Effect of Arsenic Trioxide on Cell Cycle and Apoptosis of Multidrug Resistant K562/A02 Cell Line
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Objective: To explore the effect of arsenic trioxide on cell cycle and apoptosis of K562/A02 cells and its possible mechanism.Methods: Adriamycin(Adr) resistant K562/A02 were treated with arsenic trioxide(non-cytotoxic concentration at 4.0υmol/L,5.0υmol/L) or without arsenic trioxide(control),flow cytometry was used to evaluate apoptosis and cell cycle distribution,and change of the expression level of NF-κBp65 protein in nucleus was detected by western blot.Results: As compared with control arsenic trioxide significantly increased the rate of apoptosis,arrested cells in G0/G1phase and reduced the levels of NF-κBp65 protein in nucleus(allP0.05).Conclusion: The underlying mechanism for arsenic trioxide to promote apoptosis of K562/A02 and suppress cell proliferation lies in its impact on NF-κBp65 protein expression.Keywords:
Arsenic Trioxide
K562 cells
Trioxide
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Objective:To explore the effect of arsenic trioxide on cell apoptosis in HL-60 cells.Methods:DNA(deoxyribonucleic acid) line was detected by gelose electrophresis.Changes of cell nucleus stained by Hoecst 33258 in HL-60 cells treated by arsenic trioxide were studied by fluorescence microscope.The apoptosis were detected by flow cytometry.Results:DNA fragment appeared after HL-60 cells exposed to 10 μmol/L arsenic trioxide from 24 to 48 h;cell nucleus were concentrated and fragmented after HL-60 cells exposed to 10 μmol/L arsenic trioxide for 48 h.Treated with 10 μmol/L arsenic trioxide for 12hours,24 hours and 48 h,the apoptotic rate was (8.5±2.1)%,(12.8±3.4)% and(21.4±5.8)%,respectively,but the apoptotic rate in the negative control group only (1.5±0.5)%(P0.05).Conclusion:Arsenic trioxide can induce apoptosis in HL-60 cells.
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Trioxide
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Objective To investigate the inhibition effect of arsenic trioxide combined with interferon on the growth of epidermal carcinoma cells at different concentrations and different time in order to select the optimized action time and dose of the combination of the two drugs.Methods Through cell culture with human epidermal carcinoma cells A431 in vitro,the inhibition effects of arsenic trioxide combined with INF at different concentrations on proliferation of A431 cells were detected by MTT.The apoptosis rate and the changes of cell cycle in human A431 cells were detected by FCM.Results The results of MTT showed that arsenic trioxide and INF respectively inhibited the proliferation of A431 cells in a dose-dependent and time-dependent manner.The combination of arsenic trioxide with INF inhibited the growth of A431 cells,and the inhibitory rate of the combination application was significantly higher than that of arsenic trioxide only,there was a significant difference between arsenic trioxide and INF(P0.01).There was a synergistic effect of arsenic trioxide and INF combination application.The synergistic effects of arsenic trioxide and INF on inducing apoptosis of A431 cells was more obvious,as compared with that of using simple arsenic trioxide or INF.Conclusion The arsenic trioxide and INF has inhibition effect on the growth of A431 cells in vitro respectively in a dose-dependent and time-dependent manner.The arsenic trioxide and INF can result in the arrearage of S phase of cell growth,and can increase the ration of S phase,and there is a synergistic effect of arsenic trioxide and INF combination application.The induction effect of arsenic trioxide on apoptosis of A431cells is quite obvious,nut INF can enhance the induction effect of arsenic trioxide on apoptosis of A431cells.
Arsenic Trioxide
A431 cells
Trioxide
MTT assay
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Objective:To study the effect of arsenic trioxide on induction of apoptosis of melanoma A 375 cell line.Methods:Early apoptosis rate and late apoptosis rate in A 375 cells were analysed by flow cytometry.Results:After pretreated with arsenic trioxide of 5 μmol/L,10 μmol/L,and 20 μmol/L,the early apoptosis rate in A 375 cells were 9.75%,50.9%,and 43.7%,respectively.The late apoptosis rate in A 375 cells were 2.10%,4.61%,and 11.2%,respectively.The totle apoptosis rate were 11.85%,55.51%,and 54.9%,respectively.Conclusions:The early and the late apoptosis rate in A 375 cells were induced by arsenic trioxide.
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Objective To explore the changes of Par-4 and WT1 genes expression during human leukemic K562 cells apoptosis induced by arsenic trioxide (As2O3). Methods After the K562 cells were treated with arsenic trioxide (2-10 μmol/L) for 24-72 hours, cell survival rate was evaluated by MTT assay and apoptosis was analyzed using flow cytometry. The expressions of mRNA and protein for par-4 and WT1 were tested by Real-time quantitative reverse transcriptase polymerase chain reaction (RQ-RT-PCR) method and Western blotting in K562 cells with various concentrations of arsenic trioxide at different time points. Results After the K562 cells were treated with As2O3 of different concentrations, arsenic trioxide could efficiently inhibit the growth of K562 cells and induce apoptosis with the increase of As2O3 concentration. The same time, the mRNA and protein expressions of Par-4 were up-regulated, while the mRNA and protein expression of WT1 were down-regulated. Conclusion Arsenic trioxide could inhibit the growth of K562 cells and induce apoptosis. Par-4 and WT1 genes could participate in the apoptosis of K562 cells induced by arsenic trioxide.
Key words:
Arsenicals; K562 cells; Apoptosis; Tenes, Par-4; Genes, Wilms tumor
Arsenic Trioxide
K562 cells
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Objective To study the effect of arsenic trioxide on proliferation,apoptosis,cell cycles and phosphorylation oncogene(PPO) expression in the human melanoma cell line A375.Methods The human melanoma cell line A375 was cultured.MTT assay was used to detect the effect of arsenic trioxide on proliferation, apoptosis and cell cycles of A375 cells at different concentrations.The effects of arsenic trioxide on cell cycle and apoptotic rates were assessed by flow cytometry.The morphological changes of cells were determined by the reverse microscopy after the treatment with arsenic trioxide.The influence of arsenic trioxide on the expressions of PPO in the A375 cells were detected with immunohistochemical method.Results It was confirmed by MTT assay that within the concentration interval of 1~10 μmol/L,as the concentration and exposure time increased,the inhibition ratio of arsenic trioxide on the A375 cells gradually increased.The inhibitory effects of arsenic trioxide on the proliferation of the A375 cells showed a time-and concentration-dependent manner.The growth of the A375 cells were arrested at S stage after treatment with arsenic trioxide with flow cytometry.It could be seen by the reverse microscopy that the typical morphological change of apoptosis occurred in the A375 cells.PPO protein was expressed in the intracytoplasm of the A375 cells.The expression of PPO protein diminished in arsenic trioxide-treated cells compared with the controls.There was difference between the control and the experiment groups(P0.05).Conclusions Arsenic trioxide significantly inhibits the proliferation of the A375 cells and there is a positive correlation between the concentration and effectiveness of the drug.The mechanism of action might be related to the apoptosis resulting from inhibition of PPO expression in the A375 cells.
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MTT assay
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To explore the mechanism of arsenic (arsenic trioxide, AS2O3) in treating leukemia.The in vitro effect of arsenic trioxide on HL-60 model double-labelled with PI/AnnexinV-FITC was studied ith flow cytometry. Function on telomerase activity was also observed.After the effect of arsenic on HL-60 cell line, PI negative/AnnexinV-FITC positive cells increased, it mainly acts at the G2-M phase that suggests the apoptosis takes place, mild inhibition on telomerase was also found 48 hours later.Apoptosis induced by arsenic trioxide was the main mechanism in killing or inhibiting leukemic cells.
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Objective To investigate the effects of arsenic trioxide (As 2O 3) on the cell cycle and apoptosis of human nasopharyngeal carcinoma cell line. Method Cell morphology, flow cytometry and TUNEL were used to examine the cell apoptosis of human nasopharyngeal carcinoma cell line CNE1 indued by arsenic trioxide.The cell cycle was determined by using flow cytometry. Result Arsenic trioxide induced CNE 1cell apoptosis with typical morphological features, and the apoptotic peak detected by flow cytometric analysis showed a dose-dependent effect. FCM data showed that the G 2/M phase ratios were increased significantly in a dose-dependent manner.Conclusion As 2O 3 could significantly induce CNE1 cell lineapoptosisand alter the cell cycle progression.
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Objective The current study was designed to investigate the effect of arsenic trioxide(As_(2)O_(3)) on induction of apoptosis in ovarian cancer cell.Methods Cell proliferation was assessed by MTT assays,morphological changes of apoptosis were observed with invert microscope and transmission electron microscope.DNA ladder and cell cycle were examined by DNA agarose gel eletronphores and PI fluorescence flow cytometry(FCM)respectively.Results Under the influence of arsenic trioxide,HO8910 cell exhibited changes of apoptosis both in morphology and in the characteristics of electrophoresis.Meanwhile the cell cycle also showed special change with the apperance of G1 peak and increase of S+G2~M cells.Conclusion Arsenic trioxide inhibits cell proliferation,induces apoptosis in ovarian cancer cell.
Arsenic Trioxide
Agarose gel electrophoresis
MTT assay
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Objective:To study the changes of cell cycle phase and cell cycle phase regulation protein during arsenic trioxide induced apoptosis in hepatoma cells,and to explore mechanisms of arsenic trioxide inhibited HCC.Methods:Cultured in vitre,HCC SMMC-7721 were treated 72h with 2μmol/L arsenic trioxide.The cell cycle and apoptosis index were detected by flow cytometry(FCM);the morphological changes were observed by transmission electron microscope.The immunohistochemistry was used to detect the protein expression of p21、cyclin D_1,cyclinA.Results:SMMC-7721 cell cycle was arrested in G_1 and S phase by arsenic trioxide.Compared with control in the course of apoptosis induced by arsenic trioxide p21 protein expressions rose significantly(control:1.128;arsenic trioxide:3.794),while cyclinD_1,cyclinA protein expressions decreased significantly(control 1:3.647,2.833;arsenic trioxide:1.133,1.179).Conclusion:Arsenic trioxide could disturb cell cycle progression of SMMC-7721 and induce apoptosis.The mechanism may relate to that arsenic trioxide up-regulate expression of p21 and down-regulate expression of cyclinD_1,cyclinA.
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To investigate the anti-proliferation effect of arsenic trioxide on lung cancer A549cells and its mechanism.A549cells were cultured in different concentrations of arsenic trioxide medium in vitro.The inhibitory ratio of the cells were measured by MTT assay.Apoptosis was observed by flow cytome-try(FCM)and Hoechst33258fluorescence staining.The activity of telomerase was detected by TRAP-PCR-ELISA before and after apoptosis occurred.Arsenic trioxide could decrease the telomerase activity,cause apoptosis and inhibit the growth of A549cells significantly.The suppression was both in time-depen-dence and dose-dependence.Morphological changes including condensation of celluar chromatin and frag-mentation of nuclear were found in Hoechst33258fluorescence staining.[Conclusion]Arsenic trioxide can in-hibit the growth of A549cells and induce apoptosis.Telomerase activity reduced in A549cells may be one of the important mechamisms.
Arsenic Trioxide
Trioxide
MTT assay
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