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OBJECTIVE@#To observe the expression of c-Myc protein in cervical cancer HeLa cells and explore the effect of juglone on the proliferation and apoptosis of HeLa cells by affecting c-Myc ubiquitination.@*METHODS@#HeLa cells treated with different concentrations (0, 10, 20, or 50 μmol/L) of juglone or with 20 μmol/L juglone for different time lengths were examined for expression of c-Myc protein with Western blotting. The half-life of c-Myc protein was determined using cycloheximide (CHX) and c-Myc protein degradation was detected using coimmunoprecipitation. We also assessed the effects of 20 μmol/L juglone combined with 0, 1.0 or 2.0 μmol/L MG132 (a proteasome inhibitor) on c-Myc expression. The effects of 20 μmol/L juglone on the proliferation and apoptosis of HeLa cells with RNA interference-mediated knockdown of c-Myc were evaluated with MTT assay and flow cytometry.@*RESULTS@#Treatment with juglone significantly lowered c-Myc protein expression in HeLa cells in a concentration-and time-dependent manner (P < 0.05). Juglone obviously shortened the half-life of c-Myc protein, and the addition of MG132 significantly up-regulated the expression level of c-Myc protein (P < 0.05). Juglone treatment also promoted ubiquitination of c-Myc protein in HeLa cells. Compared with the cells transfected with a negative control construct, the cells transfected with si-c-Myc showed significantly decreased proliferation inhibition and a lowered cell rate with early apoptosis after juglone treatment (P < 0.05).@*CONCLUSION@#Juglone inhibits proliferation and promotes apoptosis of HeLa cells by affecting the ubiquitination of c-Myc protein.
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Female , Humans , Apoptosis , Cell Proliferation , HeLa Cells , Naphthoquinones , Ubiquitination , Uterine Cervical Neoplasms/geneticsABSTRACT
OBJECTIVE：To investigate th e effects of juglone on brain tissu e of rats and its relationship with the biomarkers related to brain tissue injury. METHODS ：Totally 40 rats were divided into blank group ，juglone high-dose ，medium-dose and low-dose groups （34.832，17.416，8.708 mg/kg），with 10 rats in each group. They were given medicine intragastrically once a day ， for consecutive 4 weeks.After last administration ，the general behavior ，brain index and brain tissue morphology were investigated. UPLC-MRM-MS method was used to determine the serum contents of L-dopa（L-Dopa），L-tyrosine（L-Tyr）and L-tryptophan（L-Trp） in rats. The chromatographic condition included Waters Acquity UPLC BEH C 18 column，mobile phase consisted of ammonium acetate aqueous solution-acetonitrile ，gradient elution ，at the flow rate of 0.3 mL/min，sample size of 5 μL，column temperature of 30 ℃；MS condition include electrospray ion source ，in positive ion mode ，capillary voltage of 3 500 V，desolvent gas flow of 650 L/h，desolvent temperature of 350 ℃，ion source temperature of 110 ℃. RESULTS ：Compared with blank group ，the rats in each dose group showed the behavior of tiredness and weakness of limbs. The brain tissue morphology showed pathological changes ， which contained blood vessel congestion in the cerebral and cerebellar cortex ，partial cell nucleus pyknosis in the pyramidal cell layer，deep staining of nuclei ，irregular shape and unclear boundary and other pathological changes ；the brain index of juglone high-dose group increased significantly （P＜0.05）. The established UPLC-MRM-MS method showed good specificity and the range of L-Dopa，L-Tyr and L-Trp were 31.25-32 000，31.25-32 000，15.625-16 000 ng/mL（r＝0.999 1-0.999 9），respectively. The limits of detection were 6.250，5.625，3.125 ng/mL，respectively. The limits of quantitation were 15.625，18.75，10.00 ng/mL，respectively. RSDs of precision ，accuracy and stability （24 h）tests were all Matrix effects were 95.1％-100.1％ （RSD are not more than 3.25％，n＝3）. Compared with blank group，the contents of L-Dopa were increased significantly injuglone medium-dose and high-dose groups （P＜0.01）. The contents of L-Tyr were increased significantly in juglone lowdose，medium-dose and high-dose gro ups，while the contents of L-Trp were decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS ：Juglone has an effect on the general behavior，brain index and brain tissue morphology of rats. It may affect the brain function of rats by increasing the contents of L-Dopa and L-Tyr in serum and decreasing the contents of L-Trp.
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BACKGROUND: Juglone is a naphthoquinone currently obtained by chemical synthesis with biological activities including antitumor activity. Additionally, juglone is present in the green husk of walnut, which suggests evaluating the effect of GH extracts on carcinogenic cell lines. RESULTS: Walnut green husk ethanolic extract was obtained as 169.1 mg juglone/100 g Green Husk and antioxidant activity (ORAC) of 44,920 µmol Trolox Equivalent/100 g DW Green Husk. At 1 µM juglone in HL-60 cell culture, green husk extract showed an antiproliferative effect, but pure juglone did not; under these conditions, normal fibroblast cells were not affected. A dose-dependent effect on mitochondrial membrane potential loss was observed. Apoptosis of HL-60 was detected at 10 µM juglone. Despite high ORAC values, neither purified juglone nor the extract showed protective effects on HL-60 cells under oxidative conditions. CONCLUSIONS: Green husk extract generates an antiproliferative effect in HL-60 cells, which is related to an induction of the early stages of apoptosis and a loss of mitochondrial membrane potential. The normal cells were not affected when juglone is present at concentrations of 1 µM, while at higher concentrations, there is loss of viability of both cancerous and healthy cells.
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Apoptosis , HL-60 Cells/metabolism , Juglans/chemistry , Polyphenols/metabolism , Antioxidants/metabolism , Cell Survival , Chromatography, High Pressure Liquid , Cell Culture Techniques , Membrane Potential, MitochondrialABSTRACT
Naphthoquinones have protective effects through different mechanism against to human malignancies including pancreatic cancer. One of these mechanisms is to avoid reactive oxygen species (ROS) production. Changes in enzymatic (superoxide
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Background: Colon cancer is a major cause of morbidity and mortality in the world. Juglone is a natural compound which has been isolated from Juglans mandshurica Maxim, and it has various pharmacological effects such as antiviral, antibacterial, and anticancer. In our study, we aimed to investigate the effect of juglone on CCL-228-SW 480 colon carcinoma cell line in monolayer and spheroid culture medium. Materials and Methods: The CCL-228-SW 480 cell lines were cultured in both monolayer and spheroid cultures. Cells were treated with juglone at 24, 48, and 72 h of incubation. ID50 inhibition was determined on the dose for juglone and after it was found 20 μM was applied to the cells to examine the effect of juglone on CCL-228-SW 480 colon carcinoma cell line. After Juglone was applied the BrdU marking index, Transferase dUTP Nick ends Labeling (TUNEL) assay, active caspase-3 assay, apoptosis-inducing factor (AIF) assay were determined by immunohistochemistry in both the monolayer and spheroid cultures. Results: The control group had a healthy pattern of S-phase fraction, and many of the CCL-228-SW 480 cells nuclei were observed to be positive for BrdU. Terminal Deoxynucleotidyl TUNEL-positive cells, active Caspase-3, and AIF were detected after treatment with juglone in both the monolayer and spheroid cultures. Conclusions: The dead cell count was higher in the CCL-228-SW 480 cell lines with juglone applied than in the controls. Juglone significantly inhibits the proliferation and induces the apoptosis of CCL-228-SW 480 cells in vitro
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Objective: To prepare Juglone-loaded poly lactic-co-glycolic acid nanoparticles (Jug-PLGA-NPs), and investigate their physicochemical properties, release characteristics in vitro and anti-tumor activities on A375 melanoma cells in vitro. Method: Jug-PLGA-NPs were prepared by emulsification-solvent evaporation method. Then the particle size, encapsulation efficiency, drug loading rate and in vitro release characteristics were investigated. Fluorescence microscopy was used to observe the uptake of PLGA-NPs in vitro. The distribution of PLGA-NPs in BALB/c nude mice after tail vein injection was observed by the small living animal imaging system. Their inhibition effect on proliferation of A375 cells was detected by thiazolyl blue tetrazolium bromide (MTT) assay. Apoptosis rate and cell cycle detection were performed by flow cytometry. Western blot was used to determine the protein kinase B (Akt), phosphorylated Akt (p-Akt) and cyclinD1. Result: The average particle size of the prepared Jug-PLGA-NPs was (149.6±21.5) nm, entrapment rate of (68.39±2.51)%, and drug-loading rate of (5.07±0.98)%, showing good sustained-release characteristics. PLGA-NPs showed good penetration and targeting properties in cellular uptake in vitro and in vivo imaging. Different concentrations of Jug-PLGA-NPs could significantly inhibit the proliferation and promote apoptosis of A375 cells in a time and concentration dependent manner (P1 expression (P0/G1 phase (PConclusion: The Jug-PLGA-NPs are easy to prepare and have good sustained-release characteristics, tumor targeting and anti-tumor ability, providing a new pharmaceutical dosage form for the future clinical application of Jug.
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Aims: To evaluate the effect of salicylic acid (SA) and acibenzolar-S-methyl (ASM) on the minimal concentration of juglone (Cmin) inducing foliar necrosis- and to determine the total phenol content and antioxidant activity of extracts of banana leaves after SA and ASM applications and toxin injection. Methodology: Banana cultivars Orishele and Corne 1 were subjected to root and foliar applications of elicitors, then leaves were injected with juglone a toxin of M. fijiensis to the determination of Cmin. The determination of total phenols extracted from leaves was carried out using Folin Reagent. The antioxidant activity of phenolic crude extracts (PCE) was determined through the DPPH radical scavenging ability. Results: In banana without elicitor applications, the minimum concentration inducing necrosis varied between 12.5 and 25 ppm of juglone but reached 250 ppm of juglone into banana treated with elicitors, particularly with ASM at 50 ppm. The phenol contents were highest 14 days after elicitors application. After this incubation time, with ASM at 50 ppm and AS at 25 ppm, the levels were 16.25 mg GAE/g DW in Orishele and 17.20 mg GAE/g DW in Corne 1, respectively. But the levels were lower 28 days after banana elicitation with 50 ppm of SA into Orishele and Corne 1 respectively 5.60 and 6.79 mg GAE/g DW. Treatments with 50 ppm of ASM showed the highest antioxidant activity between 7 and 28 days after elicitors application on banana leaves. With this treatment, the lowest concentration of phenolic crude extract scavenging 50% of DPPH was 13.09 μg/mL at 21 days after foliar elicitation. Conclusion: The applications of elicitors SA and ASM affect phenol content and antioxidant activity for the detoxification of foliar tissues of banana cultivars Orishele and Corne 1 infiltrated with toxin.
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Objective To optimize the best macroporous resin through static adsorption and desorption and establish enrichment process of juglone-constituents from Qinglongyi with the transfer rate of juglone constituents as the index. Methods The static adsorption and desorption of four different resins with different polarity to juglone-constituents from Qinglongyi were compared. AB8 resin was selected as the best resin. Results The craft parameters of the best resin were optimized, and the results were as follows: initial concentration was 0.0760 g/ml, the pH was 2.0, the flow rate of adsorption was 3BV/h, the elution concentration of ethanol was 70%, the elution volume was 3BV,the desorption flow rate was 2BV/h, and the ratio of diameter to height was 1:10. After the process of optimization, the content of juglone- constituents from Qinglongyi increased from 1.026% to 11.08% and the transfer rate was 72.08%. Conclusion The AB8 macroporous resin could be used to enrich and purify juglone-constituents from the Qinglongyi.
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Objective: To observe the effect of Juglone on invasion and metastasis of Hela cells and explore the possible mechanism. Methods: HeLa cells were cultured and treated with 10,20,50,100μmol/L Juglone for 24 hours. The morphology changes of HeLa cells were observed with an inverted microscope. The viability of HeLa cells was detected by MTT assay. The cell scratch test was used to detect cell migration ability after treatment of Juglone. The ability of cell invasion was measured by Transwell chamber. The expression of matrix metalloprateinases (MMP)-2 and MMP-9 were detected by Western blotting. Results: Compared with control group, the viability of HeLa cells decreased after treatment with different concentrations of Juglone for 24 hours, and the cell morphology was changed in a dose-dependent manner. Scratch test results showed that the level of cell movement ability decreased significantly with the increase of the concentration of Juglone. Transwell invasion assay showed that Juglone had a strong inhibitory effect on the invasiveness of HeLa cells in vitro. Western blotting results showed that Juglone inhibited the expression of MMP-2 and MMP-9 protein in HeLa cells. Conclusion: Juglone can inhibit the invasion and metastasis in HeLa cells, and its possible mechanism may be related to down regulating the expression of MMP-2 and MMP-9.
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Objective To study the regulatory effect and molecular mechanism of juglone on apoptosis of cervical cancer Hela cells. Methods Cervical cancer Hela cells were cultured and treated with different dosages of juglone (10, 20, and 40 μmol/L, respectively) and c-Jun N-terminal kinase (JNK) inhibitor SP600125 (10, 20, and 40 μmol/L, respectively). Then cellular proliferative activity and the expression of JNK/c-Jun pathway molecule and apoptotic molecule in the cells were detected. Results After 6, 12, 18 and 24 h of treatment, the value for proliferative activity of cells treated with juglone was significantly lower than that of control group (P < 0.05), and the anti-proliferative effect was more significant as the treatment period and juglone dosage increased (P < 0.05). The protein expressions of Bax, CytC, Fas, FasL, Caspase-3, p-JNK and p-c-Jun in cells treated with juglone were significantly higher than those of control group (P < 0.05), and the protein expressions of Bax, CytC, Fas, FasL, Caspase-3, p-JNK and p-c-Jun increased more remarkably as the juglone dosage increased (P < 0.05). In cells treated with 40 μmol/L juglone and SP600125, the protein expressions of Bax, CytC, Fas, FasL and Caspase-3 were significantly lower than those of cells treated with 40 μmol/L juglone (P < 0.05), and the protein expressions of Bax, CytC, Fas, FasL and Caspase-3 reduced more remarkably as the SP600125 dosage increased (P < 0.05). Conclusion Juglone can increase the expression of apoptotic molecules in mitochondrial pathway and death receptor pathway by activating JNK/c-Jun pathway, thus inducing apoptosis of cervical cancer cells.
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OBJECTIVE@#To study the regulatory effect and molecular mechanism of juglone on apoptosis of cervical cancer Hela cells.@*METHODS@#Cervical cancer Hela cells were cultured and treated with different dosages of juglone (10, 20, and 40 μmol/L, respectively) and c-Jun N-terminal kinase (JNK) inhibitor SP600125 (10, 20, and 40 μmol/L, respectively). Then cellular proliferative activity and the expression of JNK/c-Jun pathway molecule and apoptotic molecule in the cells were detected.@*RESULTS@#After 6, 12, 18 and 24 h of treatment, the value for proliferative activity of cells treated with juglone was significantly lower than that of control group (P < 0.05), and the anti-proliferative effect was more significant as the treatment period and juglone dosage increased (P < 0.05). The protein expressions of Bax, CytC, Fas, FasL, Caspase-3, p-JNK and p-c-Jun in cells treated with juglone were significantly higher than those of control group (P < 0.05), and the protein expressions of Bax, CytC, Fas, FasL, Caspase-3, p-JNK and p-c-Jun increased more remarkably as the juglone dosage increased (P < 0.05). In cells treated with 40 μmol/L juglone and SP600125, the protein expressions of Bax, CytC, Fas, FasL and Caspase-3 were significantly lower than those of cells treated with 40 μmol/L juglone (P < 0.05), and the protein expressions of Bax, CytC, Fas, FasL and Caspase-3 reduced more remarkably as the SP600125 dosage increased (P < 0.05).@*CONCLUSION@#Juglone can increase the expression of apoptotic molecules in mitochondrial pathway and death receptor pathway by activating JNK/c-Jun pathway, thus inducing apoptosis of cervical cancer cells.
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OBJECTIVE: To investigate the effect and mechanism of juglone on the apoptosis of human gastric cancer SGC-7901 cells. METHODS: The antiproliferative effect of juglone on SGC-7901 cells was tested by the MTT assay. The apoptosis rate and in-tracellular reactive oxygen species(ROS) level were detected by flow cytometry (FCM). The expression of JNK, p-JNK, p38, and p-p38 proteins were examined by Western blot. In order to clarify the role of ROS in the apoptosis induced by juglone on SGC-7901 cells, the combination of the juglone and ROS inhibitor NAC groups were set up in each experiment above. RESULTS Juglone could effectively inhibit the proliferation of SGC-7901 cells (IC50 for 72 h was 24.16 μmol·L-1). When combination juglone with NAC, the IC50 raised to 36.91 μmol·L-1. After 72 h of exposure to 5-20 μmol·L-1 of juglone, the cell apoptosis rate increased gradually with the increase of juglone concentration. After adding NAC, the apoptosis rates declined and the apoptosis rate of 20 μmol·L-1 group decreased from 32.06% to 11.56%. After SGC-7901 cells were treated with juglone for 24 h, the ROS level increased and mitochondrial transmembrane potential decreased which were inhibited by the pretreatment of NAC. After 48 h of exposure to different con-centration of juglone, the expressions of p-p38 and p-JNK proteins were up-regulated which could also be inhibited by the adding of NAC. Meanwhile, there were no significant changes in p38 and JNK protein expression in all groups. CONCLUSION: Juglone can induce apoptosis of human gastric cancer SGC-7901 cells by JNK and P38 pathway mediated by reactive oxygen species.
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Objective:To explore the mechanism of promotion effect of juglone combined with cisplatin on the apoptosis of human cervical cancer HeLa cells,and to clarify the effects of its associated signal transduction pathways.Methods:The HeLa cells at logarithmic growth phase were divided into control group,juglone group, cisplatin group and juglone combined with cisplatin group (combined treatment group).The inhibitory rates of proliferation of HeLa cells were detected by MTT assay.The apoptosis was detected by Hoechst 33258 staining. The expressions of Bcl-2, Bax and caspase-3, AKt, and pAKt were detected by Western blotting method. Results:The MTT results showed that the HeLa cell proliferation at 24,48 72 h in each drug group was inhibited;compared with control group,the profileration of HeLa cells in juglone group and cisplatin group was significantly inhibited,especially in combined group. Compared with single drug group,the inhibitory effect in combined treatment group was more significantly.After treatment for 12 h,the typical morphological changes of apoptosis were found in juglone group and cisplatin group by Hoechst 33258 staining,especially in combined treatment group. The Western blotting results showed that the expression levels of Bcl-2 and pAKt in HeLa cells in juglone group and cisplatin group 12 h after treatment were decreased obviously,whereas the expression levels of Bax,Caspase-3,and AKt were increased significantly, especially in combined treatment group compared with control group. Conclusion:Juglone combined with cisplatin could inhibit the PI3K/AKt pathway,thereby promoting the apoptpsis of HeLa cells.
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OBJECTIVE:To study the effects of different drying methods on the quality of North Qinglongyi,thus optimize the best drying way of North Qinglongyi. METHODS:North Qinglongyi was processed using five kinds of drying methods includ-ing drying in the shade,drying in the sun,oven drying,microwave drying,freeze drying and oven drying under different tempera-ture (40,50,60 ℃). The indicators were investigated,such as drying time,recovery rate of water,water content,alcohol ex-tract,content of active ingredients walnut quinone and juglone,in vitro anti-tumor activity of methanol extract to human gastric ade-nocarcinoma BGC823 cells and human lung cancer A549 cells. The effects of different drying methods on the quality of North Qinglongyi were analyzed comparatively. RESULTS:Among 5 kinds of drying method,microwave drying time was the shortest, and the time of drying in the shade was the longest;water content of sample with freeze drying was the highest,and that with mi-crowave drying was the lowest;the content of alcohol extract has little difference;the content of walnut quinone of sample with freeze drying was the highest,and that with microwave drying was the lowest;the content of juglone of sample dried at 40 ℃ was the highest,and that of freeze drying was the lowest;anti-tumor experiment in vitro showed that inhibition rate of sample with dif-ferent drying methods on the growth of 2 kinds of cells was in descending order of freeze drying>40 ℃ drying>drying in the shade,and the inhibition rate decreased with the increase of drying temperature. CONCLUSIONS:Different drying methods have obvious influence on the quality of North Qinglongyi. 40 ℃ drying method is the best for North Qinglongyi drying from the com-prehensive analysis including cost,the contents of effective composition,anti-tumor activity and practice.
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OBJECTIVE To explore the pro-apoptotic mechanism of juglone in SiHa cells and to in?vestigate its associated signal transduction pathways. METHODS SiHa cells were treated with juglone 20μmol·L-1 for 24,48 and 72 h. Cellular morphology was detected by inverted microscopy.The cell viability was detected by methyl thiazolyl tetrazolium (MTT) assay. After 24 h treatment with juglone 20μmol·L-1,the cell apoptosis was detected by flow cytometry while the expressions of apopto?sis-related protein BCL-2,BAX and cleave-caspase-3,PI3K/AKt pathway-related protein PTEN,AKT and pAKT were detected by Western blotting. RESULTS After treatment with juglone for 24, 48 and 72 h,the growth of SiHa cells was significantly inhibited. Compared with cell control group,cells in juglone treated gruop were sparse,slipped off the wall,became round and the cell proliferation inhibitory rate was 43.3%,63.0%and 73.1%(P<0.05,P<0.01),respectively. Twenty-four hours post treatment, the early apoptosis rate of juglone treated gruop cells was increased by(6.47±1.79)%(P<0.01)compared with cell control group. Western blotting results showed that the expression of BCL-2 decreased by 53.0%while the expression of BAX and caspase-3 increased by 85.5%and 183.3%,respectively. The expression of PTEN was increased by 75.0% but the pAKt was decreased by 45.8%(P<0.01). CONCLUSION Juglone can upregulate the expression of PTEN, thus inhibiting PI3K/AKt pathway and promoting apoptosis of SiHa cells.
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Objective To investigate the effects and mechanisms of pin1(peptidyl-prolyl isomerase 1) inhibitor ju?glone on migration and invasion in glioma cell line U-87 MG. Methods Glioma cells were treated with juglone at 0, 0.8, 1.6 and 3.2μmol·L-1. Wound-healing assay and invasion assay were performed to examine the inhibitory activity of ju?glone on glioma cell line U-87 MG. Western blot was used to analyze the protein expression of β-catenin, VEGF, MMP-2 and MMP-9. Results The wound-healing assay showed that the wound-healing rate in juglone-treated groups was 46.04%±6.25%and 30.05%±13.35%at concentrations of 1.6μmol·L-1 and 3.2μmol·L-1 , repectively. Juglone treat?ment significantly reduced the wound-healing rate compared with controls (P<0.05). Transwell invasion assay showed that the number of invaded cells in juglone–treated groups was 103.67 ± 5.69 and 77.33 ± 7.77 at the concentrations of 1.6μmol·L-1 and 3.2μmol·L-1 , respectively. Juglone treatment significantly reduced cell invasion compared with con?trols (P<0.05). Treatment with juglone significantly down-regulated the expression levels ofβ-catenin, VEGF, MMP-2 and MMP-9 in U-87 MG cells in a dose-dependent manner. Conclusion The present data suggests that juglone has a significant inhibitory action on cell migration and invasion through down-regulation of theβ-catenin and its downstream VEGF, MMP-2 and MMP-9 protein expressions in glioma cell line U-87 MG.
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AIM:To explore the effect of peptidyl-prolyl cis/trans isomerase (Pin1) inhibitor juglone on apop-tosis of human cervical cancer SiHa cells.METHODS:Cultured SiHa cells were incubated with juglone at concentrations of 10, 20, 50, 80 and 100 μmol/L for 24 h.The SiHa cell activity was detected by methyl thiazolyl tetrazolium ( MTT) assay.The cell apoptosis was analyzed by flow cytometry with Hoechst 33258 staining.The protein levels of cleaved caspase-3,8,9 and PTEN was determined by Western blotting.RESULTS:In different doses of juglone groups, the SiHa cell growth was greatly inhibited ( P<0.05) in a dose-dependent manner as compared with control group.The IC50 of ju-glone was 20.4 μmol/L.After treatment with juglone at concentration of 20 μmol/L for 12 h, the apoptosis of SiHa cells was induced, and the typical morphological changes of cell apoptosis such as karyopyknotic pyknic hyperfluorescence bolus, nuclear fragmentation and apoptotic body were observed by Hoechst 33258 staining.The early apoptotic rate was increased significantly as compared with the control.The protein levels of cleaved caspase-3, 8, 9 and PTEN were also increased sig-nificantly as compared with control group.CONCLUSION:Juglone significantly inhibits the cell activity and induces the apoptosis of SiHa cells in vitro by inhibiting the caspase pathway and increasing the expression of anti-oncogene.
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Objective To study the effect of different concentrations of juglone on the proliferation and apoptosis of human cervical cancer cells (HeLa), and explore the anti-tumor effect of juglone in vitro. Methods HeLa cells were cultured in vitro for 24 h and then divided into control group and juglone-treated group. The cells in juglone-treated group were cultured again with 10, 20, 50, 100 and 200μmol/L juglone, respectively, for 24h. The morphological changes in HeLa cells were observed with inverted microscope. The proliferation of HeLa cells was assessed detected by methyl thiazolyl tetrazolium (MTT) assay. The cell cycle and apoptosis were observed by flow cytometry (FCM). The expression of caspase-3 in HeLa cells was determined by caspase-3 colorimetric assay kit. Results Under morphological observation, it was found that different concentrations of juglone led to change in cell shape. MTT assay showed that juglone significantly inhibited the growth of HeLa cells in a dose-dependent manner (P<0.05 or P<0.01). FCM assay indicated that the percentage of the cell cycles arresting at G2/M phase after treatment with above concentrations of juglone for 24 h were increased to 12.92%, 16.23%, 23.64%, 34.22% and 52.64% from 7.5%, respectively (P<0.05 or P<0.01). The caspase-3 activity in juglone-treated HeLa cells remarkably increased in a dose-dependent manner as compared with control group. Conclusion Various concentrations of juglone can inhibit the proliferation and induce the apoptosis of HeLa cells in a dose-dependent manner, implying that juglone may have anti-tumor effect.