259-1269 ◽  
Author(s):  
Xiao-Feng Zhu ◽  
Zong-Chao Liu ◽  
Bin-Fen Xie ◽  
Zhi-Ming Li ◽  
Gong-Kan Feng ◽  
...  
Keyword(s):  
Cell Line ◽  
Caspase 3 ◽  
Leukemia Cell ◽  
Leukemia Cell Line ◽  
Induced Apoptosis

FLIPLong(L) and FLIPShort(S) Overexpression in the Human Myeloid Leukemia Cell Line ML-1 and Its Role in TRAIL [Tumor Necrosis Factor(TNF)-Related Apoptosis-Inducing Ligand] and TNFa Induced Apoptosis.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4378-4378
Author(s):  
Sudeshna Seal ◽  
Daniella B. Kerbauy ◽  
Vladimir Lesnikov ◽  
Nissa Abbasi-Shafer ◽  
H. Joachim Deeg
Keyword(s):  
Caspase 3 ◽  
Leukemia Cell ◽  
Leukemia Cell Line ◽  
Caspase 8 ◽  
Slight Reduction ◽  
Western Blots ◽  
Apoptosis Pathway ◽  
Induced Apoptosis ◽  
Active Caspase

Abstract TRAIL initiates activation of Caspase-8, which is blocked by the FLICE inhibitory protein (FLIP), resulting in resistance to apoptosis. Here we show that overexpression of FLIPL and FLIPS in ML1 cells, with low constitutive levels of FLIP, protects these cells against apoptosis induced by TRAIL, not only via Caspase-8 inhibition, but also via upregulation of anti-apoptotic molecules. Methods: 1) Apoptosis was determined by Annexin V/ 7-AAD following treatment with TRAIL (100–500ng/ml), or TNFa (20–100ng/ml) in ML1 cells transduced with FLIPL.GFP (green fluorescent protein), FLIPS.GFP or Neo.GFP (control). 2) Caspase-8, Caspase-3, Bid, Bcl-xL, XIAP, phosphorylated (P)-IKBa and P-Akt were determined by western blots. 3) Active Caspase-3 was determined using EnzChek Caspase-3 assay kit. Results: Both FLIPL and FLIPS transduction protected ML1 cells against apoptosis induced by TRAIL (300ng/ml), while no protection was observed in Neo.GFP cells. FLIPL had a more profound protective effect than FLIPS (Fig.1A). Both FLIPL and FLIPS, but not Neo.GFP, blocked Caspase-8 and Caspase-3 activation (Fig.1B); FLIPS cells showed two-fold higher levels of active Caspase-3 than FLIPL cells, consistent with higher apoptosis in FLIPS cells. Caspase-3 can be activated through Caspase-8 (extrinsic pathway) or via Caspase-8/Bid (intrinsic pathway). The latter was responsible for high active Caspase-3 levels in FLIPS cells as shown by the presence of cleaved Bid (t-Bid) (Fig.1B); cleavage of Bid was inhibited by combination of TRAIL and Z-IETD-FMK (Caspase-8 inhibitor). Anti-apoptotic molecules, including Bcl-xL, XIAP and FLIP are regulated by NF-kB and FLIP participates in an NF-kB auto-amplification loop. While Neo.GFP cells showed little Bcl-xL after 4h of TRAIL exposure and there was a twofold reduction in FLIPS cells, only a slight reduction of Bcl-xL was noted in FLIPL cells. FLIPL cells showed the lowest rates of apoptosis when exposed to TNFa and BMS543541, a specific inhibitor of IkB kinase (Fig. 1C). In the presence of BMS543541, phosphorylation of IkBa and levels of Bcl-xL and XIAP decreased in Neo.GFP and FLIPS but not in FLIPL cells. Additional data suggest that the PI3-kinase/Akt pathway is involved in constitutive NF-kB activation and differentially affected by FLIPL and FLIPS (Fig. 1D). Preliminary results in immunodeficient mice transplanted with transduced ML1 cells indicated the in vivo relevance of the differences between FLIPL and FLIPS with FLIPL cells engrafting earlier and showing earlier signs of sickness. Conclusions: FLIPL and FLIPS conferred resistance to TRAIL induced apoptosis but showed differential effects: Caspase-8/Bid was involved in the apoptosis pathway in FLIPS, but not in FLIPL cells. FLIPL cells’ resistance was due not only to caspase inhibition but to the recruitment of downstream anti-apoptotic pathways such as NF-kB and PI3K/Akt. In vivo data further substantiated the antiapoptotic/pro-survival behavior of FLIPL cells. Figure Figure

scholarly journals Human acute leukemia cell line with the t(4;11) chromosomal rearrangement exhibits B lineage and monocytic characteristics

Blood ◽  
1985 ◽  
Vol 65 (1) ◽  
pp. 21-31 ◽  
Author(s):  
RC Stong ◽  
SJ Korsmeyer ◽  
JL Parkin ◽  
DC Arthur ◽  
JH Kersey
Keyword(s):  
Acute Leukemia ◽  
Cell Line ◽  
Lymphoblastic Leukemia ◽  
Leukemia Cell ◽  
Leukemic Cells ◽  
Leukemia Cell Line ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Immunoglobulin Gene ◽  
A Cell ◽  
B Lineage

Abstract A cell line, designated RS4;11, was established from the bone marrow of a patient in relapse with an acute leukemia that was characterized by the t(4;11) chromosomal abnormality. The cell line and the patient's fresh leukemic cells both had the t(4;11)(q21;q23) and an isochromosome for the long arm of No. 7. Morphologically, all cells were lymphoid in appearance. Ultrastructurally and cytochemically, approximately 30% of the cells possessed myeloid features. The cells were strongly positive for terminal deoxynucleotidyl transferase. They were HLA-DR positive and expressed surface antigens characteristic for B lineage cells, including those detected by anti-B4, BA-1, BA-2, and PI153/3. Immunoglobulin gene analysis revealed rearrangements of the heavy chain and kappa chain genes. The cells lacked the common acute lymphoblastic leukemia antigen and antigenic markers characteristic of T lineage cells. The cells reacted with the myeloid antibody 1G10 but not with other myeloid monoclonal antibodies. Treatment with 12-O-tetradecanoyl- phorbol-13-acetate induced a monocyte-like phenotype demonstrated by cytochemical, functional, immunologic, and electron microscopic studies. The expression of markers of both early lymphoid and early myeloid cells represents an unusual phenotype and suggests that RS4;11 represents a cell with dual lineage capabilities. To our knowledge, RS4;11 is the first cell line established from t(4;11)-associated acute leukemia.

scholarly journals Human Platelets Effectively Kill K-562 Cells, a Chronic Myelogenic Leukemia Cell Line,in vitro

1990 ◽  
Vol 81 (5) ◽  
pp. 449-453 ◽  
Author(s):  
Terutaka Sagawa ◽  
Takeshi Kodama ◽  
Akio Tominaga ◽  
Mariko Okada
Keyword(s):  
Cell Line ◽  
Leukemia Cell ◽  
Human Platelets ◽  
Leukemia Cell Line ◽  
K 562 Cells
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