The multikinase inhibitor sorafenib induces caspase-dependent apoptosis in PC-3 prostate cancer cells

The present study investigated the effects of the multikinase inhibitor sorafenib on androgen-independent can- cer cells viability and intracellular signaling. Human androgen-independent PC-3 prostate cancer cells were treated with sorafenib. At concentration that suppresses extracellular signal-regulated kinase phosphorylation, sorafenib treatment reduced the mitochondrial transmembrane potential. Sorafenib also down-modulated the levels of mye- loid cell leukemia 1, survivin and cellular inhibitor of apoptosis protein 2. Sorafenib induced caspase-3 cleavage and the mitochondrial release of cytochrome c. However, no nuclear translocation of apoptosis inducing factor was detected after treatment and the pan-caspase inhibitor Z-VAD-FMK had an obvious protective effect against the drug. In conclusion, sorafenib induces apoptosis through a caspase-dependent mechanism with down-regulated antiapoptotic proteins in androgen-independent prostate cancer cells in vitro.

Inhibitory effect of a new gossypol derivative apogossypolone (ApoG2) on xenograft of human prostate cancer cell line PC-3

Objective: To investigate the inhibitory effect of apogossypolone (ApoG2) on prostate cancer cell line PC-3 in vivo, and explore its mechanism. Methods: The models of transplantation tumors in Balb/c nu/nu mice were established via subcutaneous injection of PC-3 cells and the tumor-transplanted mice were divided into 4 groups: control group and three ApoG2 treatment groups, with 10 mice in each group. Volumes of the tumor were estimated every 2 d and the morphology of tumor tissues was observed. Immunohistochemistry was employed to observe the expression of Bcl-2, PCNA, CD31, caspase-3 and caspase-8 in tumor tissues. Results: ApoG2 (2.5 mg/kg-10 mg/kg) given intraperitoneally once a day can obviously inhibit the growth of subcutaneous prostatic carcinoma implant. The tumor volume decreased obviously when the treatment dosage was bigger than 5.0 mg/kg (P<0.01). Meanwhile, ApoG2 decreased the expression of PCNA and CD31, and enhanced the expression of caspases-3, caspase-8 in tumor tissues. Conclusion: ApoG2 exert an inhibitory effect on prostatic carcinoma possibly by inducing apoptosis and inhibiting tumor angiogenesis.

Tumor Necrosis Factor-related Apoptosis Ligand Induces Apoptosis in Prostate Cancer PC-3M Cell Line

Summary： To study the effect of tumor necrosis factor-related apoptosis inducing ligand （TRAIL） on PC-3M cell line, PC-3M cell line was incubated with gradient concentrations of TRAIL for 4-24 h. Annixin-V fluorescence staining and TUNEL method were employed to detect the apoptosis of PC-3M cells. The morphology of apoptotic PC-3M cells was observed by electron microscopy. The relationship between TRAIL concentrations and the percentage of apoptotic cells was evaluated by flow cytometry. The proliferation inhibitory ratio was calculated by using MTT colorimetry. Our results showed that apoptosis of PC-3M cells could be induced by treatment with TRAIL for at most 4 h. The results of flow cytometry and MTT colorimetry demonstrated a time-and concentration-dependent relationship between cell apoptosis rate and TRAIL concentration. It is concluded that apoptosis of PC-3M cells can be induced by TRAIL. Because of the selective killing effect of TRAIL on tumor cells, it may become a potential alternative for the treatment of advanced prostate cancer.

Effect of hypoxia-inducible factor-1α on proliferation and invasion of prostate cancer PC-3 cell in hypoxic situation

Objective We transfected recombinant expression plasmid of pcDNA3. 1-HIF-1α into prostate cancer cells, to research effect of HIF-1α on proliferation of prostate cancer cell PC-3. Methods We selected a stable expression cell line with G418 we selected by transfection

Contragestazol (DL111-IT) inhibits proliferation of human androgen-independent prostate cancer cell line PC3 in vitro and in vivo

Aim： To evaluate the antiproliferative activity of contragestazol （DL111-IT） on the human prostate cancer cell line PC3 in vitro and in vivo and to elucidate its potential molecular mechanisms. Methods： The cell killing ability of DL111-IT was measured by the 3-（4,5-dimethylthia-zol,2-yl）-2,5-diphenyltetrazolium bromide （MTT） reagent assay method and the tumor xenograft model. The cell cycle was analyzed by flow cytometry and protein expression, including retinoblastoma （pRb）, cyclin-dependent kinase 4 （CDK4） and cyclin D 1, was detected by Western blotting. Results： DL111-IT exhibited high efficiency on cell growth inhibition of the human androgen-independent prostate cancer cell line PC3. The drug concentration that yielded 50 % cell inhibition （IC50 value） was 9.9 mg/mL. In the PC3 tumor xenograft study, DL111-IT （1.25 mg/kg-20.0 mg/kg） given once a day for 10 days significantly inhibited tumor growth, with the inhibition rate ranging from 21% to 50 %. Flow cytometric analysis indicated that DL111-IT could cause GI arrest in the PC3 cell line, but not apoptosis. DL111-IT enhanced pRb expression and down-regulated CDK4 and cyclin D 1 expression, suggesting that cell cycle regulation might contribute to the anticancer property of DL 111- IT. Conclusion： DL111-1T inhibits the proliferation of human androgen-independent prostate cancer cell line PC3 in vitro and in vivo by a cell cycle regulation pathway.

The effect of interleukin 6 on the growth of LNCaP and PC-3 prostatic carcinoma cells

Objective To investigate the effect of IL-6 on prostatic carcinoma cell lines, and differential effects on androgen-dependent and androgen-independent prostatic carcinoma cells. Methods The IL-6 producing capacities of LNCaP and PC-3 cells were determined, and effects of exogenous IL-6 and anti-IL - 6 antibodies on LNCaP and PC - 3 cells were examined. Results LNCaP produced a very small amount of IL-6, but PC-3 produced more, the concentraion of IL-6 being 190 pg/48 h per ml(1 × 106). The exogenous IL-6 inhibited LNCaP growth significantly,but had no obvious effect on PC -3 cells. Anti-IL-6 antibodies lowered PC-3 cells growth rate but had neutral effect on LNCaP. Conclusion PC-3 cells produces IL-6 massively in autocrine manner. IL-6 could be antagonized by anti-IL-6 antibodies,resulting in slowing PC-3 cells growth, and LNCaP cells growth could be inhibited by exogenous IL-6.7 refs,2 tabs.

Emodin induces apoptosis in human prostate cancer cell LNCaP

Aim： To elucidate effects and mechanisms of emodin in prostate cancer cells. Methods： Viability of emodin-treated LNCaP cells and PC-3 cells was measured by MTT assay. Following emodin treatments, DNA fragmentation was assayed by agarose gel electrophoresis. Apoptosis rate and the expression of Fas and FasL were assayed by flow cytometric analysis. The mRNA expression levels of androgen receptor （AR）, prostate-specific antigen （PSA）, p53, p21, Bcl-2, Bax, caspase-3, -8, -9 and Fas were detected by RT-PCR, and the protein expression levels of AR, p53 and p21 were detected by Western blot analysis. Results： In contrast to PC-3, emodin caused a marked increase in apoptosis and a decrease in cell proliferation in LNCaP cells. The expression of AR and PSA was decreased and the expression of p53 and p21 was increased as the emodin concentrations were increased. In the same time, emodin induced apoptosis of LNCaP cells through the upregulation of caspase-3 and -9, as well as the increase of Bax/Bcl-2 ratio. However, it did not involve modulation of Fas or caspase-8 protein expression. Conclusion： In prostate cancer cell line, LNCaP, emodin inhibites the proliferation by AR and p53-p21 pathways, and induces apoptosis via the mitochondrial pathway.

Manganese antagonizes iron blocking mitochondrial aconitase expression in human prostate carcinoma cells

Aim： To investigate the possible role of manganese in the regulation of mitochondrial aconitase （mACON） activity human prostate carcinoma cell line PC-3 cells. Methods： The mACON enzymatic activities of human prostate carcinoma cell line PC-3 cells were determined using a reduced nicotinamide adenine dinucleotide-coupled assay. Immunoblot and transient gene expression assays were used to study gene expression of the mACON. The putative response element for gene expression was identified using reporter assays with site-directed mutagenesis and electrophoretic mobility-shift assays. Results： In vitro study revealed that manganese chloride （MnCI2） treatment for 16 h inhibited the enzymatic activity of mACON, which induced the inhibition of citrate utility and cell proliferation of PC- 3 cells. Although results from transient gene expression assays showed that MnCI2 treatment upregulated gene translation by approximately 5-fold through the iron response element pathway, immunoblot and reporter assays showed that MnCl2 treatments inhibited protein and gene expression of mACON. This effect was reversed by cotreatment with ferric ammonium citrate. Additional reporter assays with site-directed mutagenesis and electrophoretic mobility-shift assays suggested that a putative metal response element in the promoter of the mACON gene was involved in the regulation of MnCh on the gene expression of mACON. Conclusion： These findings suggest that manganese acts as an antagonist of iron, disrupting the enzymatic activity and gene expression of mACON and citrate metabolism in the prostate.

Manganese antagonizes iron blocking mitochondrial aconitase expression in human prostate carcinoma cells

Aim:To investigate the possible role of manganese in the regulation of mitochondrial aconitase(mACON)activity human prostate carcinoma cell line PC-3 cells.Methods:The mACON enzymatic activities of human prostate carcinoma cell line PC-3 cells were determined using a reduced nicotinamide adenine dmucleotide-coupled assay. Immunoblot and transient gene expression assays were used to study gene expression of the mACON.The putative response element for gene expression was identified using reporter assays with site-directed mutagenesis and electro- phoretic mobility-shift assays.Results:In vitro study revealed that manganese chloride(MnCl2)treatment for 16h inhibited the enzymatic activity of mACON,which induced the inhibition of citrate utility and cell proliferation of PC- 3 cells.Although results from transient gene expression assays showed that MnCl_2,treatment upregulated gene translation by approximately 5-fold through the iron response element pathway,immunoblot and reporter assays showed that MnCl_2 treatments inhibited protein and gene expression of mACON.This effect was reversed by co- treatment with fenic ammonium citrate.Additional reporter assays with site-directed mutagenesis and electrophoretic mobility-shift assays suggested that a putative metal response element in the promoter of the mACON gene was involved in the regulation of MnCl_2 on the gene expression of mACON.Conclusion:These findings suggest that manganese acts as an antagonist of iron,disrupting the enzymatic activity and gene expression of mACON and citrate metabolism in the prostate.

双氢青蒿素对前列腺癌PC-3细胞自噬的诱导作用及其机制

目的探讨双氢青蒿素(DHA)对前列腺癌PC-3细胞的自噬诱导作用及其机制。方法用0、12.5、25、50、100μmol/L DHA处理PC-3细胞,采用CCK-8法检测细胞活力,克隆形成实验检测细胞增殖能力。取PC-3细胞,设置对照组(不做处理)、DHA组(50μmol/L DHA处理48h)、自噬抑制剂3-MA组(5mmol/L 3-MA处理48h)、DHA+3-MA组(50μmol/L DHA+5mmol/L 3-MA处理48h),采用Western blotting和RT-qPCR检测自噬相关蛋白[微管相关蛋白轻链3B(LC3B)、酵母Atg6同源物(Beclin-1)]的表达情况,透射电镜观察自噬小体形成情况,使用自噬双标慢病毒mCherry-GFP-LC3B转染PC-3细胞检测自噬流变化,CCK-8法检测细胞活力,流式细胞术检测细胞凋亡率。设置对照组(不做处理)、DHA组(50μmol/L DHA处理48h)、ROS抑制剂NAC组(5mmol/L NAC处理48h)、DHA+NAC组(50μmol/L DHA+5mmol/L NAC处理48h),采用Western blotting检测ROS/AMPK/mTOR信号通路相关蛋白的表达。用50μmol/L DHA处理PC-3细胞48h后提取总蛋白,分成Input组(全蛋白裂解液)、IP组(加入Beclin-1抗体)、IgG组(加入同等质量的IgG),采用免疫共沉淀(Co-IP)实验检测Beclin-1与Vps34、Bcl-2及HMGB1蛋白的相互作用。结果CCK-8法检测结果显示,PC-3细胞存活率随着DHA浓度的升高而降低,且呈剂量和时间依赖性(P<0.05);DHA作用24、48、72h的半数抑制浓度(IC50)分别为97.12、57.10、29.35μmol/L,据此选择50μmol/L DHA作用48h进行后续实验。克隆形成实验结果显示,PC-3细胞克隆形成率随着DHA浓度的升高而明显降低(P<0.01)。Western blotting和RT-qPCR检测结果显示,与对照组比较,DHA组PC-3细胞中Beclin-1、LC3B mRNA和蛋白表达水平明显升高(P<0.01);与DHA组比较,DHA+3-MA组PC-3细胞中Beclin-1、LC3B mRNA和蛋白表达水平明显降低(P<0.01)。透射电镜观察可见DHA组PC-3细胞中出现明显的自噬小体,且自噬小体数较对照组明显增多(P<0.05)。mCherry-GFP-LC3B慢病毒转染实验结果显示,与对照组比较,DHA组每细胞红黄斑点比增高(P<0.01),DHA+3-MA组每细胞红黄斑点比降低(P<0.01)。与DHA组比较,DHA+3-MA组细胞存活率降低,凋亡率增高(P<0.01)。与对照组比较,DHA组PC-3细胞中p-mTOR蛋白相对表达水平降低(P<0.05),p-AMPK蛋白相对表达水平升高(P<0.01);与DHA组比较,DHA+NAC组p-mTOR蛋白相对表达水平升高(P<0.01),p-AMPK蛋白相对表达水平降低(P<0.01)。Co-IP实验结果显示,DHA处理后Beclin-1与Bcl-2的作用减弱,与Vps34、HMGB1的结合增强。结论DHA可诱导前列腺癌PC-3细胞发生自噬,其机制可能与调控自噬相关基因Beclin-1、LC3及ROS/AMPK/mTOR信号通路有关。

Therapeutic effect of oridonin on mice with prostate cancer

Objective: To investigate the therapeutic effect and the related mechanism of oridonin on mice with prostate cancer. Methods: Sixty BALB/C male nude mice were selected. A model of RM-1 cell transplantation tumor of prostate cancer was built by the subcutaneous inoculation of RM-1 cells. After that, those 60 experimental mice were randomly divided into groups A, B and C. Each group had 20 mice. Mice in group A were treated with 0.2 m L of normal saline(0.9%) by intraperitoneal injection once a day; mice in group B received intraperitoneal injection of 1.875 mg/m L of oridonin once a day; and mice in group C received intraperitoneal injection of 7.5 mg/m L of oridonin once a day. Mice in the three groups were treated uninterruptedly for 5 weeks and were all killed. Then, tumors were excised and weighed to calculate their growth inhibitory rate, volume increment and anti-tumor rate. Thymus and spleen of mice in the three groups were collected to calculate the thymus and spleen index. Immunohistochemical staining was applied to observe the expression of caspase-3 in prostate cancer tissue of mice of the three groups. Results: The qualities and volume increment of tumors in groups B and C were significantly lower than those of group A(P < 0.05); the qualities and volume increment of tumors in groups C were evidently lower than those of group B(P < 0.05); the tumor volume increment and anti-tumor rate in group C were obviously higher than those of group B(P < 0.05); the thymus and spleen indexes of groups B and C were distinctly higher than those of group A(P < 0.05); comparison of the thymus and spleen indexes between group B and group C showed no statistical differences(P > 0.05). Immumohistochemical staining revealed that the caspase-3 protein in prostate cancer tissue of mice of group A expressed negatively with colourless or light-colored karyon; while the caspase-3 protein in prostate cancer tissue of mice of group B expressed positively with dark-colored karyon, centralized distribution and granular sensation; and the caspase-3 in prostate cancer tissue of mice of group C showed strong positive expression with big and darker colored karyon and dense distribution. Conclusions: Oridonin can inhibit the growth of RM-1 prostate cancer cells effectively and have great therapeutic effects on RM-1 cell transplantation tumor of prostate cancer.

KHSRP通过ANK3调节前列腺癌细胞对雄激素的反应性

目的·研究KH型剪接调节蛋白(KH-type splicing regulatory protein,KHSRP)对前列腺癌细胞增殖能力的影响及其下游基因的表达调控,考察KHSRP在前列腺癌由雄激素依赖型向非依赖型转变过程中的潜在作用及机制。方法·通过重组慢病毒感染雄激素依赖型前列腺癌LNCaP细胞和雄激素非依赖型前列腺癌DU145细胞,分别构建KHSRP功能性缺失/获得的稳定细胞株,并比较KHSRP在2种细胞之间的功能差异。采用蛋白质印迹法(Western blotting)检测稳定细胞株中KHSRP、雄激素受体(androgen receptor,AR)及锚定蛋白3(ankyrin 3,ANK3)的表达量。通过细胞增殖实验、平板克隆实验、小鼠成瘤实验等检测KHSRP对LNCaP细胞增殖能力的影响。通过转录组测序(RNA sequencing,RNA-seq)检测KHSRP影响的下游基因,并通过实时荧光定量PCR(quantitative real-time PCR,RT-qPCR)检测KHSRP下游基因的mRNA表达量。结合癌细胞系百科全书(Cancer Cell Line Encyclopedia,CCLE)、癌症基因组图谱(The Cancer Genome Atlas,TCGA)、基因表达综合(Gene Expression Omnibus,GEO)数据库,分析研究ANK3和KHSRP在前列腺组织样本中的表达情况以及ANK3在不同前列腺癌细胞株中的表达差异。结果·GEO数据分析结果显示KHSRP在前列腺癌组织中的表达高于良性前列腺组织,提示其与前列腺癌的发生相关。细胞增殖实验、平板克隆实验、小鼠成瘤实验结果显示KHSRP的表达与LNCaP细胞增殖能力呈负相关,提示KHSRP可以抑制前列腺癌细胞的增殖,且KHSRP对LNCaP细胞增殖能力的影响强于对DU145细胞的影响。对LNCaP稳定细胞株的Western blotting和RT-qPCR检测显示KSHRP过表达后LNCaP细胞中AR蛋白质水平下降,但mRNA水平未产生变化,提示KHSRP可以间接调节AR的蛋白质水平。RNA-seq和RT-qPCR结果显示KHSRP与影响AR蛋白稳定性的调节因子ANK3的表达呈正相关,随后的Western blotting证明KHSRP过表达后ANK3蛋白质的表达量明显升高,TCGA数据分析结果进一步说明KHSRP与ANK3的mRNA表达的相关性。根据CCLE和GEO数据,ANK3的表达与前列腺癌的恶性程度也密切相关。结论·KHSRP可能通过ANK3间接调控AR的蛋白稳定性,影响雄激素依赖型前列腺癌细胞的增殖能力,并介导前列腺癌细胞对雄激素反应性的改变。

组织DDX5、PBX3的表达对前列腺癌术后复发的预测价值

目的探讨与分析组织死亡盒蛋白5(Dead-boxpolypeptide5,DDX5)、前B细胞白血病同源盒基因3(pre-B-cell leukemia homeobox 3,PBX3)的表达对前列腺癌术后复发的预测价值。方法采用回顾性方法,2018年8月到2022年5月选择在南阳市第一人民医院诊治的前列腺癌患者90例作为研究对象,采用免疫组化法检测癌组织DDX5、PBX3表达水平。随访患者的术后复发情况并进行预测价值分析。结果90例患者随访到2022年8月,平均随访时间为16.20±2.22个月,复发10例(复发组),占比11.1%,其中吻合口复发2例,淋巴结复发9例。复发组的DDX5、PBX3表达阳性率为80.0%、90.0%,显著高于非复发组的26.3%、21.3%(P<0.05)。Spearsman分析显示DDX5、PBX3表达阳性率与术后复发存在相关性(P<0.05);COX回归分析显示DDX5、PBX3表达阳性率为影响术后复发的重要因素(P<0.05);ROC曲线显示DDX5、PBX3表达阳性率预测术后复发的曲线下面积分别为0.806、0.857。结论前列腺癌术后复发比较常见,多伴随有DDX5、PBX3的高表达,组织DDX5、PBX3的表达对前列腺癌术后复发的预测具有重要价值。

槲皮素对前列腺癌PC-3细胞凋亡作用的研究

目的:研究槲皮素对人前列腺癌PC-3细胞的凋亡作用。方法:体外培养PC-3细胞,给予不同浓度(50、100、150、200、250μmol/L)的槲皮素处理,MTT法检测槲皮素对细胞抑制率;流式细胞仪检测细胞凋亡率;透射电镜观察细胞超微结构的变化。结果:槲皮素能抑制人前列腺癌PC-3细胞的体外生长,且呈时间与剂量依赖性,浓度由低到高,其24 h的抑制率(%)分别为3.01±1.32、4.84±1.73、20.35±1.30、16.78±1.89、27.25±4.01,48 h的抑制率(%)分别为10.18±1.16、6.22±0.04、24.29±4.19、22.4±4.26、41.42±5.43,当药物浓度>150μmol/L时P<0.05,具有统计学意义;流式细胞术显示PC-3细胞凋亡率随槲皮素浓度升高和作用时间延长而增加(P<0.05),其中浓度150μmol/L和200μmol/L组,24 h的凋亡率(%)分别19.10±0.28、26.55±0.78,48 h的凋亡率(%)分别为27.65±1.06、38.30±5.96;电镜观察到细胞的典型凋亡形态学变化。结论:在适当的条件下,槲皮素对人前列腺癌PC-3细胞增殖有明显的抑制作用,同时可诱导PC-3细胞凋亡,其具体作用机制有待进一步深入研究。

沙棘黄酮制备及体外抑制人前列腺癌PC-3细胞作用研究

采用AB-8大孔树脂纯化沙棘黄酮初提物,用10%、30%、50%乙醇溶液梯度洗脱,以总黄酮含量和黄酮苷元含量为指标,筛选纯化样品;采用MTT法、实时无标记细胞分析技术检测沙棘黄酮样品体外对人前列腺癌PC-3细胞的增殖抑制作用,流式细胞技术检测细胞凋亡和细胞周期情况,Western blot检测Bax和Bcl-2蛋白表达情况。实验结果表明50%乙醇梯度洗脱样品(S50)中,总黄酮含量达到25.42%;水解后得到SH50样品,其槲皮素、异鼠李素含量分别达到5.03%、18.64%;25μg/m L的SH50样品对PC-3细胞即有增殖抑制作用,且呈剂量和时间依赖性;同时25μg/m L的SH50样品作用48 h即可显著诱导细胞凋亡(P<0.01),并将细胞周期阻滞在G2/M期,此外,样品可提高Bax蛋白和降低Bcl-2蛋白的表达。说明沙棘黄酮SH50样品对体外人前列腺癌PC-3细胞具有抑制增殖并诱导细胞凋亡作用,其机制可能与阻滞细胞周期,调节Bax和Bcl-2蛋白的表达有关。

参附注射液对前列腺癌PC-3细胞凋亡诱导的影响

目的:研究参附注射液(SF)对人前列腺癌PC-3细胞凋亡的影响及可能机制。方法:实验设立对照组和SF 50、100、200μl/ml组,Annexin V/PI染色流式细胞术检测细胞凋亡,RT-PCR检测p53 mRNA表达。结果:与对照组比较,作用24、48、72 h后,SF 50、100、200μl/ml组PC-3细胞存活率显著减少(P均<0.05)。SF各组24 h存活率分别为(93.76±2.63)%、(81.21±1.80)%、(18.01±3.84)%;48 h存活率分别为(94.67±1.11)%、(78.33±2.89)%、(10.34±1.44)%;72 h存活率分别为(91.30±0.47)%、(36.67±1.56)%、(1.33±0.32)%,呈浓度和时间依赖性。作用48 h后,p53 mRNA表达明显升高(P<0.05)。结论:SF可以诱导PC-3细胞凋亡,其作用机制可能与p53表达增高相关。

雷帕霉素对人前列腺癌PC-3细胞的作用及其机制

背景与目的:哺乳动物细胞中雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)信号途径调节细胞生长、增殖、存活和凋亡。本实验是研究雷帕霉素对前列腺癌PC-3细胞的作用及其机制。方法:培养前列腺癌PC-3细胞,采用MTT法检测雷帕霉素1nmol/L作用24h、36h、48h、72h后PC-3细胞增殖的改变,流式细胞术检测作用不同时间细胞周期的改变,Westernblot检测雷帕霉素作用PC-3细胞24h、36h、48h、72h后raptor、rictor、Akt、pS6k1-T389、pAkt-s473的表达情况。结果:MTT结果显示雷帕霉素在作用24h时,促进了PC-3细胞增殖,但在36h显著抑制PC-3细胞增殖(P<0.01),72h时抑制作用更明显。FCM结果显示雷帕霉素作用24h时S期细胞有所增加,但作用36、48、72h后PC-3细胞的G1期细胞逐渐增加,使PC-3细胞周期主要阻滞在G1期。Westernblot检测结果显示雷帕霉素作用24h时显著抑制raptor和pS6k1-T389的表达(P<0.01);rictor、Akt表达并没有显著变化;pAkt-s473表达在雷帕霉素作用24h时反而显著增加(P<0.01),但在36h后即被显著抑制,72h几乎完全被抑制(P<0.01)。结论:延长雷帕霉素作用时间可抑制PC-3细胞增殖,其机制可能与雷帕霉素阻滞细胞周期、抑制Akt磷酸化有关。

靛玉红对前列腺癌PC-3细胞增殖的抑制作用

目的:研究中药成分靛玉红对雄激素非依赖性前列腺癌PC-3细胞的杀伤作用及可能的机制。方法:采用MTT方法研究靛玉红对前列腺癌PC-3细胞增殖抑制作用,采用Western印迹检测细胞周期蛋白cyclin D1及c-myc表达,采用流式细胞术检测细胞周期。结果:靛玉红降低前列腺癌细胞的存活率呈浓度依赖性,当浓度为5μmol/L时,可降低存活率至52.2%,当浓度为10μmol/L时,PC-3细胞存活率降至13.6%。靛玉红浓度为5μmol/L时可明显抑制PC-3细胞的细胞周期,结果显示G0/G1期PC-3细胞增多,而与此同时S期和G2/M期细胞减少。此外,靛玉红可以抑制细胞周期进展关键调控蛋白cyclin D1,以及与之相关Wnt信号通路的下游基因c-myc的表达。结论:靛玉红可以抑制雄激素非依赖性前列腺癌PC-3细胞的增殖,其机制可能与其抑制细胞周期以及抑制Wnt信号通路有关。

昆布多糖硫酸酯对PC-3细胞PTEN和P27kip1基因表达的影响

目的:研究昆布多糖硫酸酯(LAMS)对体外培养的非激素依赖型人前列腺癌细胞株PC-3的作用,以及对PC-3细胞PTEN和P27kip1基因表达的影响,并探讨其抗肿瘤的作用机制。方法:分别用0、50、100、200μg/ml浓度的LAMS作用于PC-3细胞,用WST-8法检测其对细胞生长的抑制作用,利用流式细胞术(FCM)分析细胞周期和凋亡的变化,用荧光显微镜观察PC-3细胞形态的变化,RT-PCR和Western印迹检测细胞增殖、凋亡相关基因PTEN和P27kip1mRNA及蛋白的表达。结果:LAMS能抑制PC-3细胞的增殖,呈时间-剂量效应;不同浓度LAMS诱导PC-3细胞出现剂量依赖性S期阻滞;LAMS作用于PC-3细胞后凋亡率增加,并出现凋亡的形态学改变,相关基因PTEN和P27kip1mRNA及蛋白的表达呈剂量依赖性增加。结论:LAMS能抑制体外PC-3细胞的生长,并促进其S期阻滞和凋亡,PTEN和P27kip1mRNA及蛋白的表达明显增加,可能是其抑制前列腺癌生长的机制之一。

葡萄籽提取物对前列腺癌PC-3细胞的生长抑制作用

目的:观察葡萄籽提取物(GSE)对前列腺癌PC-3细胞的生长抑制作用。方法:经预试选用不同浓度(100、200、300μg/ml)的GSE分别作用PC-3细胞24、48、72h,以原代培养的1～3日龄SD大鼠肾细胞作为正常对照;采用四唑氮蓝(MTT)显色法检测GSE对PC-3细胞和SD大鼠肾细胞的生长抑制作用。结果:GSE以浓度和时间依赖性方式抑制PC-3细胞生长(P<0.01),对原代培养SD大鼠肾细胞生长仅有轻度抑制作用。结论:GSE可抑制前列腺癌PC-3细胞生长,有可能成为治疗前列腺癌的一种新药物。