ABSTRACT
OBJECTIVE：To study the effe cts of the water extract from Carpesium cernuum （AECC）on the proliferation ， metastasis and invasion of prostate cancer PC 3 cells. METHODS ：Cells were divided into control group and different concentration groups of AECC （5，10，20，40，80 μg/L），and then treated with relevant medicine or medium for different time （24，48，72 h）. The survival rates of cells were detected. Cells were divided into control group ，and AECC low ，medium and high concentration groups（20，40，80 μg/L）. After cultured for 24 h，Hoechst 33258 staining and flow cytometry were used to detect the apoptosis of cells. The number of cell metastasis and invasion were detected by Transwell assay. RT-qPCR and Western blot assay were applied to detect the mRNA and protein expression of β-catenin signaling pathway related migration and apoptosis proteins （β-catenin， MMP-7，c-Myc，caspase-3，Bcl-2 and Bax ）in AECC low and medium concentration groups. RESULTS ：With the increase of the concentration and culture time ，the survival rates of cells in AECC different concentration groups were significantly lower than control group （P＜0.05 or P＜0.01），and showed a decreasing trend. Compared with control group ，the early apoptosis rate （except the medium concentration group ）and the number of cell metastasis and invasion in AECC groups ，the mRNA and protein expression of MMP- 7，c-Myc（except for the low concentration group ）and Bcl- 2（except for mRNA of the low concentration group）in AECC low and medium concentration groups were decreased significantly （P＜0.05 or P＜0.01）. Late apoptosis rate of AECC groups ，the mRNA and protein expression of β-catenin，caspases-3（except for the low concentration group ），Bax（except for mRNA of the low concentration group ）in AECC low and medium concentration groups were increased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS ：AECC could inhibit the proliferation ，metastasis and invasion of PC 3 cells；the mechanism of which may be associated with regulating the expression of β-catenin signaling pathway related migration and apoptotic factors.
ABSTRACT
AIM:To explore the mitochondrial pathway in the apoptosis of PC3 cells induced by PXD101 (also named as belinostat).METHODS:PC3 cells were treated with PXD101 at different doses or stimulated with PXD101 for different time.The effect of PXD101 on the viability of PC3 cells was measured by CCK-8 assay.The apoptotic rates and the mitochondrial membrane potential (MMP) were analyzed by flow cytometry.The protein levels of Bcl-2, Bax and cytochrome C (Cyt C) were determined by Western blot.The caspase-3 activity were tested by caspase-3 activity assay kit.RESULTS:The viability of the PC3 cells was inhibited by PXD101 in a dose-and time-dependent manner.Flow cytometric analysis showed that the apoptotic rates were increased in the cells treated with PXD101 (P<0.01).At the same time, PXD101 induced the decreases in MMP and Bcl-2, the release of Cyt C, and the increase in caspase-3 activity.CONCLUSION:PXD101 induces the apoptosis of human prostate cancer cell line PC3 by mitochondrial signal pathway.
ABSTRACT
Objective To investigate the inhibitory effect of Coicis Semen oil on human prostate cancer PC-3 cell. Methods A Nude mice xenograft model was established with human prostate cancer PC-3 cells. Meanwhile, intragastric administration with Coicis Semen oil 2 mL/kg and 6 mL/kg was run for once daily and growth-curve was drawn through measuring the tumor volume until 26 d. At the end of the experiment, tumor weights were measured and the tumor inhibition rate of each group was calculated. The activity of fatty acid synthase (FAS) was determined through UV-VIS spectrophotometry. PC-3 cells were cultured in vitro. SYBR Green I real-time quantitative RT-PCR was used to determine the relative expression of FAS mRNA. Results The tumor inhibition rate of Coicis Semen oil 6 mL/kg group was 43.9%. Compared with model group, FAS activity in this tumor tissue decreased by 44.7% (P < 0.05). The expression of FAS mRNA was significantly decreased at 20 μL/mL of Coicis Semen oil group (P < 0.05). Conclusion Coicis Semen oil on human prostate cancer PC-3 cells has a significant inhibitory effect. FAS activity and mRNA expression decline may be related to its main anti-cancer mechanism.
ABSTRACT
Objective To investigate the effect of lentivirus carrying shRNA-VDR vector on GLi1 in pros-tate cancer PC-3 cells. Methods The cells were cultured according to the culture conditions of PC-3 cells. Expression of VDR and GLi1 in PC-3 cells was detected by fluorescence quantitative PCR and immunocytochemistry SP method.The efficiency of PC-3 cell virus infection was evaluated.The effect of VDR gene interference and GLi1 transcription level on PC-3 cells was detected by RT-PCR.Results Cell culture,cell status was recorded and PC-3 cells were in good condition and the passages was 4 days. Fluorescence quantitative and immunocytochemi-cal SP showed that VDR and GLi1 were expressed in PC-3 cells.The virus infection efficiency showed that the in-fection efficiency was about 95% when adding LV3-NC lentivirus to PC-3 cells at 1:10 ratio. RT-PCR showed that VDR-shRNA lentivirus successfully disturbed VDR expression and decreased by 85%(P < 0.05)compared with the control group after 72 days of VDR-shRNA lentivirus transfection. Transcription level of GLi1 gene in the experimental group increased by 9% compared with the control group(P < 0.05). The transcription level of GLi1 gene in the experimental group increased by 248% compared with the control group(P < 0.05). Conclusion The cultured PC-3 cells were in good condition. VDR and GLi1 genes were expressed in PC-3 cells. Lentivirus showed highest efficiency in infecting PC-3 at 1:10 ratio. When VDR was disturbed,the expression of GLi1 in-creased.In prostate cancer cells,vitamin D can inhibit the Hh signaling pathway and cause GLi1 expression down expression.
ABSTRACT
OBJECTIVE:To explore the effects of tamsulosin on proliferation and apoptosis in prostatic cancer PC-3 cells. METHODS:After treated with 0 (blank control group),12.5,25 and 50 μmol/L tamsulosin (tamsulosin low,medium and high-concentration groups)for 48 h,the viability of PC-3 cells was detected by MTT method. Hoechst 33258 staining was used to detect cell apoptosis rate. Western blot was used to determine the expression level of Bax and Bcl-2 protein,and the phosphoryla-tion level of protein kinase B(Akt),mammalian target rapamycin(mTOR),ribosomal S6 protein kinase(p70S6K)and 4E bind-ing protein 1(4E-BP1). RESULTS:Compared with blank control group,PC-3 cells viability and the phosphorylation level of Akt, p70S6K and 4E-BP1 decreased in tamsulosin low,medium and high-concentration groups,while expression level of Bax protein in-creased (P<0.05 or P<0.01);the apoptosis rate of PC-3 cells was increased in tamsulosin medium and high-concentration groups,while the expression level of Bcl-2 and phosphorylation level of mTOR were decreased(P<0.01),in concentration-depen-dent manner. CONCLUSIONS:Tamsulosin can inhibit PC-3 cells proliferation and induce cell apoptosis via blocking Akt/mTOR signal pathway.
ABSTRACT
Objective:To observe the effects of double-stranded small interfering RNA (siRNA) of the silent mating-type infor-mation regulation 2 homolog 1 (SIRT1) on the cell proliferation, cell cycle progression, and expression levels of the cell cycle negative regulators. These regulators include P21, P27, and phosphorylated retinoblastoma (PRb) proteins present in prostate cancer PC3 cells. This work further aims to explore the possible underlying mechanism for such effects. Methods:PC3 cells were cultured in vitro and then randomly divided into the mock group, scramble siRNA transfected group, and SIRT1 siRNA-transfected group. SIRT1 siRNA ef-ficiency was examined through reverse transcription polymerase chain reaction and Western blot analysis. The inhibitory rate of PC3 cell growth was determined through a methyl thiazolyl tetrazolium assay, and the cell cycle was investigated with the use of flow cytom-etry. The P21 and P27 protein expression levels and PRb status were determined by Western blot assay. Results:Compared with those of the mock and scramble siRNA groups, the expression levels of SIRT1 mRNA and protein significantly decreased in SIRT1 siR-NA-transfected cells. In addition, the inhibitory rate of PC3 cell growth was markedly increased, and the cell cycle of the PC3 cells was arrested at the G1 stage. The expression levels of negative cell cycle regulators, including P21 and P27 protein levels increased, whereas Rb protein phosphorylation was inhibited in SIRT1 siRNA-transfected PC3 cells. Conclusion: SIRT1 RNA interference inhibits PC3 cell growth and arrests cell cycle progression through the upregulation of the P21 and P27 proteins and the inhibition of Rb protein phosphorylation.
ABSTRACT
[ ABSTRACT] AIM: To study the autophagy of prostate cancer PC-3 cells induced by CD147 in vitro.ME-THODS:Themethod of amino acid starvation to induce autophagy was used.The expression of CD147 was detected by Western blotting.To study the functional effects of CD147 on autophagy in prostate cancer PC-3 cells, the down-regulation of CD147 expression was induced by the technique of RNAi.The conversion of autophagic marker protein LC3-I to LC3-II was determined by Western blotting.The cell death after starvation-induced autophagy was analyzed by trypan blue exclu-sion assay.RESULTS:The CD147 expression gradually increased in starvation-induced autophagy.The down-regulation of CD147 significantly increased the expression of autophagy-related protein LC3-II compared with control group.Mean-while, the cell death rates increased from (19.3 ±3.1)%and (22.3 ±3.5)%in control groups to (38.4 ±3.1)%in si-lencing the expression of CD147 in the PC-3 cells (P<0.05).CONCLUSION:CD147 inhibits starvation-induced auto-phgy and autophagy death in the prostate cancer PC-3 cells.
ABSTRACT
Objective To investigate the effect of gene silencing of Y-box binding protein-1(YB-1) by RNA interference on the proliferation and migration in prostate cancer cells lines PC-3 cells .Methods YB-1 siRNAs(pGenesil-1-YB-1-1 and pGenesil-1-YB-1-2) were synthesized and transfected into cloned into the the PC-3 cells by liposome .The expressions of YB-1 were measure by RT-PCR and Western blotting .The proliferation and migration were respectively detected by MTT and Transwell method . Results ThemRNA and protein expressions of YB-1 were significantly decreased by pGenesil-1-YB-1-1 and pGenesil-1-YB-1-2 (P<0 .05) ,compared with the control group ,the inhibition ratio of mRNA expression was 36 .23% and 39 .42% respectively and the inhibition ratio of protein expression was 41 .56% and 55 .33% respectively .The proliferation and migration were significantly decreased by pGenesil-1-YB-1-1 and pGenesil-1-YB-1-2(P<0 .05) .Conclusion YB-1 gene silencing by RNA interference inhibits the proliferation and migration in prostate cancer cells lines PC-3 cells .
ABSTRACT
Objective To study the anti-tumor effects of griseofulvin on human prostate cancer PC-3 cells both in vitro and in vivo. Methods PC-3 cells were treated with griseofulvin at various concentrations for48 h,cell survival rate was then measured by MTT assay.The changes of morphology were observed by fluorescence microscope; Annexin V-FITC apoptosis detection kit was used to detect apoptosis of the cells ; The enzyme activity changes of caspase-3,8,9 were detected by spectrophotometry.For in vivo study,we first established the PC-3 tumor model by grafting PC-3 cells in athymic nude mice,and then injected griseofulvin into the tumors.12 days after injection,the mice were sacrificed,the tumors were removed,weighed and the ratios of tumor-suppression were then calculated.We had detected the expressions of Bcl-2,Bax,p53 and Survivin with immumohistochemistry as well. Results MTT results showed that griseofulvin could significantly inhibit the proliferation of PC-3 cells in vitro in a dose-dependent manner,and the IC50 of griseofulvin was 18.17 ±2.10 μg/ml.Typical morphological changes of PC-3 cells were observed by microscope.The rates of apoptosis of griseofulvin treated PC-3 cells greatly increased compared with that of the control cells (31.37 ± 2.93％ vs 2.89 ± 0.67％,P ＜ 0.01 ).The caspase-3,caspase-8 and caspase-9 activities in griseofulvin treated PC-3 cells were significantly higher than those in control cells (0.562 ±0.050 vs0.113±0.014,0.337±0.053 vs 0.120±0.017,0.293±0.038 vs0.109±0.018,P＜0.01).On the 23th day after tumor vaccination,the tumor volume was 961.17 ± 78.12 mm3 in griseofulvin treated group and was 433.6 ± 12.8 mm3 in control group (P ＜ 0.01 ).The tumor weight was 742.50 ± 78.63 mg in griseofulvin treated group and was 1387.33 ± 71.47 mg3 in control group ( P ＜ 0.01 ).Bcl-2,Bax,p53 and Survivin protein expressions were 16.10 ± 3.45％,39.50 ± 6.88％,48.20 ± 8.04％,16.50 ± 2.22％ in griseofulvin treated group,respectively; 41.30 ± 3.95％,13.70 ± 2.98％,17.60 ± 3.21％,52.11 ± 6.28％ in control group,respectively.And there were significant differences in both groups (P ＜ 0.01 ).The in vivo data showed that griseofulvin suppressed the tumor growth conspicuously through down-regulating the expression of Bcl-2,Survivin,and up-regulating the expression of Bax,p53. Conclusions Griseofulvin can inhibit the growth of PC-3 and induce apoptosis of PC-3 cells.Griseofulvin inhibits the in vivo tumorigenicity of PC-3 cells.
ABSTRACT
Objective To investigate the effect of Huntingtin-interacting protein1 (HIP1) gene silencing on the proliferation of PC-3 cells in a human androgen-independent prostate tumor. Methods pSlience-shHIP1, a shRNA expression vector targeting HIP1, was constructed and transfected into PC-3 cells by liposome. RT-PCR was adopted to detect the silencing effect of HIP1 gene. Western bloting was then used to confirm the effective target point. Moreover, the scratch assay and growth curve assay were performed to analyze the effect of HIP1 on the proliferation of PC-3 cells. Results The efficiency of HIP1 gene silencing was increased to 83% (P<0.01) after PC-3 cell transfection. Western blotting proved that this genetic fragment was the effective target point for the inhibition of HIP1. The scratch assay and growth curve assay showed that the proliferation of PC-3 cells was inhibited by silencing HIP1 gene. Conclusions The pSilence-HIP1 expression vector can specifically suppress the expression of the HIP1 gene. In addition, HIP1 gene silencing can inhibit the proliferation and migration of PC-3 cells.
ABSTRACT
Objective To investigate the effect of VEGF ASODN in vitro and in vivo on the biological characteristics of human prostate cancer PC3 cells and its effect in xenotransplanted tumors in nude mice by local ASODN injection.MethodsVEGF ASODN was delivered into PC3 cells by Oligofectamine.There were three experimental groups: VEGF ASODN,VEGF ODN and control.Soft agar assay and matrigel invasion assay were used to measure cellular transformation and invasion ability,respectively.Tumor formation assay in nude mice was used to evaluate the effect of VEGF ASODN on proliferation of PC3 cells in vivo.The xenotransplanted prostate tumor model in nude mice was established and the effect of local ASODN injection on the inhibition of tumor growth in vivo was examed.ResultsThe soft agar colony numbers for control,ODN,and ASODN treated cells were 53.67±5.86,52.33±6.43 and 26.00±4.58,respectively (F =13.73,P＜0.01).The numbers of invaded cells for three group were 45.60±5.53,42.35±6.21 and 18.37±3.52,respectively (F =14.18,P ＜0.01).Tumor cells transfected with VEGF ASODN proliferated more slowly than other groups.28 days later after tumor cells were injected into nude mice,the tumor sizes of three groups were (1330.32±81.38) mm3,(1267.64±120.26) mm3 and (641.83±58.34) mm3 (F =17.26,P ＜0.01).After treating the transplanted tumor with VEGF ASODN or control oligos for four weeks,the tumor weight of three groups was (1.25±0.08) g,(1.17±0.06) g and (0.41±0.05) g,respectively.Comparing with control groups,the tumor inhibitory rates of ODN group and ASODN group were 6.4 ％ and 67.2 ％,respectively (x2=17.72,P＜0.005).Conclusion VEGF ASODN could inhibit VEGF expression in PC3 cells and lead to increasing cell apoptosis.After VEGF ASODN treatment,tumorigenesis in vitro is inhibited and cell invasion ability is decreased.The tumors originated from cells transfected with VEGF ASODN grow more slowly than control groups.Also local injection of VEGF ASODN could inhibit the growth of transplanted tumors in nude mice.
ABSTRACT
OBJECTIVES: Cancer has been investigated using various pre-targeting techniques or models focusing on radiobombesin analogues; however, both are not offered together. In this study, nano-bombesin labeling by a pre-targeting system was undertaken to develop an alternative approach for prostate tumor treatment. METHODS: A two-step pre-targeting system utilizing a combination of streptavidin (SA), biotinylated morpholino (B-MORF), biotinylated BBN (B-BBN) with two different spacers (b-Ala and PEG), and a radiolabeled cMORF was evaluated in vitro and in vivo. RESULTS: Final conjugation conditions consisted of a 1:1:2 ratio of SA:B-MORF:B-BBN, followed by addition of 99mTc-cMORF to compensate for free MORF. In vitro binding experiments with prostate cancer cells (PC-3) revealed that total binding was time-dependent for the Ala spacer but not for the PEG spacer. The highest accumulation (5.06 ± 1.98 percent) was achieved with 1 hour of incubation, decreasing as time progressed. Specific binding fell to 1.05 ± 0.35 percent. The pre-targeting biodistribution in healthy Swiss mice was measured at different time points, with the best responses observed for 7-h and 15-h incubations. The effector, 99mTc-MAG3-cMORF, was administered 2 h later. Strong kidney excretion was always documented. The greatest tumor uptake was 2.58 ± 0.59 percentID/g at 7 h for B-bAla-BBN, with a region of interest (ROI) value of 3.9 percent during imaging. The tumor/blood ratio was low due to the slow blood clearance; however, the tumor/muscle ratio was 5.95. CONCLUSIONS: The pre-targeting approach with a peptide was a viable concept. Further evaluation with modified sequences of MORF, including less cytosine, and additional test intervals could be worthwhile.
Subject(s)
Animals , Male , Mice , Bombesin/metabolism , Molecular Imaging/methods , Morpholines/pharmacokinetics , Nanoparticles , Prostatic Neoplasms/metabolism , Radioisotopes , Streptavidin/pharmacokinetics , Bombesin/analogs & derivatives , Bombesin , Cell Line, Tumor , Chromatography, High Pressure Liquid/methods , Disease Models, Animal , Mice, Nude , Organotechnetium Compounds , Prostatic Neoplasms , Random Allocation , Radioisotopes/chemistry , Time FactorsABSTRACT
We examined the effect of indole-3-carbinol (I3C, C9H9NO), an autolysis product of a glucosinolate and a glucobrassicin in vegetables, on MMP-2, -9 activities and TIMP-1 and -2 inductions via microtubule-associated protein kinase (MAPK) signaling pathway in prostate cancer cell line, PC3 cells. Our results indicated that I3C inhibited cell growth of PC3 cells in dose (0, 50, 100 micrometer) and time (0, 24, 48 and 72 h) dependent manners. Using gelatin zymography for MMP activity, we demonstrated that I3C significantly decrease MMP-2 and -9 activities in PC3 cells. We also observed that I3C decreased the proteins and mRNA levels of MMP-2 and -9 in PC3 cells as well. Inversely, expressions of TIMP-1 and -2 protein and mRNA in PC3 cells were increased by I3C in a dose dependent manner. In another experiment, we showed that I3C inhibited PC3 cells invasiveness by using marigel invasion assay and we also found that I3C suppressed MMP transcriptional activity by MAPK signaling pathways. Taken together, our results suggest that I3C may contribute to the potential beneficial food component to prevent the cancer metastasis in prostate cancer cells.
Subject(s)
Humans , Autolysis , Cell Line , Gelatin , Glucosinolates , Indoles , Neoplasm Metastasis , Prostate , Prostatic Neoplasms , Protein Kinases , Proteins , RNA, Messenger , Tissue Inhibitor of Metalloproteinase-1 , VegetablesABSTRACT
Objectives: To study the inhibitory effects of grape juice on human prostate carcinoma PC-3 cells and its possible mechanism. Method: PC-3 cells were treated with grape juice in different concentration(16, 32, 64 ?l/ml) and resveratrol for 48 h; and the proliferation of PC -3 cells were measured by growth curve and MTT assay. TUNEL was used to observe the morphology of apoptotic cells and flow cytometry to analyze the PC-3 apoptosis. Results: Resveratrol and grape juice could markedly inhibit the proliferation of PC-3 cells, and TUNEL positive cells were detectable. The percentages of apoptotic cells were 6.0%,18.5%,30.0%, 12.7% in experimental groups and resveratrol group respectively. The effect on PC-3 cells was enhanced with increasing amount of grape juice. The effect of low dose group was weaker than that of resveratrol group, but medium and high dose groups were stronger. Conclusion: The anti-prostate cancer effect of grape juice is not only related to resveratrol contained but also to synergism with other components, and its mechanism might be the induction of PC-3 cells apoptosis.