Functional Analysis of a Cation Transporter OCT6: A Mediator of the Cellular Influx of Adriamycin.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4374-4374
Author(s):  
Yoko Okitsu ◽  
Hideo Harigae ◽  
Shinichiro Takahashi ◽  
Shori Abe ◽  
Mitsunori Okabe ◽  
...  
Keyword(s):  
Functional Analysis ◽  
Xenopus Oocytes ◽  
Intracellular Concentration ◽  
Myelogenous Leukemia ◽  
Leukemic Cells ◽  
Leukemia Cell Line ◽  
Clinical Samples ◽  
Dependent Manner ◽  
Cancer Drugs ◽  
Anti Cancer

Abstract (Introduction) It is necessary for killing leukemic cells effectively to achieve a sufficient intracellular concentration of anti-cancer drugs. To date, overexpression of efflux pump such as MDR-1 and MRP-1, which leads to a decrease of cellular concentration of drugs, has been shown to attenuate the therapeutic effect of anti-cancer drugs and correlate with low remission rate in acute leukemia. If the expression of genes, which import anti-cancer drugs, can be induced specifically in leukemic cells, it would lead to an increase of intracellular concentration, resulting in an enhancement of the pharmacological effect. In this study, we performed the pharmacological characterization of a cation transporter gene, OCT6, and explore the possibility of its application for anti-cancer therapy. (Methods) Tissue distribution of OCT6 was examined by Northern blot analysis. Expression in clinical samples that consist of 34 patients; 15 patients with acute lymphoblastic leukemia (ALL), 17 patients with acute myelogenous leukemia (AML), 2 patients with chronic myelogenous leukemia (CML) in blastic crisis, was examined by quantitative RT-PCR. The percentage of blasts in each sample was above 80%. For functional analysis, OCT6 cRNA was injected into Xenopus oocytes and pharmacological analysis was performed. In order to examine the transporting activity in hematopoietic cells, we established subclones of Jurkat, a T lymphocytic leukemia cell line, in which OCT6 gene is constitutively expressed, and the change of the sensitivity for adriamycin was examined. (Result) By Northernblot analysis, 2.4kb OCT6 mRNA was detected in testis and bone marrow in adult tissues. OCT6 expressed in Xenopus oocytes transported not only tetraethlammonium but also adriamycin in a saturable and dose-dependent manner. The apparent Km value for [14C] adriamycin was 5.2±0.4μM. On the other hand, OCT6 did not transport either of methotrexate (MTX) and 5-fluorouracil (5-FU), which are not organic cations. After the exposure of adrimycin at the therapeutic concentration, the viability of OCT6 overexpressing Jurkat cells was significantly decreased compared to that of control cells. Consistent with this, the percentage of apoptosis cells became higher than that of control cells. When the expression level was examined in clinical samples, the average of the levels of patients was comparable to PBMCs. The level of patients, who achieved with complete remission (CR), was slightly higher than that of patients with non-response (NR), but the difference was not significant. Some patients exhibited the high expression of OCT6, but the distribution did not deviated to either CR or NR group. (Conclusion) These results suggest that the induction of OCT6 in leukemic cells might be a novel strategy for leukemia treatment.

scholarly journals Induction of apoptosis by Kola nut extract as a recent and promising treatment strategy for Leukemia

Bionatura ◽  
2021 ◽  
Vol 6 (2) ◽  
pp. 1725-1732
Author(s):  
Hamdah Alsaeedi ◽  
Rowaid Qahwaji ◽  
Talal Qadah
Keyword(s):  
Cell Cycle ◽  
Cell Line ◽  
Leukemia Cell ◽  
K562 Cells ◽  
Myelogenous Leukemia ◽  
Time Dependent ◽  
Leukemia Cell Line ◽  
Apoptotic Cells ◽  
Dependent Manner ◽  
Anti Cancer

Kola nut extracts have recently been reported to contain chemopreventive compounds providing several pharmacological benefits. This study investigated Kola nut extracts' anti-cancer activity on human immortalized myelogenous leukemia cell line K562 through apoptosis and cell cycle arrest. Fresh Kola nuts were prepared as powder and dissolved in DMSO. Different concentrations (50, 100, 150, 200, and 250 μg/ml) of working solutions were prepared. The K562 cells were treated with the different concentrations of Kola nut extract or vehicle control (10% DMSO) followed by incubation at 37°C for 24, 48, and 72 hours, respectively. Treatment activity was investigated in K562 cells; by Resazurin, and FITC/Propidium Iodide and 7-AAD stained cells to evaluate apoptotic cells and the cell cycle's progression. Inhibition of leukemia cell proliferation was observed. The extract effectively induced cell death, early and late apoptosis by approximately 30% after 24 and 48 hours incubation, and an increase in the rate of dead cells by 50% was observed after 72 hours of incubation. Also, cell growth reduction was seen at high dose concentrations (150 and 200 µg/ml), as evident by cell count once treated with Kola nut extract. The total number of apoptotic cells increased from 5.8% of the control group to 27.4% at 250 µg/ml concentration. Moreover, Kola nut extracts' effects on K562 cells increased gradually in a dose and time-dependent manner. It was observed that Kola nut extracts could arrest the cell cycle in the G2/M phase as an increase in the number of cells by 29.8% and 14.6 % were observed from 9.8% and 5.2% after 24 and 48 hours of incubation, respectively. This increase was detected in a dose and time-dependent manner. Kola nut extracts can be used as a novel anti-cancer agent in Leukemia treatment as it has shown significant therapeutic potential and therefore provides new insights in understanding the mechanisms of its action. Keywords: Kola nut extracts, Leukemia, K562 cell line, Apoptosis, Cancer.

scholarly journals Voruciclib, a Potent CDK4/6 Inhibitor, Antagonizes ABCB1 and ABCG2-Mediated Multi-Drug Resistance in Cancer Cells

2018 ◽  
Vol 45 (4) ◽  
pp. 1515-1528 ◽  
Author(s):  
Pranav Gupta ◽  
Yun-Kai Zhang ◽  
Xiao-Yu Zhang ◽  
Yi-Jun Wang ◽  
Kimberly W. Lu ◽  
...  
Keyword(s):  
Atpase Activity ◽  
Cellular Localization ◽  
Intracellular Localization ◽  
Scintillation Counter ◽  
Flow Cytometric Analysis ◽  
Multi Drug Resistance ◽  
Dependent Manner ◽  
Cancer Drugs ◽  
Intracellular Accumulation ◽  
Anti Cancer

Background/Aims: The overexpression of ATP-Binding Cassette (ABC) transporters has known to be one of the major obstacles impeding the success of chemotherapy in drug resistant cancers. In this study, we evaluated voruciclib, a CDK 4/6 inhibitor, for its chemo-sensitizing activity in ABCB1- and ABCG2- overexpressing cells. Methods: Cytotoxicity and reversal effect of voruciclib was determined by MTT assay. The intracellular accumulation and efflux of ABCB1 and ABCG2 substrates were measured by scintillation counter. The effects on expression and intracellular localization of ABCB1 and ABCG2 proteins were determined by Western blotting and immunofluorescence, respectively. Vanadate-sensitive ATPase assay was done to determine the effect of voruciclib on the ATPase activity of ABCB1 and ABCG2. Flow cytometric analysis was done to determine the effect of voruciclib on apoptosis of ABCB1 and ABCG2-overexpressing cells and docking analysis was done to determine the interaction of voruciclib with ABCB1 and ACBG2 protein. Results: Voruciclib significantly potentiated the effect of paclitaxel and doxorubicin in ABCB1-overexpressing cells, as well as mitoxantrone and SN-38 in ABCG2-overexpressing cells. Voruciclib moderately sensitized ABCC10- overexpressing cells to paclitaxel, whereas it did not alter the cytotoxicity of substrates of ABCC1. Furthermore, voruciclib increased the intracellular accumulation and decreased the efflux of substrate anti-cancer drugs from ABCB1- or ABCG2-overexpressing cells. However, voruciclib did not alter the expression or the sub-cellular localization of ABCB1 or ABCG2. Voruciclib stimulated the ATPase activity of both ABCB1 and ABCG2 in a concentration-dependent manner. Lastly, voruciclib exhibited a drug-induced apoptotic effect in ABCB1- or ABCG2- overexpressing cells. Conclusion: Voruciclib is currently a phase I clinical trial drug. Our findings strongly support its potential use in combination with conventional anti-cancer drugs for cancer chemotherapy.

scholarly journals Autocrine stimulation of interleukin 1 beta in acute myelogenous leukemia cells.

1987 ◽  
Vol 166 (5) ◽  
pp. 1597-1602 ◽  
Author(s):  
K Sakai ◽  
T Hattori ◽  
M Matsuoka ◽  
N Asou ◽  
S Yamamoto ◽  
...  
Keyword(s):  
Acute Myelogenous Leukemia ◽  
Interleukin 1 ◽  
Myelogenous Leukemia ◽  
Leukemic Cells ◽  
Dependent Manner ◽  
Acute Myelogenous ◽  
Autocrine Stimulation ◽  
Basal Proliferation ◽  
Culture Supernatants ◽  
Dose Dependent

A significant increase in CD25 antigen-positive cells by IL-1 was observed in cells of a patient with M7 acute myelogenous leukemia. Basal proliferation and expression of CD25 antigen by the M7 leukemic cells were inhibited by addition of anti-IL-1 beta antibody in a dose-dependent manner, but not by rabbit anti-IL-1 alpha antibody. Culture supernatants of these leukemic cells contained IL-1 activity, which was specifically inhibited by addition of anti-IL-1 beta antibody, and Northern blot analysis detected intracellular IL-1 beta mRNA. These results indicated that autocrine secretion of IL-1 beta was involved in proliferation of some myelogenous leukemic cells.

Identification of the Epigenetic Reader BRD4 As a Novel Potential Target in Ph+ CML

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 1571-1571
Author(s):  
Barbara Peter ◽  
Gregor Eisenwort ◽  
Gabriele Stefanzl ◽  
Daniela Berger ◽  
Wolfgang R Sperr ◽  
...  
Keyword(s):  
Cell Lines ◽  
Research Funding ◽  
Drug Target ◽  
K562 Cells ◽  
Myelogenous Leukemia ◽  
Leukemic Cells ◽  
Thymidine Uptake ◽  
Dependent Manner ◽  
Potential Drug ◽  
Ku812 Cells

Abstract Chronic myelogenous leukemia (CML) is a bone marrow-derived hematopoietic neoplasm in which BCR/ABL1 acts as a major driver of proliferation, differentiation and survival of leukemic cells. In a majority of all patients with CML, leukemic cells can be kept under control by BCR/ABL1 tyrosine kinase inhibitors (TKI), including imatinib, nilotinib, dasatinib, bosutinib, and ponatinib. Nevertheless, resistance or intolerance against one or more of these TKI may occur. Therefore, current research is focusing on novel potential drug targets in CML. A promising class of targets may be epigenetic regulators of cell growth, such as members of the bromodomain and extra-terminal domain (BET) family. The epigenetic reader and BET family member BRD4 has recently been identified as a novel potential drug target in acute myeloid leukemia (AML). However, so far, little is known about the expression and function of BRD4 in CML cells. The aims of the present study were to determine the expression of BRD4 and its downstream target MYC in CML cells and to explore whether BRD4 can serve as a novel drug target in this disease. As determined by qPCR, primary CML cells (chronic phase patients, n=7) as well as the CML cell lines KU812 and K562 expressed BRD4 mRNA. In addition, both CML cell lines stained positive for BRD4 in our immunocytochemistry staining experiments. In one patient with accelerated phase CML, putative leukemic (CD34+/CD38-) stem cells were sorted to near homogeneity and found to express BRD4 mRNA by qPCR. In order to examine the functional role of BRD4 in CML cells, a BRD4-specific shRNA was applied. In these experiments, the shRNA-induced knockdown of BRD4 in KU812 cells and K562 resulted in reduced growth compared to a control shRNA. Furthermore, the BRD4-targeting drug JQ1 was found to inhibit 3H-thymidine uptake and thus proliferation in KU812 cells in a dose-dependent manner (IC50: 0.25-0.75 µM). In addition, we were able to show that JQ1 inhibits growth of primary CML cells with variable IC50 values (0.1-5 µM). However, no substantial growth-inhibitory effects of JQ1 were seen in K562 cells (IC50: >5 µM). As determined by Annexin V/PI staining, JQ1 induced apoptosis in KU812 cells whereas no apoptosis-inducing effect of JQ1 was observed in K562 cells. Nevertheless, we were able to show that both CML cell lines as well as primary CML cells express MYC mRNA, and treatment of KU812 cells or K562 cells with JQ1 resulted in a decreased expression of MYC mRNA and MYC protein. Next, we analyzed whether MYC expression in CML cells can be blocked by BCR/ABL1 TKI. We found that imatinib, nilotinib, dasatinib, and ponatinib decrease MYC mRNA- and MYC protein expression in KU812 and K562 cells. Finally, we found that JQ1 cooperates with imatinib, nilotinib, ponatinib and dasatinib in inhibiting the proliferation of KU812 and K562 cells. Together, our data show that BRD4 serves as a potential new target in CML cells, and that the BRD4 blocker JQ1 cooperates with BCR/ABL1 TKI in inducing growth-inhibition. Whether BRD4 inhibition is a pharmacologically meaningful approach in patients with TKI-resistant CML remains to be determined in clinical trials. Disclosures Sperr: Ariad: Consultancy; Celgene: Consultancy. Zuber:Mirimus Inc.: Consultancy, Other: Stock holder; Boehringer Ingelheim: Research Funding. Valent:Novartis: Consultancy, Honoraria, Research Funding; Ariad: Honoraria, Research Funding; Bristol-Myers Squibb: Honoraria; Pfizer: Honoraria; Celgene: Honoraria.

Intracellular concentrations of anti cancer drugs in leukemic cells in vitro vs in vivo

1990 ◽  
Vol 25 (4) ◽  
pp. 252-256 ◽  
Author(s):  
B. Sundman-Engberg ◽  
U. Tidefelt ◽  
J. Liliemark ◽  
C. Paul
Keyword(s):  
Leukemic Cells ◽  
Cancer Drugs ◽  
Anti Cancer ◽  
Intracellular Concentrations

scholarly journals An undifferentiated variant derived from the human acute myelogenous leukemia cell line (KG-1)

Blood ◽  
1980 ◽  
Vol 56 (2) ◽  
pp. 265-273 ◽  
Author(s):  
HP Koeffler ◽  
R Billing ◽  
AJ Lusis ◽  
R Sparkes ◽  
DW Golde
Keyword(s):  
Cell Line ◽  
Acute Myelogenous Leukemia ◽  
Cellular Response ◽  
Leukemia Cell ◽  
Hla Antigens ◽  
Myelogenous Leukemia ◽  
Leukemic Cells ◽  
Leukemia Cell Line ◽  
Acute Myelogenous

Abstract A variant subline (KG-1a) of the human acute myelogenous leukemia (AML) cell line (KG-1) has been isolated. The cells retain the same constitutive markers as the parent line, including HLA antigens, isoenzymes, and karyotype. The cells from the subline are morphologically and histochemically undifferentiated blast cells, while the parent cells and several of its clones are at the myeloblast and promyelocyte stages of development. The variant cells do not respond to colony-stimulating factor (CSF), and they do not express the human la antigen, nor a recently characterized AML antigen. The parent KG-1 cells are stimulated to proliferate in the presence of CSF and the cells express the la and AML antigen. Variant AML cell lines, such as KG-1a, will be useful in vitro models for investigating cellular response to CSF and for studying antigen expression in leukemic cells.

Combined therapy of ruthenium dendrimers and anti-cancer drugs against human leukemic cells

2021 ◽  
Author(s):  
Sylwia Michlewska ◽  
Marta Maroto ◽  
Marcin Hołota ◽  
Małgorzata Kubczak ◽  
Natalia Sanz del Olmo ◽  
...  
Keyword(s):  
Combined Therapy ◽  
Leukemic Cells ◽  
Cancer Drugs ◽  
Anti Cancer ◽  
Human Leukemic Cells

Carbosilane ruthenium(II) dendrimers have been complexed with conventional anti-cancer drugs. Due to its features, the presence of ruthenium within a dendrimer structure improves the anti-cancer properties of nanocomplexes containing 5-flurouracyl,...

Mechanism Study of HL-60 Leukemic Cells Proliferative Inhibition Induced by Chinese Medicinal Herbs Mixture Jieduweikang Pill.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4512-4512
Author(s):  
Ri Zhang ◽  
Pin Wu ◽  
Zi-ling Zhu ◽  
Jiannong Cen ◽  
De Pei Wu
Keyword(s):  
Colony Formation ◽  
Herbal Remedy ◽  
Leukemia Cell ◽  
Leukemic Cells ◽  
Leukemia Cell Line ◽  
Dependent Manner ◽  
Dna Ladder ◽  
Antitumor Effects ◽  
Concentration Dependent Manner ◽  
Nbt Reduction

Abstract Background. It’s not surprising that the traditional Chinese herbal remedy is yielding useful drug leads. The mixture composed of thirteen Chinese medicinal plants---jieduweikang(JDWK) pill including Qingdai, Qinghao, Huangchi, Baihuasheshecao, Banzhilian and so on has been shown the clinical antitumor effects in China. We here report the mechanism investigation of JDWK pill cytotoxicity to leukemia cell line HL-60. Methods By using the method of serum pharmacology, the serum containing the JDWK pill was collected from rats. The MTT and tumor colony formation were used to determine the effects of JDWK pill on proliferation of HL-60 cells. NBT reduction, DNA electrophoresis, flow cytometry and RT-PCR were used to analyze the differentiation and apoptosis of HL-60 cells after treating with JDWK pill. Elisa was used to assay the secretion of VEGF from HL-60 cells. Results The liquid growth and colony formation of HL-60 cells were evidently inhibited by the serum containing JDWK pill and the cells were shown characteristic DNA “ladder” on agarose gel electrophoresis and more than 20% apoptotic cells in DNA histogram as well as the downregulation of bcl-2 gene related to apoptosis in a concentration-dependent manner, compared with the control. NBT reduction assay indicated that JDWK pill with low concentration may induce myeloid terminal differentiation of HL-60 cells and the level of VEGF was obviously decresed. Conclusion The JDWK pill can exerts the its anti-proliferative effect on the leukemic HL-60 cells through inducing apoptosis relating to the downregulation of Bcl-2 gene and anti-angiogenic effects with higher concentration and inducing myeloid differentiation with lower concentration.

scholarly journals The product of the cbl oncogene forms stable complexes in vivo with endogenous Crk in a tyrosine phosphorylation-dependent manner.

1996 ◽  
Vol 16 (1) ◽  
pp. 45-52 ◽  
Author(s):  
V Ribon ◽  
S Hubbell ◽  
R Herrera ◽  
A R Saltiel
Keyword(s):  
Tyrosine Phosphorylation ◽  
Leukemia Cell ◽  
Physiological Role ◽  
Sh2 Domain ◽  
Myelogenous Leukemia ◽  
Leukemia Cell Line ◽  
Stable Complex ◽  
Binding Motif ◽  
Dependent Manner

The cellular homologs of the v-Crk oncogene product are composed exclusively of Src homology region 2 (SH2) and SH3 domains. v-Crk overexpression in fibroblasts causes cell transformation and elevated tyrosine phosphorylation of specific cellular proteins. Among these proteins is a 130-kDa protein, identified as p130cas, that forms a stable complex in vivo with v-Crk. We have explored the role of endogenous Crk proteins in Bcr-Abl-transformed cells. In the K562 human chronic myelogenous leukemia cell line, p130cas is not tyrosine phosphorylated or bound to Crk. Instead, Crk proteins predominantly associate with the tyrosine-phosphorylated proto-oncogene product of Cbl. In vitro analysis showed that this interaction is mediated by the SH2 domain of Crk and can be inhibited with a phosphopeptide containing the Crk-SH2 binding motif. In NIH 3T3 cells transformed by Bcr-Abl, c-Cbl becomes strongly tyrosine phosphorylated and associates with c-Crk. The complex between c-Crk and c-Cbl is also seen upon T-cell receptor cross-linking or with the transforming, tyrosine-phosphorylated c-Cbl. These results indicate that Crk binds to c-Cbl in a tyrosine phosphorylation-dependent manner, suggesting a physiological role for the Crk-c-Cbl complex in Bcr-Abl tyrosine phosphorylation-mediated transformation.

scholarly journals Anti-Cancer Effects of Protein Extracts fromCalvatia lilacina,Pleurotus ostreatusandVolvariella volvacea

2011 ◽  
Vol 2011 ◽  
pp. 1-10 ◽  
Author(s):  
Jin-Yi Wu ◽  
Chi-Hung Chen ◽  
Wen-Huei Chang ◽  
King-Thom Chung ◽  
Yi-Wen Liu ◽  
...  
Keyword(s):  
Cell Line ◽  
Colorectal Adenocarcinoma ◽  
Leukemia Cell Line ◽  
Dependent Manner ◽  
Ros Production ◽  
Monocytic Leukemia ◽  
Anti Cancer ◽  
Sw480 Cells ◽  
Po Protein ◽  
Human Monocytic Leukemia

Calvatia lilacina(CL),Pleurotus ostreatus(PO) andVolvariella volvacea(VV) are widely distributed worldwide and commonly eaten as mushrooms. In this study, cell viabilities were evaluated for a human colorectal adenocarcinoma cell line (SW480 cells) and a human monocytic leukemia cell line (THP-1 cells). Apoptotic mechanisms induced by the protein extracts of PO and VV were evaluated for SW480 cells. The viabilities of THP-1 and SW480 cells decreased in a concentration-dependent manner after 24 h of treatment with the protein extracts of CL, PO or VV. Apoptosis analysis revealed that the percentage of SW480 cells in the SubG1phase (a marker of apoptosis) was increased upon PO and VV protein-extract treatments, indicating that oligonucleosomal DNA fragmentation existed concomitantly with cellular death. The PO and VV protein extracts induced reactive oxygen species (ROS) production, glutathione (GSH) depletion and mitochondrial transmembrane potential (ΔΨm) loss in SW480 cells. Pretreatment withN-acetylcysteine, GSH or cyclosporine A partially prevented the apoptosis induced by PO protein extracts, but not that induced by VV extracts, in SW480 cells. The protein extracts of CL, PO and VV exhibited therapeutic efficacy against human colorectal adenocarcinoma cells and human monocytic leukemia cells. The PO protein extracts induced apoptosis in SW480 cells partially through ROS production, GSH depletion and mitochondrial dysfunction. Therefore, the protein extracts of these mushrooms could be considered an important source of new anti-cancer drugs.

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