Effect of vitamin K_(3) on apoptosis induced by androgen - independent prostate cancer cell PC-3M
0
Citation
0
Reference
20
Related Paper
Abstract:
AIM:To study the effect of vitamin K3(VK3) on the induction of apoptosis in androgen-independent prostate cancer cell PC-3M in vitro.METHODS:Cell viability was estimated by MTT assay.AO/EB staining was performed to detect apoptotic cells.Apoptosis and the changes of cell cycle were detected by flow cytometry.NAC was used to observe the effect of growth inhibition by VK3.RT-PCR was used to confirm the changes in gene expression.Levels of intracellular peroxides were estimated by using an oxidation-sensitive fluorescent probe DCFH-DA.RESULTS:PC-3M cells growth was significantly inhibited by VK3(≥60 μmol/L,P0.05).The inhibitory effect was time and dosage dependent.The result of AO/EB staining showed that apoptosis of PC-3M cells were induced by VK3.A typical subdiploid peak before G0/G1 phase was observed after treated for 12 h with VK3(60 μmol/L) by flow cytometry.The effect of growth inhibition treated with VK3 was antagonized by antioxygen NAC(5,10,20,40,80 μmol/L).An increase in the level of DCF fluorescence after PC-3M cells were treated for 1-2 h with VK3 was observed.Antioxidase GSH-Px and CAT were run-down after treated with VK3.CONCLUSION:The results indicate that apoptosis in PC-3M cells is induced through oxidative stress by VK3.Keywords:
Viability assay
Growth inhibition
Topics:
Cite
Objective To study the effects of quercetin(QUE) on proliferation of rat glioma C6 cell line in vitro.Methods The cells were divided into 5 treatment groups(10,25,50,75 and 100 μmol·L~(-1) QUE),blank control and menstruum control group.The rat C6 cells were cultivated to 1×10~6·mL~(-1) in the RPMI 1640 medium,then added into 96 holes board with various doses of QUE by 3 holes per group,and MTT assay was used to observe the proliferation of the cells treated for 24,48 and 72 h.The change of cell cycle was also observed by flow cytometry(FCM) after the cells were treated with 50 and 100 μmol·L~(-1) QUE for 48 h.The changes of the protein P53 and Bcl-2 of C6 cells treated with 50 μmol·L~(-1) QUE for 48 h were detected by immunocytochemical methods.(Results With) the augmentation of QUE and the extension of the treated time,the C6 cell growth was inhibited,the A values decreased and the cell number in G_0/G_l phase was increased,the cell numbers in S and G_2/M phases were cut down,and the decreased expression of Bcl-2 protein and the increased expression of P53 protein were also observed after treatment with QUE.Conclusion Inhibitory effect of QUE on C6 cell line is proved to be dependent on the treated time of the drug and the dose of QUE,and the induced apoptosis of C6 cells is implemented by the means of up-regulation of P53 protein expression and down-regulation of Bcl-2 protein expression.
Cell counting
MTT assay
Cite
Citations (3)
Objective: To invesfgate the effects of various concentrations As2O3 on malignant melanoma. Methods: The viability of Cloundman melanoma S91 cells treated with As2O3 was measured by MTT [3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide] assay. The apoptosis was determined by transferase-mediated dUTP nick end labeling (TUNEL) detection, and the morphology of apoptotic cells was observed through transmission electron microscopy (TEM). The cells growth phase were analyzed by flow cytometry (FCM). Results: A time-and dose-dependent decrease in cell viability was induced in S91 cells after treatment with As2Os at the concentration of 1-5 μmol/L respectively. The TUNEL indices were 0.033 ± 0.018, 0.062 ± 0.012, 0.102 ± 0.016, 0.132 ± 0.031,and 0.162 ± 0.027 respectively, which were much higher compared with the control group (0.017 ± 0.004, P ＜ 0.01). The flow cytometry showed that hypodiploid peak after treatment with 3 μmol/L and 5 μmol/L of As2O3 for 48 h were 9.99% and 17.59% respectively,which increased significantly compared with the control cells (3.05% P ＜ 0.01, ). The apoptotic morphology observed by transmission electron microscope showed the chromatin became condensed and attached to the inner surface of nuclear membrane. Conclusion: Arsenic trioxide can induce melanoma Cloundman S91 cell apoptosis at the concentration of 1-5 μmol/L, which will provide enhanced benefit in melanoma therapy.
Arsenic Trioxide
Viability assay
Cite
Citations (0)
AIM: To induce preconditioning and oxidative stress by H_2O_2 in HepG2 cells. METHODS: The different doses of H_2O_2 were used to induce apoptosis in HepG2 cells, which was estimated by AO/EB staining, MTT assay and flow cytometry. RESULTS: The different group of HepG2 cells stained with AO/EB showed different staining state. The high dose of H_2O_2 resulted in the increase in apoptosis rate of HepG2 cells and made MTT activity decreased. However, after pretreated with low dose of H_2O_2, the apoptosis rate was decreased and MTT activity was increased. CONCLUSION: The high dose H_2O_2 induced apoptosis in HepG2 cells and the low dose H_2O_2 protected HepG2 cells against the oxidative stress.
MTT assay
Cite
Citations (0)
Objective:To study the effects of vitamin C on the growth of human cervical carcinoma cell line,HeLa cells,and on the generation of reactive oxygen species (ROS) in the cells. Methods: HeLa cells were treated with various concentrations of vitamin C.The effect of vitamin C on HeLa cells survival and apoptosis was determined by MTT assay,light microscope and flow cytometry,and cellular ROS was measured by fluorometer. Results: After being treated with 0.25 mmol/L vitamin C for 24 h,the survival of HeLa cells did not change.However,when cells were treated with vitamin C in 0.5-4 mmol/L,the survival of cells were significantly decreased,a marked apoptosis characteristic was observed in time-and dose-dependent manner.When we measured the cellular hydrogen peroxide (H2O2) level we found that the H2O2 level in HeLa cells started to rise after being incubated with 0.5-4 mmol/L vitamin C for 15 min,and sustained increased,reached the peak point of H2O2 level at 30 min,and maintained a little time,then it began to decrease.While cells incubated with vitamin C in 0.5-4 mmol/L for 1 h to 24 h,the intracellular H2O2 level were 50% of that of the control group. Conclusion: The higher concentration of Vitamin C could induce the apoptosis on HeLa cells,derived from the solid tumors,the molecular mechanisms of which may be related to intracellular ROS level.
HeLa
MTT assay
Cite
Citations (0)
Experimental Study on Activation of Caspase-3 and Apoptosis of K562 Cell Induced by Iron-Deprivation
Objective To observe the relationship between apoptosis of K562 cell induced by iron-deprivation and activation of Caspase-3.Methods K562 cells were treated with desferrioxamine(DFO) in different dosages were collected at different time points.K562 cells were labelled with Annexin V/PI,and then the rate of apoptosis was measured by flow cytometry;The activation of Caspase-3 were detected by colorimetric method with pAN labelled substrate;The active protein of Caspase-3 were analyzed by Western blot.Results When K562 cells were treated with different concentrations of DFO,the apoptosis rate and the activity of Caspase-3 increases gradually.When K562 cells were incubated with DFO(50 μmol/L and 100 μmol/L) 24 h later,the enzymatic activity of Caspase-3 increases dramatically more than that of control group,and the difference was significantly(P0.001);The active protein of Caspase-3 could been found when K562 cells were treated with DFO(100 μmol/L) 14 h later,the activity and the quantity of Caspase-3 depends on time-dosage;Compared with control group at 10 h and 12 h time point,the activation of Caspase-3 was not different significantly from each other(P0.05).All those effect above can be counteracted by equal mole concentration of FeCl_3.Conclusion Iron-deprivation maybe induce the apoptosis of K562 cell by chelating intracellular iron and activing Caspase-3.
K562 cells
Caspase-9
Caspase 12
Cite
Citations (0)
AIM:To explore the proliferation inhibition and apoptosis inducing effects of matrine (Mat) on U937 cells and its mechanism. METHODS:The proliferation inhibition rate of U937 cells was assayed by MTT analysis; cell cycle was evaluated by flow cytometry (FCM); cell apoptosis rate was calculated by Annexin/PI double staining method. The Bcl-2 mRNA expression of U937 cells treated with Mat was assayed by RT-PCR. RESULTS:After U937 cells were treated with Mat (0.2-0.5 g/L) for 24,48 and 72 h,the cell proliferation was markedly inhibited in a dose-dependent and time-dependent manner (P0.05 or P0.01),and the 5% inhibiting concentration (IC50) was 0.4 g/L. Cell cycle analysis showed that it was arrested in the S phase after 72 h. When the U937 cells were treated by Mat (0.2-0.5 g/L) for 72 h,the corresponding apoptosis rate (3.25%,5.32%,8.17%,11.20%) indicated by AnnexinV-FITC double staining,had statistical significance compared with the control group (1.84%) and the 0.1 g/L Mat-treated group (1.95%,P0.05 or P0.01). The Bcl-2 mRNA expression in U937 cells was decreased 72 h after treated by Mat (0.2-0.5 g/L) with a concentration-effect relationship. CONCLUSION:Mat can suppress the U937 cell proliferation in a concentration-dependent and time-dependent manner. The effect has a relation with the U937 cell cycle arrested in the S phase. Mat can down-regulate the expression of Bcl-2 and induce the apoptosis.
U937 cell
Cite
Citations (0)
To investigate the effects of trimethyltin chloride (TMT) on proliferation, apoptosis, oxidative damage, and NF-κB expression in PC12 cells in vitro.PC12 cells were treated with 0, 0.3125, 0.6250, 1.2500, 2.5000, 5.0000, 10.0000, and 20.0000 µmol/L TMT for 24 and 48 h, and MTT assay was used to evaluate cell viability. PC12 cells were treated with 1.25, 2.50, 5.00, and 10.00 µmol/L TMT for 12 and 24 h, and flow cytometry was used to measure the apoptotic rates of cells. PC12 cells were treated with 1.25, 2.50, 5.00, and 10.00 µmol/L TMT for 6 h, and the reactive oxygen species (ROS) and glutathione (GSH) levels were measured. PC12 cells were treated with 1.25, 2.50, 5.00, and 10.00 µmol/L TMT for 12 h, and Western blot was used to measure NF-κB levels.Compared with solvent controls, the PC12 cells treated with 2.5000, 5.0000, 10.0000, and 20.0000 µmol/L TMT for 24 h showed significantly decreased cell viability (P < 0.05); the PC12 cells treated with 1.2500, 2.5000, 5.0000, 10.0000, and 20.0000 µmol/L TMT for 48 h showed significantly decreased cell viability (P < 0.05). The PC12 cells treated with 1.2500, 2.5000, 5.0000, and 10.0000 µmol/L TMT for 12 h had apoptotic rates of 15.30% ± 0.75%, 18.90% ± 0.61%, 22.00% ± 0.60%, and 36.50% ± 0.66%, respectively, and the PC12 cells treated with 1.25, 2.50, 5.00, and 10.00 µmol/L TMT for 24 h had apoptotic rates of 28.6% ± 0.40%, 43.54% ± 2.00%, 65.73% ± 0.71%, and 74.67% ± 0.40%, respectively, all significantly higher than those of the control group (12 h: 12.80% ± 1.00%, 24h: 16.83% ± 0.25%) (P < 0.05). The ROS fluorescence intensities of the PC12 cells treated with 1.25, 2.50, 5.00, and 10.00 µmol/L TMT were 1.42, 1.71, 1.78, and 1.89 times that of the control group (P < 0.05); the PC12 cells treated with 2.50, 5.00, and 10.00 µmol/L TMT had GSH levels of 0.17 ± 0.0, 0.20 ± 0.04, and 0.07 ± 0.03 µmol/µg protein, significantly lower than that of the control group (0.30 ± 0.01 µmol/L protein) (P < 0.05). The PC12 cells treated with 2.50, 5.00, and 10.00 µmol/L TMT had significantly higher expression of NF-κB p65 than the control group (P < 0.05).Under our laboratory conditions, TMT can significantly inhibit proliferation and induce apoptosis in PC12 cells, which may be related to oxidative stress and NF-κB signaling pathway activation.
Viability assay
MTT assay
Cite
Citations (2)
OBJECTIVE: To investigate the proliferation and apoptosis of human gastric cancer SGC7901 cell induced by ApoG2 in vitro.METHODS: By using 3.125,6.250,12.500,25.000 and 50.000 μmol/L concentrations of ApoG2 to treat SGC7901 cells for 24,48 and 72 h,the inhibitory rate of cell proliferation was assessed by MTT method.After 24 h treated with 10,20 and 30 μmol/L ApoG2,MGG staining was used to investigate the cells morphological changes.Flow cytometry(FCM) was used to determine the cellular apoptosis with 10,20,30 and 40 μmol/L ApoG2.The expressions of Bcl-2,Bax and NF-κB mRNA were measured by RT-PCR with 10,20 and 30 μmol/L ApoG2.RESULTS: The inhibitory rate of SGC7901 cells treated with ApoG2 were higher than those in the negative control group(P0.05) with concentration-dependent,and the half inhibitory(IC50) were 32.58,25.11 and 14.16 μmol/L at 24,48 and 72 h;With the increasing of drug concentration,cells became slow,cytoplasm became loose,nuclei were darker,nucleoplasm ratio increased and cells presented typical cells death form.Apoptosis could be seen under flow cytomtry,after 24 h of ApoG2,early apoptosis rates were(1.92±0.55)%,(3.36±0.55)%,(5.07±0.70)%,(7.44±1.47)% and(7.88±1.22)%,and late apoptosis rates were(2.80±0.86)%,(13.09±0.93)%,(16.48±1.03)%,(18.32±1.44)% and(25.49±1.59)%,with the increase of ApoG2 with 0,10,20,30 and 40 μmol/L,the apoptosis rate of SGC7901 cells was increased(P0.01);The RT-PCR result showed that the expression levels of Bcl-2 and NF-κB mRNA were decreased(P0.05) with the increase of ApoG2,while the expression of Bax mRNA was up-regulated(P0.05).CONCLUSION: ApoG2 can inhibit the proliferation of SGC7901 cells and induce the apoptosis in vitro.
IC50
Nucleoplasm
Cite
Citations (0)
Objective To explore the inducing effect of vitamin K3 on apoptosis of human bladder cancer cell line T24.MethodsT24 cells were cultured in the presence of different concentrations of VK3(0,2,5,10,20 μmol/L).The inhibitory rates of VK3 were detected by MTT colorimetry,and apoptosis was observed using flow cytometry.ResultsAfter T24 cells were cultured with VK3 for 48 h at concentrations of 0,2,5,10,20 μmol/L,the inhibitory rates of VK3 were(5.80±0.038)%,(13.30±0.036)%,(26.55±0.032)%,(42.58±0.028)% and(65.35±0.034)%,respectively,and the apoptotic rates were(2.86±0.4)%,(5.16±0.7)%,(10.85±1.1)%,(23.01±0.8)% and(75.80±1.2)%,respectively.Cell growth significantly decreased in the presence of 10 μmol/L,20 μmol/LVK3(P0.05).ConclusionVK3 at the concentrations of 10-20 μmol/L could obviously inhibit the proliferation of T24 cells and induce apoptosis after 48 h culture in vitro.
MTT assay
Colorimetry
Mole
Cite
Citations (0)
AIM:To investigate the effect of Cpd5 on the proliferation inhibition and apoptosis induction in human ovarian cancer SKOV3 cells.METHODS:The growth inhibition of Cpd5 in SKOV3 cells was detected by MTT assay.AnnexinV/PI staining was employed for quantifying apoptotic cells by fluorescence microscopy and flow cytometry.The apoptotic morphology was observed by Hoechst-33258 staining.RESULTS:After Cpd5 treatment at the concentrations of 5,10,20,30,40,50 and 60 μmol/L for 48 h,the ratios of the cells growth inhibition were 2.77%,5.19%,10.61%,41.15%,71.37%,82.90% and 89.81%,respectively.The proliferation was inhibited after Cpd5 treatment at different times of 12,24,48 and 72 h,the cells were inhibited by Cpd5,in a dose-time-dependent manner.The apoptotic cells accounted for 9.25%,20.07% and 56.16%,after Cpd5 treatment by 30 μmol/L for 12,24 and 48 h,respectively.The ratio of apoptotic cells in 50 μmol/L group was significantly higher than that in 30 μmol/L group.The morphological changes were emerged after Cpd5 treatment at the concent rations of 40,50 and 60 μmol/L for 12 h.Compared with the control group,the ratios of the apoptotic cells were increased significantly in the groups induced by Cpd5 at different concentrations.The apoptotic cells were increased in the groups of Cpd5 treatment at 20 μmol/L for 24 h and 48 h by Hoechst-33258 staining.Furthermore,the ratios of apoptotic cells were increased significantly in the groups of Cpd5 treatment at 30,40,50 and 60 μmol/L Cpd5 at different time-points.CONCLUSION:Cpd5 can induce proliferation inhibition and apoptosis in SKOV3 cells,in a dose-time-dependent manner.Our data reveal that Cpd5 is a novel anti-ovarian cancer compound.
MTT assay
Growth inhibition
Cite
Citations (0)