表皮生长因子对PC-3细胞内皮素-1及其受体mRNA表达的影响

目的：探讨表皮生长因子（EGF）对激素非依赖性前列腺癌（HRPC）PC-3细胞中内皮素1（ET-1）及其受体mRNA表达的影响。方法：EGF作用不同时间（0、8、16、24、32、48h）后，RT-PCR法测定PC-3细胞中ET-1及其受体ETBR mRNA、ETBR mRNA表达；EGF干预24h后，RT-PCR法测定ET-1及其受体ETAR mRNA、ETBR mRNA表达变化。结果：在PC-3细胞中可检测到ET-1及ETAR mRNA表达，但无ETBR mRNA表达；EGF可上调ET-1及ETAR mRNA表达，与对照组比较，差异具有显著性；ET-1及ETAR mRNA表达随EGF干预时间增加而增加，EGF作用不同时间对PC-3细胞ET-1、ETAR mRNA表达的影响不同，差异具有显著性（P〈0．05）。结论：EGF可上调PC-3细胞中ET-1及ETAR mRNA表达，为HRPC的治疗提供了分子生物学基础。     

高糖对人腹膜间皮细胞局部内皮素系统表达的影响

目的:研究高浓度葡萄糖对人腹膜间皮细胞(HPMC)局部内皮素系统表达的影响,并应用非特异性内皮素受体拮抗剂即ETA/ETBR拮抗剂PD142893进行干预,探讨高浓度葡萄糖透析液导致腹膜受损的机制.方法:以人腹膜间皮细胞株(HMrSV5)为研究对象,采用放免法检测不同时间(12 h、24 h和48 h)、不同浓度(50、100、150、200、250 mmol/L)的葡萄糖、甘露醇作用下HPMC分泌的ET-1水平.采用半定量逆转录多聚酶链反应(RT-PCR)检测局部内皮素系统ET-1、ETAR、ETBR及ECE的基因表达.采用ELISA检测不同浓度ET-1、葡萄糖作用下IL-1β水平,并在此基础上加入PD142893,观察IL-1β的变化.结果:(1)正常HPMC分泌低水平的ET-1,高浓度的葡萄糖可诱导HPMC分泌 ET-1增加,呈时间和剂量依赖.高浓度的甘露醇作用下,HPMC分泌的ET-1与正常对照组相比未见明显差异.(2)正常HPMC表达完整的ET-1、ETAR、ETBR、ECE mRNA.高糖作用下ET-1、ETAR、ETBR mRNA表达增强,而高糖对ECE mRNA的表达无明显影响.(3)ET-1、高糖刺激HPMC分泌IL-1β,呈剂量依赖.PD142893可明显减少高糖诱导的IL-1β的分泌.结论:正常HPMC存在局部内皮素系统,高浓度葡萄糖诱导其表达增加,高浓度葡萄糖透析液导致腹膜受损可能部分通过内皮素途径,内皮素可能是导致CAPD患者腹膜纤维化的重要因素.     

内皮素受体拮抗剂对前列腺癌细胞PC3和成骨细胞相互作用的影响

目的 在细胞共培养微环境下,观察内皮素(ET)受体拮抗剂阻断内皮素轴对前列腺癌细胞PC3和成骨细胞SaOS_2相互作用的影响.方法 利用细胞体外共培养系统,比较前列腺癌细胞PC3和成骨细胞SaOS_2在共培养微环境下和单独培养环境下细胞增殖的差异.共培养微环境下,分别以ET_A受体(ET_AR)拮抗剂BQ123和ET_B受体(ET_BR)拮抗剂BQ788阻断相应的ET受体,观察其对前列腺癌细胞和成骨细胞增殖的影响.同时,通过酶联免疫吸附试验(ELISA)法测定各组培养液中的ET-I浓度,通过逆转录.聚合酶链反应(RT-PCR)测定PC3细胞、SaOS_2:细胞ET-I及其受体的mRNA表达,比较它们的表达差异.结果 与单独培养比较,共培养微环境下的PC3细胞和SaOS_2细胞的增殖数量均显著升高(P均＜0.05).ET_AR拮抗剂BQ123能阻断共培养微环境对SeOS:细胞的生长促进作用,部分阻断共培养微环境对PC3细胞的生长促进作用,而ETBR拮抗剂BQ788则无显著的干预作用.EUSA结果显示共培养组培养液中ET-1浓度[(14.26±1.06)βg/L/10~5个细胞]显著高于单独培养组[(6.58±0.74)pc/L/105个细胞,P＜0.05].RT-PCR结果显示PC3细胞表达ET-1和ET_AR,SaOS_2细胞只表达ET_AR;与单独培养比较,共培养微环境下PC3细胞ET-l的表达显著升高(P＜0.05),而PC3细胞和SaOS2细胞ET_AR的表达差异均无统计学意义(P＞0.05).结论 ET-1及ET_AR构成的内皮素轴是共培养微环境中前列腺癌细胞PC3和成骨细胞之间相互作用促进增殖的重要链接因子,而ETAR拮抗剂能有效地抑制两者间的相互作用,抑制PC3细胞增殖.     

内皮素-1及其受体在良性前列腺增生中的表达研究

目的探讨内皮素-1及其受体(ETAR与ETBR)在良性前列腺增生(BPH)移行区组织中表达的意义。方法采用免疫组化及Western blot技术检测5例正常前列腺组织(NP)与16例BPH组织中内皮素-1及其受体表达情况。结果BPH移行区组织中内皮素-1阳性面积为(77936.16±85291.33)μm2,ETAR积分吸光度为316.6±65.2,明显高于正常前列腺组织(阳性面积75.68±110.85μm2,ETAR积分吸光度为140.2±64.8),而2组ETBR的表达(积分吸光度分别为81.4±31.8,105.0±45.5)差异无统计学意义。结论BPH移行区组织中内皮素-1及其ETAR表达上调,在BPH的病理生理过程中可能产生重要作用。     

内皮素1及其受体拮抗剂对HSC-T6细胞内皮素受体mRNA表达的影响

目的探讨内皮素1(ET-1)及其受体拮抗剂对HSC-T6细胞内皮素受体mRNA表达的作用及其机制,进一步了解缩血管物质在门静脉高压症发病机制中的作用。方法培养的肝星状细胞HSC-T6细胞系分为7组,分别为空白对照组,ET-1组(培养瓶中加入10 nmol/L的ET-1),BQ-123组[加入1μmol/L的选择性内皮素A型受体(ETRA)拮抗剂BQ-123],BQ-788组[加入1μmol/L的选择性内皮素B型受体(ETRB)拮抗剂BQ-788],ET-1+BQ-123组(加入10 nmol/L的ET-1和1μmol/L的BQ-123),ET-1+BQ-788组(加入10 nmol/L的ET-1和1μmol/L的BQ-788),ET-1+BQ-123+BQ-788组(加入10 nmol/L的ET-1、1μmol/L的BQ-123和1μmol/L的BQ-788)。采用逆转录聚合酶链反应(RT-PCR)检测HSC-T6细胞内皮素受体mRNA的表达。结果ET-1+BQ123+BQ788组ETRA mRNA的表达与空白对照组相比差异具有统计学意义(分别为0.292±0.023和0.440±0.030,P<0.05),其余各组与之相比无明显差异(P>0.05);与ET-1组相比,ET-1+BQ788组和ET-1+BQ123+BQ788组ETRA mRNA表达低,差异具有统计学意义(分别为0.329±0.044,0.292±0.023和0.487±0.039,P<0.05];与空白对照组相比,ET-1组ETRB mRNA表达上调,但差异无统计学意义(分别为0.499±0.136和0.289±0.047,P=0.134];与ET-1组相比,ET-1+BQ788组ETRB mRNA表达明显低,差异具有统计学意义(分别为0.153±0.071和0.499±0.136,P<0.05)。结论ET-1对ETRA mRNA的表达无明显作用;ET-1本身可能会上调HSC-T6 ETRB mRNA的表达,ET-1作用于ETRA时则会抑制ETRB mRNA的表达。     

胆汁性肝硬化猪肝组织中内皮素受体及诱导型一氧化氮合酶mRNA表达的意义

目的　研究胆汁性肝硬化猪肝组织中内皮素受体 (ETAR、ETBR)及诱导型一氧化氮合酶 (iNOS)mRNA的表达及其对肝脏血液循环的影响。方法　湖北白种猪 15头 ,其中实验组 10头 ,对照组 5头。监测门静脉压 (Pp)、门静脉血流量 (Qp)和平均动脉压 (MAP)的变化 ,用原位杂交方法检测肝组织中ETAR、ETBR及iNOSmRNA的表达水平。结果　术后 4周实验组Pp(14 .6±1.7)mmHg (1mmHg =0 .13 3kPa)、Qp(3 2 .3± 2 .8)ml·min-1·kg-1及MAP(80 .5± 2 .5 )mmHg显著高于对照组的 (8.1± 3 .6)mmHg、(14 .2± 3 .7)ml·min-1·kg-1和 (10 8.3± 6.5 )mmHg(P <0 .0 1)。实验组ETAR、ETBR及iNOSmRNA的表达水平分别为 0 .78± 0 .2 0、0 .90± 0 .2 3和 0 .75± 0 .13 ,均显著高于对照组 (分别为 0 .3 8± 0 .2 6、0 .2 4± 0 .15和 0 .3 2± 0 .11,P <0 .0 1)。结论　胆汁性肝硬化猪肝组织中ETAR、ETBR及iNOSmRNA表达水平的变化影响肝脏血液循环的调节。     

成纤维细胞生长因子受体1在胰腺癌细胞中的表达和调控

目的 研究成纤维细胞生长因子受体1(FGFR1)蛋白和mRNA在胰腺癌细胞系中的表达和调控机制.方法 采用Western免疫印迹实验、Northern印迹分析和RT-PCR检测FGFR1在胰腺癌细胞中的表达.使用外源性生长因子刺激细胞并使用激酶抑制剂阻断细胞内信号转导通路,观察FGFR1蛋白和mRNA在胰腺癌细胞中表达的变化情况.结果 FGFR1蛋白和mRNA在胰腺癌细胞系中均有不同程度的表达.生长因子刺激可上调FGFR1蛋白和mRNA的表达水平.其中IGF-1、EGF和FGF2显著增加Mia PaCa-2细胞FGFR1的表达,EGF和FGF2显著增加PANC-1细胞FGFR1的表达(P＜0.05).FGF2对FGFR1表达的调节具有时间依赖性.ERK1/2抑制剂UO126和p38 MAPK抑制剂SB203580降低了PANC-1细胞中FGFR1的蛋白和mRNA的表达水平. 结论生长因子可上调FGFR1在胰腺癌细胞中的表达水平,MAPK信号转导通路中的ERK1/2和p38 MAPK亚通路参与FGFR1表达的调节.     

PI-3K和p38MAPK通路在EGF诱导PC-3细胞环氧化酶-2表达上调中的作用

目的:研究p38丝裂原激活蛋白激酶(p38MAPK)和磷脂酰肌醇-3激酶(PI/3K)通路在表皮生长因子(EGF)诱导的激素非依赖性前列腺癌(hormone-refractory prostate cancer,HRPC)PC-3细胞环氧化酶2(cyclooxygen-ase-2,COX-2)表达上调中的作用。方法:MTT法检测EGF(0μg/L)、EGF(10μg/L)、EGF(10μg/L)+PI-3K阻断剂(LY294002,20μmol/L)、EGF(10μg/L)+p38MAPK阻断剂(SC203580,20μmol/L)处理后的细胞增殖情况。RT-PCR和Western印迹测定上述处理24h后PC-3细胞COX-2的表达变化,ELISA测定细胞培养液中前列腺素E2(PGE2)的变化。结果:LY294002和SC203580明显抑制EGF刺激后的PC-3细胞增殖(P<0.05)及EGF诱导的COX-2上调和PGE2生成(P<0.05)。结论:PI-3K通路和p38MAPK通路可能参与了EGF诱导的PC-3细胞COX-2的表达上调。     

内皮素受体在雄激素非依赖性前列腺癌中的作用

目的 研究雄激素非依赖性前列腺癌株PC3中内皮素(ET)受体表达及受体阻断后PC3细胞的凋亡情况。方法 RT-PCR法测定PC3中ET受体的表达以及采用受体拮抗剂干预后其表达的变化,通过流式细胞仪和透射电镜研究干预后PC3细胞凋亡的情况。结果 PC3中ETA表达较高,而ETB表达非常弱。ETA受体拮抗剂干预后,ETA表达明显减少,随干预浓度的增加,PC3细胞凋亡的比例逐渐增加;而ETB受体拮抗剂干预后无明显改变。结论 PC3中ET-1的作用主要通过ETA受体介导,ETB受体是静止的。ETA受体阻断后,表达减少,PC3细胞出现凋亡,并呈剂量依赖关系。     

脓毒症和脓毒性休克大鼠的肺肾肠内皮素-1和内皮素A受体基因表达变化及其意义

目的 观察大鼠脓毒症和脓毒性休克时内皮素-1(ET-1)和内皮素A受体(ETAR)基因在肺、肾和小肠黏膜中的表达变化以及各脏器功能受损情况.方法 24只雄性大鼠随机分为正常组、对照组、脓毒症组和脓毒性休克组;通过脓毒症和脓毒性休克大鼠模型,采用逆转录-聚合酶链反应(RT-PCR)以葡萄糖-6-磷酸脱氢酶(G-6-PD)基因为内参照基因,分别检测脓毒症和脓毒性休克组肾、肺和小肠黏膜组织ET-1和ETAR基因表达情况,同时检测其血清丙氨酸转氨酶(ALT)、白蛋白(A)、尿素氮(BUN)、肌酐(Cr)和动脉血气变化.结果 脓毒症组肾肺肠组织ET-1 mRNA表达相对量分别为87%、74%、82%,ETAR mRNA为82%、65%、78%;脓毒性休克组肾肺肠组织ET-1 mRNA表达相对量分别为98%、85%、93%,ETAR mRNA为95%、80%、91%,较正常组和对照组均明显增加(P<0.01).脓毒性休克组大鼠血清ALT为288.50±194.10、BUN 58.00±5.20,Cr1 33.67±18.70,较正常组、对照组和脓毒症组均显著上升,差异有统计学意义(P<0.01);脓毒症组血清ALT为97.17±6.30、BUN 30.17±2.30、Cr 75.83±6.70,较正常组明显升高(P<0.01).脓毒症组和脓毒性休克组早期动脉血气分析为低氧和呼吸性碱中毒,脓毒性休克组后期出现明显的低氧血症和CO2潴留.结论 ET-1和ETAR参与了脓毒症和脓毒性休克的病理生理过程,且可能是参与脓毒症和脓毒性休克并发多脏器功能障碍综合征(MODS)启动的因子之一.     

Endothelin-1 production by prostate cancer cell lines is up-regulated by factors involved in cancer progression and down-regulated by androgens.

BACKGROUND: Recent data demonstrate that endothelin-1 (ET-1) concentration increases in plasma of men with advanced, hormone-refractory prostate adenocarcinoma. In addition, ET-1 is involved in osteblastic remodelling and new bone formation, suggesting a role for this vasoactive peptide in the metastatic progression of prostate cancer to the bone. METHODS: We investigated the regulation of ET-1 expression in androgen-sensitive and insensitive prostate cancer cell lines by androgens and several factors involved in progression of prostate cancer (EGF) and bone remodelling (TGFbeta-1, IL1-alpha and IGF-1). RESULTS: Northern analysis and radio immunoassay demonstrated that all the ET-1 pathways are tuned off in the androgen-sensitive LNCaP cell line when compared to the androgen-insensitive PC-3 and DU145. In PC-3 cells transfected with a full-length androgen receptor expression vector (PC-3-AR), treatment with androgens reduced gene expression and secretion of ET-1 without affecting the gene expression of ET-3. Collectively, these data support a role for androgens in the regulation of ET-1 production by prostate adenocarcinoma cells. In PC-3 and DU145 cells, ET-1 gene expression and secretion were up-regulated by TGFbeta-1, EGF and IL1-alpha, whereas IGF-1 was ineffective. Conversely, none of the treatments affected ECE-1 or ET-3 gene expression. CONCLUSIONS: In conclusion, ET-1 production by prostate adenocarcinoma cells is down-regulated by androgens and up-regulated by factors involved in tumour progression indicating a role for this peptide in the biology of prostate cancer. In view of the role exerted by ET-1 in the process of bone metastasis, our data suggest the use of ET-1 receptor antagonists in the treatment of advanced prostate cancer.     

前列腺癌与良性前列腺增生细胞株差异核基质蛋白的鉴定

目的:鉴定人前列腺癌雄激素依赖性细胞株LNCaP、前列腺癌雄激素非依赖性细胞株PC-3与良性前列腺增生细胞株BPH-1之间差异表达的核基质蛋白(nuclear matrix proteins,NMPs)。方法:常规制备各细胞株NMPs,应用双向凝胶电泳对其进行分离;对差异表达的蛋白质点行MALDI-TOF-MS/MS质谱分析,数据库搜索并鉴定。结果:成功获得了分辨率高、重复性好的不同细胞株NMPs双向凝胶电泳图谱;初步鉴定出包含酶、调节蛋白、RNA结合蛋白及各种因子在内的12个差异表达的蛋白质,在癌细胞株中3个NMPs表达上调,9个下调。结论:人前列腺癌细胞与良性前列腺增生细胞之间NMPs表达存在明显差异;初步筛选出12个差异NMPs,其表达水平及功能与疾病的联系需进一步研究。     

Prostasin基因在前列腺细胞株中表达情况研究

目的 探讨Prostasin在前列腺癌中的功能。方法 运用RT-PCR方法检测人前列腺增生细胞株BPH-1，无转移能力前列腺癌细胞株LNCaP，及有转移能力的前列腺癌细胞株PC-3、DU-145中Prostasin基闪表达情况。结果 Prostasin基斟在BPH-l和LNCaP中正常表达，在PC-3、DU-145中低表达。BPH-l中Prostasin的表达与DU-145和PC-3比较差异有显著性，同样LNCaP中Prostasin的表达与DU-145和PC-3比较差异亦有显著性（P〈0．01）。BPH-1与LNCaP之间表达差异无显著性，DU-145和PC-3之间表达差异亦无显著性，（P〉0.05）。结论 ProStasin可能是前列腺癌的转移抑制剂。     

前列腺癌细胞系及组织中α-1,6岩藻糖基转移酶的表达及意义

目的 探讨α-1,6岩藻糖基转移酶(α-1,6 fucocyltransferase,FUT8)在不同前列腺癌细胞系、癌及癌旁组织中的表达差异及其意义.方法 培养前列腺正常上皮细胞(RWPE-1)、4种前列腺癌细胞(22RV1、LNCap、PC-3、DU145)及2种其他类型泌尿系统肿瘤细胞(T-24、786-O),收集15对前列腺癌及癌旁组织,提取总RNA,采用RT-PCR检测不同细胞系、癌及癌旁组织中FUT8 mRNA的表达,并进行差异比较.结果 FUT8 mRNA在前列腺癌细胞系中的表达高于前列腺正常上皮细胞系(RWPE-1)、膀胱癌细胞系(T-24)以及肾癌细胞系(786-O).雄激素非依赖性前列腺癌细胞系(PC-3、DU145)中FUT8 mRNA的表达高于雄激素依赖性前列腺癌细胞系(LNCap);前列腺癌转移灶细胞系(PC-3、DU145)中FUT8 mRNA的表达高于前列腺癌原发灶细胞系(22RV1).同时还发现,FUT8 mRNA在前列腺癌组织中的表达高于癌旁组织.以上差异均有统计学意义(P<0.01).结论 FUT8在不同前列腺癌细胞系、癌及癌旁组织中存在差异表达,可能参与前列腺癌的雄激素非依赖性转化及进展转移等过程.     

NF-κB抑制剂对前列腺癌PC-3细胞STAT3核转位的影响

目的:观察IL-6刺激后的前列腺癌PC-3细胞STAT3和NF-κB的表达情况;验证NF-κB抑制剂咖啡酸苯乙基酯(CAPE)对PC-3细胞IL-6和STAT3表达的影响。方法:20 ng/ml IL-6分别作用于PC-3细胞0、5、10、20、30、45 min后,Western印迹和实时荧光定量PCR检测STAT3和NF-κB蛋白和mRNA水平的表达差异;流式细胞技术检测细胞周期。采用TNF-α或TNF-α联合CAPE作用于PC-3细胞,收集培养液上清,ELISA检测IL-6的表达;同时用Western印迹检测p-STAT3的表达。结果:IL-6刺激PC-3细胞后,p-STAT3蛋白的表达明显上调,细胞增殖指数明显增高。TNF-α作用于PC-3细胞后,培养液中IL-6的表达上调,同时p-STAT3蛋白的表达亦上调(P<0.05)。CAPE联合TNF-α作用于PC-3细胞后,培养液中IL-6的表达及p-STAT3蛋白的表达均明显低于TNF-α作用后的表达水平(P<0.05)。结论:CAPE能抑制TNF-α引起的IL-6的分泌,从而抑制IL-6引起的STAT3核转位;通过CAPE抑制NF-κB表达,继而影响STAT3等相关细胞信号传导途径,可能成为前列腺癌治疗的一条新途径。     

Epidermal growth factor receptor mRNA levels in human prostatic tumors and cell lines.

The mitogenic activity of epidermal growth factor (EGF) is mediated by a cell surface receptor (EGF-R) which has been identified in human prostate tissues. Because of conflicting reports on the relative levels of EGF-R in prostate tumors as measured by binding of radiolabelled EGF, we have examined EGF-R expression at the level of the specific messenger RNA using a sensitive RNase protection assay. Expression of the mRNA for EGF-R was higher in carcinoma (CaP, N = 38) than in benign prostatic hyperplasia (BPH, N = 35) samples (p less than 0.01). The highest levels of EGF-R mRNA were found in the human prostatic carcinoma cell lines, PC-3 and DU145. Among the CaP samples, there was an association of higher EGF-R mRNA levels with higher tumor extent and dedifferentiation. Since EGF has also been found in prostatic tissues, the enhanced expression of the EGF-R gene may play a role in the growth of prostate tumors, possibly by an autocrine pathway.     

Prostate cancer cell proliferation is influenced by leptin

BACKGROUND: Obesity is considered a risk for many cancers. Serum leptin levels are often elevated in obese people. Leptin acts as a mitogenic agent in many tissues; therefore, it may act to promote cancer cell growth. We previously demonstrated that leptin acts as a growth factor for prostate cancer cells in vitro. The purpose of this study was to characterize leptin receptor isoform mRNA expression in leptin-treated DU145 and PC-3 prostate cancer cell lines. Expression levels of SOCS-3, a known leptin-inducible suppressor of leptin signaling, and known mitogenic signaling pathways of PI3K and ERK were also analyzed METHODS: DU145 and PC-3 cells were treated with 0, 4, 40, or 80 ng/ml leptin for 0, 0.5, 1, 2, 4, 24, or 48 h. Multiplex RT-PCR was performed to determine mRNA levels of the short (huOB-Ra) or the long (huOB-Rb) OB-R isoforms or SOCS-3. p-Akt and p-ERK were determined by Western blot. Cell viability and apoptosis were determined by MTT and nucleosomal fragmentation assay RESULTS: DU145 and PC-3 expressed huOB-Ra, huOB-Rb, and SOCS-3 mRNA. huOB-Ra mRNA levels increased in PC-3 at 48 h (P < 0.01); however, no significant changes were observed in DU145. huOB-Rb mRNA levels decreased at 48 h in DU145; however, a twofold increase at 48 h (P < 0.01) was observed with PC-3 and was dose-dependent (P < 0.05). Leptin increased SOCS-3 mRNA in DU145 at 24 and 48 h (P < 0.05) and in PC-3 at 1 h (2-fold) and 48 h (fivefold; P < 0.01). Leptin up-regulated p-Akt in a time- and dose-dependent manner in the DU145 prostate cancer cells via a suppression of apoptosis. Leptin up-regulated p-ERK in a time-dependent manner in PC-3 cells CONCLUSIONS: In prostate cancer cells, the mitogenic effects of leptin are not a consequence of altered receptor isoform mRNA expression. No defect in SOCS-3 signaling was observed, and proliferation appears to be working through the PI3K and MAPK leptin receptor-activated pathways, depending on cell type. Leptin stimulation may be selective for either pathway to suppress apoptosis, thereby enhancing prostate cancer growth.     

环氧化酶2在不同前列腺癌细胞系中的表达

目的:研究环氧化酶2(COX-2)在不同前列腺癌细胞系中的表达,探讨COX-2在前列腺癌侵袭进展及转移潜能获得机制中的可能作用。方法:应用Western印迹及RT-PCR鉴定LNCaP及其亚细胞系C4-2和AR-CaP亚细胞系IF11、IA8,以及PC-3细胞中COX-2的表达情况,并初步分析其在不同特性前列腺癌细胞系转移侵袭过程中的作用。结果:Western印迹结果显示:COX-2蛋白在PC-3细胞中表达相对较高,在IF11、IA8、LNCaP和C4-2细胞中表达缺失,差异具有统计学意义(P<0.05)。COX-2mRNA表达结果同蛋白一致。结论:不同来源、不同转移潜能的前列腺癌细胞株中COX-2表达存在差异。高表达COX-2可能在PC-3细胞高侵袭转移潜能获得方面起着一定作用,而与其他细胞系转移作用无关。     

The role of survivin and Bcl-2 in zinc-induced apoptosis in prostate cancer cells

ObjectivesTo study the effects of zinc treatment on the gene expression levels of survivin and Bcl-2 in prostate cancer cells.Materials and methodsThe effects of zinc exposure on apoptosis were assessed using two human prostate cancer cell lines, LNCaP and PC-3. Zinc-induced apoptosis was measured by Annexin V staining. The direct effect of zinc on the expression levels of zinc transporters (ZnT-1 and ZnT-4) and apoptosis-related genes (Bax, Bcl-2, and survivin) was determined by RT-PCR analysis.ResultsWhen LNCaP and PC-3 cells were exposed to various concentrations of zinc sulfate for 48 hors, their growth was inhibited in a dose-dependent manner. The levels of zinc in both cell lines treated with zinc sulfate for 24 hours were higher than in untreated cells. Exposure to zinc induced apoptosis and necrosis in LNCaP and PC-3 cells. Apoptosis became more extensive as the treatment time with zinc increased. There was a significant increase in the gene expression levels of ZnT-1 and ZnT-4 in both cell lines treated with zinc sulfate compared with untreated cells. The expression of Bax mRNA was up-regulated, while the expression of Bcl-2 and survivin were decreased in both cell lines following zinc treatment.ConclusionsExposure to zinc sulfate in human prostate cancer cells increased intracellular levels of zinc, which resulted in increased apoptosis. The apoptogenic effect of elevated concentration of zinc could be due either to increased expression of zinc transporters and increased levels of Bax or decreased Bcl-2 and survivin expression.