5-脂氧合酶抑制剂齐留通抑制二乙基亚硝胺诱导大鼠肝细胞癌形成的实验研究

目的：研究5-脂氧合酶（5-lipoxygenase,5-LOX）在原发性肝癌中的表达及其抑制剂齐留通对肝癌的影响和机制。方法：实验大鼠随机分为正常对照组（n=5）、模型组（n=8）及齐留通组（n=8）。HE染色观察肝癌病理图片,免疫组织化学染色检测5-LOX蛋白,M2型丙酮酸激酶（M2-PK）与细胞角蛋白19（CK19）的表达,RT-PCR检测肝组织5-LOX的表达,TUNEL检测肝癌细胞凋亡。结果：二乙基亚硝胺（DEN）可以诱导大鼠肝癌模型形成。模型组大鼠肝脏可见中等强度5-LOX的蛋白表达,较对照组明显增强（P〈0.01）,而齐留通组5-LOX的表达较模型组明显降低（P〈0.01）。免疫组织化学检测M2-PK在对照组阴性表达,模型组大鼠中等强度表达（P〈0.01）,齐留通组弱表达,较模型组差异显著（P〈0.05）。CK19在对照组大鼠中阴性表达,模型组大鼠较强表达（P〈0.01）,齐留通组中等表达,较模型组差异显著（P〈0.01）。对照组可检测出5-脂氧合酶mRNA的表达,但明显低于模型组,差异显著（P〈0.01）;齐留通组的表达介于二者之间,与模型组相比,有统计学意义（P〈0.01）。TUNEL法检测对照组几乎未见凋亡细胞,模型组大鼠肝脏仅见少数肝癌细胞阳性表达（30%-50%）,而齐留通治疗组大鼠肝脏见大量肝癌细胞阳性表达（70%-80%）,与模型组相比有统计学意义（P〈0.01）。结论：DEN诱发的大鼠原发性肝癌存在5-脂氧合酶表达,5-脂氧合酶抑制剂可防治DEN诱发的大鼠原发性肝癌的形成,其机制与诱导肝癌细胞凋亡相关。     

5-脂氧合酶抑制剂对人肝癌细胞株HepG2增殖和凋亡的影响

目的:研究5-脂氧合酶(5-lipoxygenase,5-Lox)在人肝癌细胞株HepG2中的表达,以及5-Lox抑制剂对人肝癌细胞株HepG2增殖、凋亡的影响。方法:应用免疫细胞化学染色及RT-PCR分别检测5-Lox的蛋白和mRNA在人肝癌细胞株HepG2中的表达,采用MTT法及流式细胞术检测5-Lox抑制剂齐留通(Zileuton)对人肝癌细胞株HepG2增殖、凋亡的影响。结果:5-Lox蛋白表达于肝癌细胞浆,局灶细胞核膜亦见阳性染色,RT-PCR可检测到肝癌细胞5-Lox的mRNA的表达。人肝癌细胞株HepG2给予不同浓度的齐留通处理24小时后,细胞存活率呈剂量依赖性下降;流式细胞术检测可见细胞凋亡率呈剂量依赖性升高。结论:人肝癌细胞株HepG2存在5-Lox的表达,5-Lox抑制剂齐留通可抑制人肝癌细胞株HepG2的增殖,诱导细胞凋亡。     

5-脂氧合酶抑制剂对人肝癌细胞株HepG2增殖和凋亡的影响

目的：研究5-脂氧合酶（5-lipoxygenase,5-Lox）在人肝癌细胞株HepG2中的表达,以及5-Lox抑制剂对人肝癌细胞株HepG2增殖、凋亡的影响。方法：应用免疫细胞化学染色及RT-PCR分别检测5-Lox的蛋白和mRNA在人肝癌细胞株HepG2中的表达,采用MTT法及流式细胞术检测5-Lox抑制剂齐留通（Zileuton）对人肝癌细胞株HepG2增殖、凋亡的影响。结果：5-Lox蛋白表达于肝癌细胞浆,局灶细胞核膜亦见阳性染色,RT-PCR可检测到肝癌细胞5-Lox的mRNA的表达。人肝癌细胞株HepG2给予不同浓度的齐留通处理24小时后,细胞存活率呈剂量依赖性下降;流式细胞术检测可见细胞凋亡率呈剂量依赖性升高。结论：人肝癌细胞株HepG2存在5-Lox的表达,5-Lox抑制剂齐留通可抑制人肝癌细胞株HepG2的增殖,诱导细胞凋亡。     

雷公藤内酯醇调节5-脂氧合酶对胰腺癌细胞增殖的影响

目的:探讨雷公藤内酯醇(Triptolide,TL)在胰腺癌细胞中5-脂氧合酶(5-LOX)蛋白的表达及对癌细胞生长的影响.方法:应用免疫细胞化学法检测胰腺癌细胞株SW1990和Capan2中5-LOX蛋白的表达,同时检测TL对胰腺癌细胞株SW1990 5-LOX蛋白表达的影响,采用MTT法和流式细胞术分别检测胰腺癌细胞SW1990的增殖和凋亡.结果:胰腺癌细胞株SW1990和Capan2中均表达5-LOX,并与细胞分化程度有关;TL抑制SW1990中5-LOX蛋白的表达,并抑制胰腺癌细胞SW1990增殖和促进细胞的凋亡,作用随浓度的增加而增强(P＜0.01);细胞的增殖和凋亡与5-LOX蛋白的表达相关(P＜0.01).结论:TL抑制胰腺癌细胞SW1990中5-LOX的表达,并抑制胰腺癌细胞增殖和诱导凋亡,5-LOX代谢途径在胰腺癌细胞的生长中发挥重要作用.     

5-脂氧合酶及Ki-67在星形细胞瘤中的表达及其相关性研究

目的:研究5-脂氧合酶(5-LOX)和Ki-67在星形细胞瘤中的表达及相关性.方法: 免疫组织化学法检测60例星形细胞瘤(低度恶性33例,高度恶性27例)中5-LOX和Ki-67的表达,逆转录聚合酶链反应(RT-PCR)方法检测5-LOX和Ki-67的mRNA水平,另取14例行脑外伤内减压术的脑组织作为对照.结果: 对照组中5-LOX和Ki-67不表达或低表达,星形细胞瘤中5-LOX阳性表达率66.7%(40/60),Ki-67阳性表达率71.7%(43/60);高度恶性组5-LOX和Ki-67 的阳性表达率高于低度恶性组(P<0.01);5-LOX和Ki-67表达呈正相关(γ＝0.875,P<0.01).星形细胞瘤5-LOX和Ki-67的mRNA含量高于正常脑组织(P<0.01),高度恶性组5-LOX和Ki-67的mRNA含量高于低度恶性组(P<0.01),5-LOX和Ki-67在星形细胞瘤中的mRNA水平呈正相关(γ＝0.781,P<0.01).结论: 5-LOX和Ki-67的表达与星形细胞瘤恶性程度有关,二者在星形细胞瘤发生发展过程中有协同作用.     

Smo、Gli1在大鼠肝癌中的表达及维生素D3对Smo、Gli1的抑制

背景与目的:研究表明Sonic Hedgehog(Shh)信号通路与肿瘤的发生有重要联系,本实验通过检测大鼠肝癌模型中Smo、Gli1的表达,探讨其与肝癌发生发展的关系,了解维生素D3(Vitamin D3,VitD3)对Smo、Gli1的作用.方法:取48只5～6周龄的健康雄性SD大鼠,并随机分组.对照组:一般饲养的8只大鼠;DEN组:二乙基亚硝胺(diethylnirtosamine,DEN)诱癌的20只大鼠;DEN+VitD3组:DEN+VitD3干预的20只大鼠;分别于实验12周和20周处死各组一半数量的大鼠,采用Western blot检测大鼠肝脏中Smo、Gli1蛋白的表达,实时荧光定量PCR检测两种蛋白mRNA的表达.结果:对照组中12周和20周大鼠肝脏的Smo、Gli1的mRNA及蛋白的表达差异无统计学意义(P＞0.05);DEN组或DEN+VitD3组中20周的Smo、Gli1表达高于12周(P＜0.05);相同实验周期:DEN的Smo、Gli1表达高于VitD3组(P＜0.05);与其他两组相比,对照组表达最低(P＜0.01).结论:Smo、Gli1可能参与肝癌的发生发展,VitD3对Smo、Gli1具有一定抑制作用.     

黄芩素诱导人肝癌细胞株SMMC-7721凋亡作用的研究

目的:研究12-脂氧合酶(12-lipoxygenase,12-LOX)在人肝癌细胞株SMMC-7721中的表达,以及选择性12-LOX抑制剂黄芩素对人肝癌细胞SMMC-7721生长、凋亡的影响.方法:采用免疫细胞化学染色及RT-PCR检测12-LOX的蛋白质和mRNA在SMMC-7721中的表达.应用MTT、流式细胞术及TUNEL检测细胞生长及凋亡.蛋白印迹法检测分析Caspase-3,Bcl-2,Bax,phospho-ERK1/2,ERK1/2和β-actin蛋白质的表达.结果:免疫细胞化学技术显示12-LOX蛋白表达于SMMC-7721细胞质.RT-PCR检测到黄芩素呈浓度-时间依赖性地抑制12-LOX mRNA表达.采用黄芩素小同浓度或不同时间段干预细胞SMMC-7721,MTT法检测细胞增殖呈剂量、时问依赖性的下降.原位末端转移酶标记技术及流式细胞仪分析证实12-LOX抑制剂诱导细胞凋亡.蛋白印迹法检测显示,黄芩素干预后,Bcl-2蛋白表达明显下降和Bax蛋白轻微上升,细胞凋亡蛋白酶-3活性水平与黄芩素浓度正相关,黄芩素对细胞凋亡相关蛋白的影响被12 (S)-HETE逆转,12(5)-HETE可能激活ERK1/2磷酸化.结论:人肝癌细胞株SMMC-7721存在12-LOX的表达,12-LOX抑制剂黄芩素可抑制细胞生长和诱导细胞凋亡.     

PDCD4对肝癌细胞生物学功能的影响机制及其在化疗敏感性中的作用

目的 探讨程序性细胞死亡-4(PDCD4)对肝癌细胞生物学功能的影响机制及其在化疗敏感性中的作用.方法 应用qRT-PCR和Western Blot检测肝癌细胞HepG2、SMMC-7721、MHCC97H和正常的肝细胞L02中PDCD4的水平.采用肝癌细胞HepG2为研究对象,将转染PDCD4过表达载体(pDsRed2-N1-PDCD4)的HepG2细胞设置为PDCD4过表达组;将60μmol/L替莫唑胺作用24 h的HepG2细胞设置为替莫唑胺组;将正常肝癌细胞HepG2(不加替莫唑胺)设置为对照组;将转染pDsRed2-N1-PDCD4载体后经60μmol/L替莫唑胺作用24 h的HepG2细胞设置为联合组.采用噻唑蓝(MTT)法检测各组细胞的存活率,流式细胞仪检测细胞凋亡情况,West-ern Blot检测细胞中Bax、Bcl-2、MMP-9表达水平.结果 PDCD4在肝癌细胞HepG2、SMMC-7721、MHCC97H中的表达水平均低于正常的肝细胞L02(P<0.01),肝癌细胞HepG2中的PDCD4表达水平最低;PDCD4过表达组、替莫唑胺组、联合组的肝癌细胞存活率均明显低于对照组(P<0.01);PDCD4过表达组、替莫唑胺组、联合组的肝癌细胞凋亡率均明显高于对照组(P<0.01);PDCD4过表达组、替莫唑胺组、联合组肝癌细胞中Bax的表达水平均明显高于对照组(P<0.01);Bcl-2、MMP-9的表达水平均明显低于对照组(P<0.01);联合组肝癌细胞中Bax的表达水平明显高于PDCD4过表达组和替莫唑胺组(P<0.01).结论 PDCD4在肝癌细胞中低表达,PDCD4可抑制肝癌细胞的增殖,促进肝癌细胞的凋亡,增加肝癌细胞的化疗敏感性,其作用机制可能与Bax、Bcl-2、MMP-9有关.     

苦参碱和氧化苦参碱对二乙基亚硝胺诱发大鼠肝癌的预防阻断作用

目的 :研究苦参碱 (MT)和氧化苦参碱 (OMT)分别对二乙基亚硝胺 (DEN)诱发大鼠肝癌的预防阻断作用。方法 :采用 0 .0 1%的DEN诱发大鼠肝癌 90d ,同时分别腹腔注射MT和OMT注射液 15mg/kg ,停止诱癌及给药处理 30d后 ,处死大鼠 ,观察大鼠肝脏的病理改变、肝表面癌结节数、肝 /体重比和血清中丙氨酸氨基转移酶 (ALT)、γ 谷氨酰转肽酶 (γ GT)、碱性磷酸酶 (ALP)的变化。结果 :MT组和OMT组大鼠的体重明显高于模型组 ,肝表面癌结节数、肝 /体重比和血清ALT、γ GT明显低于模型组 (P<0 0 5) ,而ALP较模型组有所升高。另外 ,OMT组的肝重、肝 /体重比和血清ALP均明显低于MT组(P <0 0 5)。结论 :MT和OMT ,尤其是OMT ,尽管不能完全阻断DEN诱发大鼠肝癌的发生 ,但能保护肝细胞免受损伤 ,延缓DEN诱发大鼠肝癌的形成     

TUM2-PK在肝癌中的表达及临床意义

目的:研究肿瘤M2型丙酮酸激酶(TUM2-PK)在肝癌中的表达及其与肿瘤临床病理特征的关系。方法:RT-PCR法检测42例肝细胞肝癌组织及相应癌旁组织中TUM2-PK mRNA的表达并研究其与临床病理特征间的联系。结果:TUM2-PK mRNA在肝癌组织和癌旁组织中的阳性表达率分别为83.33%(35/42)和57.14%(24/42),表达量为0.7230±0.6744和0.2594±0.2835,肝癌组织中TUM2-PK mRNA的阳性表达率及表达量显著高于癌旁组织(P<0.05)。TUM2-PK mRNA的表达与肝癌分化程度有关(P<0.05),而与肝癌的大小、门脉癌栓、淋巴转移、肝内转移、AFP水平及患者5年生存率无关(P>0.05)。结论:TUM2-PK mRNA的高表达提示肝癌的分化差。可作为评估肝癌恶性程度的生物学指标。     

Inhibition of tumour growth by lipoxygenase inhibitors.

The potential involvement of lipoxygenase metabolites in the tumour growth stimulatory activity of arachidonic and linoleic acid has been studied using the 5-lipoxygenase inhibitors, BWA4C, BWB70C and Zileuton. In vitro the former two agents were relatively potent inhibitors of growth of murine adenocarcinomas (MACs) with IC50 values < 10 microM, whereas Zileuton was less effective. In vivo studies showed BWA4C to be an effective inhibitor of the growth of both the MAC26 and MAC16 tumours at dose levels between 5 and 25 mg kg-1 (b.d.). The growth rate of the MAC26 tumour was also decreased by BWB70C at 25 mg kg-1, whereas lower doses were either ineffective or stimulated tumour growth. This differential effect of the 5-lipoxygenases inhibitors on tumour growth may arise from effects on the 12- and 15-lipoxygenase pathways. To quantify the effect cells were labelled with [3H]arachidonic acid and the biosynthesis of 5-, 12- and 15-hydroxyeicosatetraenoic acid (HETE) was analysed by high-performance liquid chromatography. All three agents caused a decrease in 5-HETE production, although the effect was less pronounced with Zileuton. In MAC26 cells both BWA4C and BWB70C caused a decrease in 12-HETE formation whereas Zileuton had no effect on the other lipoxygenase pathways. The inhibitory effect of these agents on cell growth may result from an imbalance of metabolism of arachidonic acid between the 5-, 12- and 15-lipoxygenase pathways.     

Effects of novel 5-lipoxygenase inhibitors on the incidence of pulmonary adenomas in the A/J murine model when administered via nose-only inhalation

The objective of this study was to determine the effects of 5-lipoxygenase (5-LO) inhibitors on the incidence of benzo(a)pyrene-induced pulmonary adenomas in female A/J mice. Two novel compounds, S-29606 and S-30621, and the Food and Drug Administration-approved Zileuton were investigated. S-29606 and S-30621 were selected from a group of similar active structures on the basis of local versus systemic 5-LO inhibitory activity. Preliminary studies found them to lack oral bioavailability, in direct contrast to Zileuton. Treatment was initiated 1 week following exposure to the carcinogen benzo(a)pyrene. Both S-29606 and S-30621 were dosed via nose-only inhalation 5 days a week, for 16 weeks, whereas Zileuton was administered orally. Dose levels for S-29606 and S-30621 were determined to be 220 and 430 microg/kg for the low- and high-dose groups, respectively, whereas the dose of Zileuton was 245 mg/kg. Both test compounds exhibited a significant reduction of pulmonary adenomas, compared with a positive control for high and low doses, P < 0.05. Additionally, a dose response for both S-29606 and S-30621 was observed when compared with placebo. Despite a dose 575 times greater than that of the novel test compounds, orally administered Zileuton did not produce a reduction in adenoma occurrence. The findings of this study offer compelling preliminary data for the use of S-29606 and S-30621 in further investigations of the treatment of pulmonary adenomas and support the use of inhalation drug delivery as an alternate to oral delivery for these compounds.     

Expression of L-type amino acid transporter 1 (LAT1) and 4F2 heavy chain (4F2hc) in liver tumor lesions of rat models.

BACKGROUND AND OBJECTIVES: It has been said that amino acid transporters play an important role in supplying nutrition to cells and for cell proliferation. In this study, we examined whether LAT1 and 4F2hc are closely related to tumor growth. METHODS: Rat colon cancer cells (RCN-9) were injected into the spleen of 12 male rats (inbred F344/DuCrj). In each rat, liver samples including tumor lesions were immunostained with anti-LAT1 and anti-4F2hc antibodies. The staining area of LAT1 and 4F2hc tumor lesions was calculated by computer analysis. RESULTS: Sixty-eight tumor nodules were observed in 12 livers. Out of the 68 tumor nodules, 36 nodules (52.9%) indicated a positive staining of LAT1 and 32 (47.1%) had a negative staining of LAT1. However, the LAT1 expression was scarcely detected in non-tumor areas. In terms of the 4F2hc expression, there were 56 nodules (82.4%) with 4F2hc positive and 12 (17.6) with 4F2hc negative. In addition, the expression of 4F2hc in non-tumor areas was almost the same as the expression of 4F2hc in tumor lesions. The average tumor size of the group with LAT1 positive and 4F2hc positive (n = 31) was 0.845 +/- 0.232 mm(2), which was significantly larger than that of the group with LAT1 negative and 4F2hc negative group (n = 7) (0.090 +/- 0.028 mm(2)) or the group with LAT1 positive and 4F2hc negative (n = 5) (0.097 +/- 0.025 mm(2)), respectively (P = 0.0017, P = 0.007). CONCLUSION: LAT1 was related to tumor growth. We think that LAT1 can possibly enhance its ability to promote tumor growth in cooperation with 4F2hc.     

Impact of thymidine phosphorylase surexpression on fluoropyrimidine activity and on tumour angiogenesis

Tumoral thymidine phosphorylase (TP) appears to play a dual role by being involved in neoangiogenesis and by activating 5FU prodrugs at the tumoral target site. The aim of the study was to investigate more thoroughly these potential physiological and pharmacological roles of TP. A rat carcinoma cell line (PROb) was transfected with TP/PD-ECGF in order to study the effect of the overexpression of this enzyme (1) on the sensitivity of cells to 5'DFUR and 5FU in vitro and (2) on tumour growth in vivo by using a syngenic tumour model in the BDIX rat (hepatic tumours, sub-cutaneous tumours). Cytotoxic effects of 5'DFUR, and to a lesser extent those of 5FU, were enhanced in TP clones as compared to control cells: there was a highly significant correlation between TP activity and in vitro sensitivity to 5'DFUR (r2= 0.91, P = 0.0002, n = 8) and, to a lesser extent, to 5FU (r2= 0.49, P = 0.053, n = 8). The impact of TP transfection on tumour growth was relatively modest and concerned only the initial stages of tumour expansion. Staining of TP tumours for endothelial (factor VIII) cells was always higher than controls. The staining ratio (TP/controls) tended to be reduced as tumours increased in size. The stability of TP expression was checked both in vitro (TP activity measurement) and in vivo (RT-PCR determinations) and there was no loss of TP expression over time which could be advanced to explain the progressive weakening of the impact of TP overexpression on both tumour growth and neoangiogenesis.     

Overexpression of 5-lipoxygenase and cyclooxygenase 2 in hamster and human oral cancer and chemopreventive effects of zileuton and celecoxib.

PURPOSE: Previous studies have suggested an important role of aberrant arachidonic acid metabolism, especially the cyclooxygenase (Cox) pathway, in oral carcinogenesis. However, it is unknown whether the 5-lipoxygenase (5-Lox) pathway contributes to oral carcinogenesis, and whether combination of inhibitors of both pathways may have synergistic or additive effects of chemoprevention. EXPERIMENTAL DESIGN: 5-Lox expression was examined in 7,12-dimethylbenz[a]anthracene (DMBA)-induced hamster and human oral cancer tissues by immunohistochemistry, and Cox2 expression was investigated in hamster oral tissues using in situ hybridization. Zileuton (a specific 5-Lox inhibitor) and celecoxib (a specific Cox2 inhibitor), either alone or in combination, were investigated for their chemopreventive effects on the DMBA-induced hamster model at the post-initiation stage through topical application. RESULTS: 5-Lox was overexpressed during oral carcinogenesis in hamsters and humans, as well as Cox2 in the hamster tissues. In a chemoprevention study using the post-initiation DMBA model, incidence of hamster oral squamous cell carcinoma was reduced from 76.9% (20 of 26) to 45.8% (11 of 24, P < 0.05) and 32.1% (9 of 28, P < 0.01) by 3% and 6% topical zileuton, respectively; and to 57.6% (15 of 26, P > 0.05) and 50% (12 of 24, P < 0.05) by 3% and 6% topical celecoxib, respectively. When used in combination, celecoxib and zileuton (3% of each) had an additive inhibitory effect on the incidence of squamous cell carcinoma (36%, 9 of 25, P < 0.01). Other pathologic variables and the levels of leukotriene B4 and prostaglandin E2 of the hamster tissues were reduced as well. CONCLUSIONS: The results clearly showed that both 5-Lox and Cox2 played important roles in oral carcinogenesis. Zileuton and celecoxib prevented oral carcinogenesis at the post-initiation stage through their inhibitory effects on arachidonic acid metabolism.     

雌激素对垂体催乳素细胞致瘤机制的探讨

【摘要】 背景与目的： 探讨雌激素诱发大鼠垂体催乳素(PRL)瘤的发病机制。 材料与方法： 制备雌激素诱发大鼠PRL瘤模型: 成年雄性Wistar大鼠16只,随机分为对照组(n=8)和PRL瘤组(n=8) 。对照组皮下植入空白硅胶管,PRL瘤组皮下植入含有乙烯雌酚的硅胶管,用药8周后乙醚麻醉大鼠,心脏穿刺取血,分离血清,并开颅摘取垂体。采用放免法测定大鼠血清PRL水平;垂体称重并做组织病理学观察;用免疫组织化学方法检测垂体组织PRL蛋白的表达和分布;用RT_PCR方法检测c_fos 和垂体肿瘤转化基因(PTTG)在PRL瘤组织中的表达,计算机凝胶成像系统分析其表达量。 结果： 雌激素作用8周后,根据大鼠垂体重量以及组织学和免疫组化的改变,证实已诱发出PRL瘤模型。在PRL瘤组中,c_fos和PTTG的表达量均明显高于对照组,二者差异具有统计学意义彩 (P<0.001)。 结论： 雌激素刺激垂体PRL细胞表达癌基因c_fos以及PTTG,在雌激素诱发大鼠PRL细胞增生以至最终形成PRL瘤的过程中起一定的作用。     

Osteopontin and interleukin-8 expression is independently associated with prostate cancer recurrence.

PURPOSE: Lack of reliable biomarkers limits accurate prediction of prostate-specific antigen biochemical recurrence (disease progression) in prostate cancer. The two inflammatory chemokines, osteopontin and interleukin-8 (IL-8), are associated with tumor angiogenesis and metastasis. We investigated whether osteopontin and IL-8 expression in prostate cancer correlates with disease progression. EXPERIMENTAL DESIGN: Archival prostatectomy specimens (n = 103) were obtained from patients with minimum 72-month follow-up. Osteopontin and IL-8 expression was evaluated by immunohistochemistry and graded for intensity and the area. Association of osteopontin and IL-8 staining with biochemical recurrence was evaluated by univariate and multivariate models. RESULTS: In tumor cells, osteopontin and IL-8 staining was higher in the recurred group (203.2 +/- 78.4; 181.1 +/- 89.3) than in the nonrecurred group (122.7 +/- 76.6; 96.4 +/- 85.6; P < 0.001). Higher osteopontin and IL-8 staining was also observed in benign areas adjacent to tumor in the recurred group, than in nonrecurred group. In univariate analysis, except age, all preoperative and postoperative variables and osteopontin and IL-8 staining scores were significantly associated with biochemical recurrence (P < 0.05). In multivariate analysis, margin status and osteopontin staining independently associated with biochemical recurrence within 72 months. Osteopontin, either alone or with IL-8 and seminal vesicle invasion, was a significant variable in predicting biochemical recurrence within 24 months. Osteopontin and IL-8 staining predicted recurrence with high sensitivity (75.5%; 73.6%) and specificity (76%; 70.6%). CONCLUSION: In prostatectomy specimens, osteopontin expression is independently associated with biochemical recurrence. Both osteopontin and IL-8 may be predictors of early disease progression.