The aim of this study was to observe the inhibitory effect of daunorubicin on KG1a cells and the expression of Eps8 which is a novel tumor-associated antigen with its full name epidermal growth factor receptor pathway substrate 8 (Eps8), and to explore the effect of daunorubicin on Eps8 expression in KG1a cells at mRNA and protein levels. The KG1a cells were treated with different concentration of daunorubicin for 24, 48, 72 h, then trypan blue staining was used to detect the inhibitory rate of KGla cells, RQ-PCR and Western blot were used to detect Eps8 mRNA and Eps8 protein expression. The results showed that daunorubicin inhibited the proliferation of KG1a cells in a dose and time dependent manner (r = 0.983, P < 0.01). Daunorubicin could reduce the mRNA and protein levels of Eps8 expression in dose and time dependent manners in KG1a cells (r = 0.979, P < 0.05). It is concluded that with the increasing of concentration and time of daunorubicin acting on KG1a cells, the cell proliferative inhibitory effect increased and the expression of Eps8 decreased, suggesting that the inhibitory effect of daunorubicin on KG1a cell proliferation is realized through downregulation of Eps8 expression.
Adaptor Proteins, Signal Transducing ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Daunorubicin ; pharmacology ; Humans

The aim of this study was to investigate the effect of sorafenib combined with daunorubicin on leukemic k562 cell line. The inhibitory effect of sorafenib alone and its combination with daunorubicin on K562 cell proliferation was detected by MTT method; the synergistic effect was measured by CDI (coefficient of drug interaction); the apoptosis of K562 cells was observed by flow cytometry with Hoechst 33258 staining. The results showed that the sorafenib alone or its combination with daunorubicin could significantly inhibit K562 cell proliferation and the combination of both drugs displayed synergistic effect on K562 cells, meanwhile the apoptotic cells increased. It is concluded that the combination of sorafenib and daunorubicin has a obviously synergistic inhibitory effect on leukemic cell line K562.
Apoptosis ; drug effects ; Benzenesulfonates ; pharmacology ; Daunorubicin ; pharmacology ; Drug Synergism ; Humans ; K562 Cells ; Niacinamide ; analogs & derivatives ; Phenylurea Compounds ; Pyridines ; pharmacology

OBJECTIVE: To investigate the effect and mechanism of artesunate (ARTS) combined with cytarabine(Ara-C) and/or daunorubicin (DNR) on the proliferation and apoptosis of MV4-11 human mixed-lineage leukemia rearranged（MLL-r） acute myeloid leukemia (AML) cell line. METHODS: CCK-8 assay was used to detect the proliferation effect of individual or in combination of ARTS, DNR, Ara-C on MV4-11 cells. The IC₅₀ of ARTS, DNR and Ara-C was calculated separately. The cell apoptosis and expression of receptors DR4 and DR5 were detected by flow cytometry. Western blot was used to detect the expression of Caspase-3 and Caspase-9 in each groups. RESULTS: The inhibition effect of ARTS, Ara-C and DNR on the proliferation of MV4-11 were all dose-dependently (r=0.99, 0.90 and 0.97, respectively). The IC₅₀ of ARTS, Ara-C and DNR on MV4-11 for 48 hours were 0.31 μg/ml, 1.43 μmol/L and 22.47 nmol/L, respectively. At the dose of ARTS 0.3 μg/ml， Ara-C 1.0 μmol/L and DNR 15 nmol/L, the proliferation rate for 48 hours of the tri-combination treatment was significantly lower than that of the bi-combination treatment, while both were significantly lower than that of the individual treatment (all P＜0.05). In terms of bi-combination treatment, the cells proliferation rate for 48 hours of the ARTS+Ara-C group was significantly lower than that of the ARTS+DNR group, while both were significantly lower than that of the Ara-C+DNR group (all P＜0.05). The cooperativity index (CI) of bi- and tri-combination treatment were all less than 1. After 48 hours of drug action, the cell apoptosis rate of the ARTS+DNR+Ara-C group was significantly higher than that of the Ara-C+DNR group, while both were significantly higher than that of the ARTS+DNR group (all P＜0.05). Meanwhile, the was no statistical difference between the cells apoptotic rate of the ARTS+DNR+Ara-C group and the ARTS+Ara-C group (P>0.05). The expression of DR4 and DR5 also showed no difference between control group and drug group. Compared with the DNR+Ara-C group, the expressions of Caspase-3 were significantly down-regulated in both the ARTS+DNR+Ara-C group and the ARTS+Ara-C group (all P＜0.05). The down-regulation of Caspase-3 expression was the most significantly in the combination group of three drugs, while the Caspase-9 expressions in different groups showed no apparent change. CONCLUSION The in vitro study showed that tri-combination of ARTS+Ara-C+DNR and bi-combination of ARTS+Ara-C could inhibit the proliferation and promote apoptosis of MV4-11 cell line. The inhibition effect of these two combinations were significantly superior to that of the traditional Ara-C+DNR treatment. The mechanism underlying this finding may be identified by the down regulation of Caspase-3, while no altered expression was observed of Caspase-9, DR4 and DR5.
Humans ; Cytarabine/pharmacology* ; Daunorubicin/pharmacology* ; Caspase 3 ; Caspase 9 ; Artesunate/pharmacology* ; Leukemia, Myeloid, Acute ; Apoptosis ; Cell Line

OBJECTIVETo explore whether daunorubicin (DNR) combined with cytosine arabinoside (Ara-C) and DNR alone have similar effect on acute promyelocytic leukemia (APL) cell line NB4 and acute myeloblastic leukemia cell line HL-60 in vitro.

METHODSCell morphology, cells viability, and cell apoptosis (Annexin-V by flow cytometry assay) were analysed.

RESULTSAfter incubation with DNR plus Ara-C for 24 hours,NB4 cell viability [(36.75 +/- 3.82)%] (n = 6) and cell apoptosis rate [(21.24 +/- 5.82)%] (n = 3) did not change significantly compared to that treated with DNR alone for 24 hours [(35.73 + 6.28 )%, (22.55 +/- 3.26)%, respectively] (P > 0.05). However, HL-60 cell viability [(67.17 +/- 2.07)%] and cell apoptosis rate [(48.05 +/- 0.92)%] changed significantly in DNR plus Ara-C group compared with DNR alone [(63.31 +/- 1.80)% ,(41.51 +/- 0.89)%, respectively] (P < 0.01 and < 0.05, respectively).

CONCLUSIONDNR plus Ara-C and DNR alone have similar effect on NB4 cells, but have different effect on HL-60 cells.

Apoptosis ; drug effects ; Cytarabine ; pharmacology ; Daunorubicin ; pharmacology ; HL-60 Cells ; drug effects ; Humans ; Leukemia, Promyelocytic, Acute ; pathology

To explore the relationship of multidrug resistance formation in K562/A02 cells with the intracellular concentration of [Ca(2+)]i, the cytotoxicities of daunorubicin (DNR) were assayed by MTT method, the variations of [Ca(2+)]i of K562 cells and K562/A02 cells after treatment of Tet, DRL and DNR alone or in combination were detected by using Fura-2/AM. The results showed as follows: (1) The cytotoxicities of DNR to cell line K562/A02 were enhanced by 1 micro mol/L Tet or 5 micro mol/L DRL. Their IC(50) was (7.28 +/- 2.06) micro g/ml and (7.58 +/- 3.44) micro g/ml; multiple of their reversal effect was 2.94 and 2.82, but IC(50) of combined Tet and DRL was (1.66 +/- 0.41) micro g/ml. Its reverse effect distinctly increased by 12.9 times. (2) The [Ca(2+)]i in K562/A02 cells were higher than that in K562 cells. (3) One micro mol/L Tet and 5 micro mol/L DRL alone increased the [Ca(2+)]i in K562/A02 cells time-dependently and there was antagonism when both were used. It is concluded that high [Ca(2+)]i is supposed to be a reason of MDR in K562/A02 cells, the action of resistance modifying agents (RMA) in MDR reverse course, however, needs further research.
Alkaloids ; pharmacology ; Benzylisoquinolines ; pharmacology ; Calcium ; metabolism ; Daunorubicin ; pharmacology ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Humans ; K562 Cells ; Tamoxifen ; analogs & derivatives ; pharmacology

This study was purposed to investigate the reversal effect of glucosylceramide synthase (GCS) inhibitor D, L-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP) hydrochloride, on multidrug resistance in K562/A02 cells and its mechanism. The IC(50) (the half maximal inhibitory concentration) of PDMP was measured by MTT method. Cell apoptosis and intracellular daunorubicin (DNR) concentration were detected by flow cytometry. The expression of GCS and mdr1 genes were assayed by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot. The results showed that the IC(50) of DNR in K562 and K562/A02 cells were 0.23 +/- 0.02 and 7.15 +/- 0.24 microg/ml respectively. When the concentration of PDMP was equal to or less than 20 micromol/L ( < / = 20 micromol/L), the obviously inhibitory effect on proliferation of K562 and K562/A02 cells was not observed, but both 20 micromol/L and 10 micromol/L PDMP could enhance the sensitivity of K562/A02 cells to DNR (p < 0.01) and the reversal multiples were 2.59 and 1.69 respectively. After treating with 20 micromol/L and 10 micromol/L PDMP for 48 hours, the concentration of DNR in K562/A02 cells increased (p < 0.05) and the apoptotic rate also was elevated (p < 0.01). The expressions of GCS and mdr1 genes were down-regulated at mRNA and protein levels after treating K562/A02 cells with 20 micromol/L PDMP for 48 hours. It is concluded that PDMP can enhance the sensitivity of K562/A02 cells to DNR by increasing cell apoptosis rate and accumulation concentration of DNR in cells, which may be related to down-regulated expressions of GCS and mdr1 genes.
Daunorubicin ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Enzyme Inhibitors ; pharmacology ; Humans ; Inhibitory Concentration 50 ; K562 Cells ; Morpholines ; pharmacology

The aim of this study was to investigate the combined effects of bortezomib (Bor) and daunorubicin (DNR) or each drug alone on proliferation of human multiple myeloma cell line KM3. KM3 cells were cultured with different concentrations of Bor and DNR, Bor or DNR alone for different times. The cell proliferation was analyzed by MTT assay, and the concentration of 50% growth inhibition (IC(50)) was calculated. The results indicated that both of Bor and DNR inhibited KM3 cell proliferation in dose dependent manner. The IC(50) of both drugs were 0.27 micromol/L and 0.16 micromol/L respectively. The inhibiting rate of Bor plus DNR on KM3 cells was much higher than that of Bor (p < 0.05). It is concluded that the Bor has synergistic inhibitory effect with DNR on the growth of KM3 cell in vitro.
Boronic Acids ; pharmacology ; Bortezomib ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Daunorubicin ; pharmacology ; Drug Synergism ; Humans ; Inhibitory Concentration 50 ; Multiple Myeloma ; Pyrazines ; pharmacology

The aim of this study was to investigate the potential benefit of combination therapy with 5-bromotetrandrine (5-BrTet) and daunorubicin (DNR) on chronic leukemia. The apoptosis of K562/A02 cells treated by DNA, BrTet and BrTet combined with DNR for 48 hours was detected by flow cytometry; the expressions levels of survivin mRNA and protein K562/A02 cells treated by DNR, BrTet and BrTet combined with DNR and in untreated K562 cells for 48 hours were measured by RT-PCR and Western blot respectively. The results showed that the combination of BrTet with DNR increased apoptotic rate of K562/A02, down-regulated the expression levels of survivin mRNA and protein in K562/A02 cells as compared with blank control and cells treated by BrTet or DNR alone, the survivin expression in K562/A02 cells was higher than that in K562 cells. It is concluded that the combination of BrTet with DNR can effectively reverse the multidrug resistance of K562/A02 cells, promote the apoptosis of K562/A02 cells, the mechanism of which may be related with down-regulation of survivin expression. Survivin may be a target for the treatment of MDR in hematopoietic malignancies.
Apoptosis ; drug effects ; genetics ; Benzylisoquinolines ; pharmacology ; Daunorubicin ; pharmacology ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Gene Expression Regulation, Leukemic ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; K562 Cells

Azacitidine ; administration & dosage ; analogs & derivatives ; pharmacology ; Cell Proliferation ; drug effects ; Daunorubicin ; administration & dosage ; pharmacology ; HL-60 Cells ; Humans ; PTEN Phosphohydrolase ; metabolism

OBJECTIVETo investigate the effects of arsenic trioxide (As(2)O(3)) on the apoptosis and P-glyco-protein (P-gp) expression of multidrug-resistant human leukemia K562/ADM cells, and the combined effects of As(2)O(3) with conventional chemotherapeutic agents.

METHODSMultidrug-resistant human leukemia cell line K562/ADM that overexpresses mdr-1 gene was used as the target cells. The cell proliferating activity was assessed with a MTT assay. Cell morphology was examined by light microscopy, confocal microscopy and electron-microscopy. P-gp expression, cell-cycle status were determined by flow cytometry.

RESULTSK562/ADM cells were highly resistant to adriamycin, and cross-resistant to daunorubicin and etoposide. As(2)O(3) at concentrations of 0.5 to 20 micromol/L inhibited the proliferation of K562/ADM cells, and K562/ADM cells were more sensitive to As(2)O(3) than their parent K562 cells did. As(2)O(3) induced marked apoptosis of K562/ADM cells showed by typical apoptotic morphological changes and the appearance of high sub-G(1) cell population. As(2)O(3) significantly inhibited the P-gp expression in K562/ADM cells, and exerted a synergistic effect on the enhancement of the cell sensitivity to adriamycin, daunorubicin and etoposide.

CONCLUSIONAs(2)O(3) induces growth-inhibition and apoptosis of multidrug-resistant K562/ADM cells, and augments synergistically the sensitivity of the cells to conventional chemotherapeutic agents via down-regulation of P-gp expression.

ATP-Binding Cassette, Sub-Family B, Member 1 ; biosynthesis ; drug effects ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Daunorubicin ; pharmacology ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Drug Synergism ; Etoposide ; pharmacology ; Humans ; K562 Cells ; Oxides ; pharmacology