血清IGF-1和IGFBP-3检测在前列腺癌和前列腺增生诊断中的临床意义

目的 :探讨胰岛素样生长因子 1(IGF 1)、胰岛素样生长因子结合蛋白 3(IGFBP 3)对前列腺癌 (PCa)早期诊断的临床意义。方法 :采用免疫放射分析法 (IRMA)检测 32例PCa、33例良性前列腺增生 (BPH)患者和 18例健康人血清IGF 1、IGFBP 3水平。结果 :PCa组血清IGF 1为 (4 89.9± 171.9) μg/L ,明显高于BPH组 (2 81.2±70 .4 ) μg/L和健康组 (2 5 9.6± 6 4 .8) μg/L ,差异有统计学意义 (P <0 .0 1) ,BPH组与健康组比较差异无统计学意义(P >0 .0 5 )。PCa组血清IGFBP 3为 (6 8.1± 12 .6 ) μg/L ,明显高于BPH组 (5 1.1± 8.8) μg/L和健康组 (4 8.9± 8.1)μg/L ,差异有统计学意义 (P <0 .0 1) ,BPH组与健康组比较差异无统计学意义 (P >0 .0 5 )。结论 :IGF 1、IGFBP 3有可能成为临床上检测PCa的新的瘤标。     

基质交联分子1对前列腺癌PC-3细胞凋亡相关蛋白表达的影响

目的:观察抑制基质交联分子1(STIM1)对前列腺癌PC-3细胞凋亡相关蛋白表达的影响。方法:将携带STIM1基因的小干扰RNA(shRNA)慢病毒载体STIM1-pGCSIL-GFP转染人激素非依赖性前列腺癌PC-3细胞,3d后荧光倒置显微镜观察转染效率;1周后RT-PCR及Western印迹验证STIM1抑制表达有效性,并采用Western印迹检测PC-3细胞中凋亡相关蛋白Bcl-2、Bax,survivin、激活型Caspase-3的表达水平。结果:倒置显微镜观察发现PC-3细胞病毒转染效率80%。转染1周后,RT-PCR及Western印迹显示STIM1被有效抑制。抑制STIM1表达后,对照组Bcl-2/Bax比率为1.24,干扰组PC-3细胞Bcl-2/Bax比率为0.31,比率显著下降;干扰组PC-3细胞survivin表达明显降低,相对表达量为对照组的0.14倍;Caspase-3裂解激活相对表达量为对照组的1.52倍(P0.05)。结论:STIM1在前列腺癌PC-3细胞中可视为致癌基因,抑制其表达可通过下调Bcl-2/Bax比率,降低survivin表达,激活Caspase-3级联通路,诱导细胞凋亡。     

IGF-Ⅰ、IGF-Ⅱ及IGF-ⅠR在前列腺癌中的表达和意义

目的 研究Ⅰ型胰岛素样生长因子(IGF-Ⅰ)、Ⅱ型胰岛素样生长因子(IGF-Ⅱ)和胰岛素样生长因子Ⅰ受体(IGF-IR)在中国患者前列腺癌组织中的表达情况及其与前列腺癌生物学行为的关系.方法 应用免疫组织化学法检测IGF-Ⅰ、IGF-Ⅱ和IGF-1R在良性前列腺增生及前列腺癌组织中的表达情况,实验所得数据用SPSS 12.0统计软件处理.计数资料采用直接概率法和四格表x2检验,当n＞40 fH有1≤T＜5时,用校正的卡方检验,P＜0.05为差异有显著性的标准.Spearman等级相关方法分析免疫组化结果与临床病理因素的关系.结果 IGF-Ⅰ在良性前列腺增生和前列腺癌组织中的阳性表达率分别为26.67%(8/30)、65.12%(28/43)(P＜0.05);IGF-Ⅱ在良性前列腺增生和前列腺癌组织中的表达率分别为36.67%(11/30)、72.09%(31/43)(P＜0.05);IGF-IR在良性前列腺增生和前列腺癌组织中的表达率分别为:43.33%(13/30)、79.07%(34/43)(P＜0.05).结论 IGF-Ⅰ、IGF-Ⅱ和IGF-ⅠR与前列腺癌的发生相关,IGF-Ⅰ、IGF-Ⅱ与前列腺癌的发展呈正相关,IGF-ⅠR与前列腺癌的发展不相关.     

正常前列腺不同区带及癌组织来源原代基质细胞的生物学特性及其对前列腺癌细胞株C4-2B的作用

目的 比较正常前列腺外周带来源原代基质细胞(NPPF),移行带来源原代基质细胞(NPTF)及前列腺癌组织来源原代基质细胞(CAF)的生物学特性差异,以及它们对前列腺癌细胞株C4-2B的不同影响.方法 苏木素-伊红(HE)染色鉴定前列腺不同区带及癌组织的组织学特征差异;原代培养NPPF、NPTF、CAF,免疫细胞化学染色观察其波形蛋白(Vimentin)、平滑肌动蛋白(SMA)及前列腺特异性抗原(PSA)的表达,生长曲线比较其增殖能力,流式细胞仪检测比较其凋亡率,透射电镜观察比较其超微结构;建立不同来源原代基质细胞与C4-2B细胞株共培养系统,比较不同来源基质细胞对C4-2B细胞增殖(MTT)和凋亡(FCM)方面的影响.结果 NPPF、NPTF、CAF的生长、凋亡、超微结构及对C4-2B细胞的生物学影响存在明显差异,其生长速度依次递增,凋亡率分别为(9.25±2.24)%、(5.98±0.74)%、(2.63±0.96)%,透射电镜提示CAF蛋白质合成最旺盛;C4-2B细胞与基质细胞体外共培养后出现增殖加速,凋亡减少,MTT结果显示共培养2 d后吸光度分别0.540、0.471、0.632,共培养4 d后吸光度分别为0.554、0.488、0.670,P均<0.05;FCM结果显示与对照组(10.32±0.43)%比较,CAF对C4-2B细胞凋亡的抑制能力最强(3.36±0.17)%,其次是NPPF(5.97±0.70)%和NPTF(8.01±0.22)%,P<0.05.结论 NPPF、NPTF、CAF的生物学特性存在明显差异,这种差异可能是导致前列腺增生或前列腺癌发生发展的重要原因.     

心肌梗死后残存心肌细胞肥大性改变及其IGF-1、IGF-1R的表达

目的观察心肌梗死后残存心肌细胞肥大性改变及其胰岛素样生长因子-1(IGF-1)、胰岛素样生长因子-1受体(IGF-1R)的表达,探讨梗死心肌心肌细胞肥大的机制。方法取急性心肌梗死后2周、4周、8周的梗死心肌,制备单心肌细胞悬液,采用激光共聚焦扫描显微镜(Confocal)检测心肌细胞体积,采用免疫组织化学法检测心肌细胞IGF-1、IGF-1R的表达。结果急性心肌梗死8周梗死区域心肌细胞体积较正常区域心肌细胞增大(37563.93±6176.79μm3vs.28638.61±6890.89μm3,t=4.840,P=0.020),细胞肥大以宽度和厚度为主;梗死区域心肌细胞可见IGF-1染色阳性颗粒,IGF-1表达高于正常区域心肌细胞(79.58±4.57vs.64.12±3.91,t=27.564,P=0.002);IGF-1R主要分布于心肌细胞膜,梗死区域心肌细胞IGF-1R表达高于正常区域心肌细胞(67.02±2.56vs.66.73±3.49,t=3.845,P=0.042),其中以急性心肌梗死4周IGF-1R表达最高。结论心肌梗死后残存心肌细胞分泌IGF-1和IGF-1R,并可能在促使残存心肌细胞肥大中起了积极作用。     

胰岛素样生长因子系统与前列腺癌

胰岛素样生长因子（IGF）是一组重要的肽类生长因子,在前列腺癌的发生发展过程中发挥着重要的作用。目前对IGF系统与前列腺癌的关系研究取得较大进展,与前列腺癌的发病机制密切相关,为前列腺癌的诊断及治疗提供了重要的参考作用。但IGF系统十分复杂,其对前列腺癌的确切作用目前仍有争论,需要做进一步的实验和临床研究。     

胰岛素样生长因子-1及其受体在前列腺癌组织的表达及其临床意义

目的:探讨胰岛素样生长因子-1(IGF-1)及其受体(IGF-1R)在前列腺癌(PCa)组织中的表达及其临床意义。方法:取2017年1月至2019年12月河北医科大学第三医院泌尿外科的41例前列腺癌患者的前列腺组织作为实验组;并收集21例良性前列腺增生(BPH)患者前列腺组织当做对照组;并通过前列腺特异性抗原(PSA)...     

骨髓基质细胞向成骨细胞诱导分化的研究进展

骨髓基质细胞(bone marrow stromal cells,BMSC)是位于骨髓中的一种干细胞,因在体外经适当的培养条件可以向成骨细胞、成软骨细胞、脂肪细胞、成肌细胞等多种间充质来源的组织细胞分化,又被称为间充质干细胞(mesenchymal stem cell).骨髓基质细胞位于骨髓,具有取材方便,易于体外扩增,自体移植无免疫排斥性等优点,是骨组织工程中种子细胞的理想来源.骨髓基质细胞,其向成骨细胞的定向诱导分化将是关键的一步.笔者就骨髓基质细胞向成骨细胞诱导分化有关影响因素的研究进展作一综述.     

胰岛素样生长因子-1抑制人椎间盘细胞凋亡及促进基质代谢的研究

胰岛素样生长因子1(IGF1)在软骨形成和软骨自稳态调节中起关键作用。有研究表明,IGF1能刺激软骨细胞分裂增殖,并可延缓及阻止多种体细胞的凋亡[1]。我们通过体外培养人椎间盘细胞,观察不同浓度胰岛素生长因子对椎间盘细胞凋亡、增殖及细胞外基质代谢的影响。一、材料与方法1.材料:取自8例施行脊柱侧凸前路松解术的患者,其中男4例,女4例,年龄12～17岁,平均15.2岁。2.椎间盘细胞原代培养及形态学观察:用倒置相差显微镜观察原代及传代培养的椎间盘细胞。3.IGF1对人椎间盘细胞凋亡及细胞外基质代谢的影响:Tunel法检测椎间盘细胞凋亡情况;收…     

性激素受体及相关生长因子在前列腺基质细胞中的表达

我们采用半定量ＲＴ ＰＣＲ方法观察了性激素受体及相关生长因子在ＢＰＨ体外细胞模型前列腺基质细胞中的表达 ,以及在前列腺基质细胞分化和逆分化中的表达变化。报告如下。材料与方法　前列腺切除术或膀胱全切除术组织标本 5例。标本切成 1～3ｍｍ3 小块 ,洗涤。在组织消化液中 ,37℃水浴轻搅 4～ 5ｈ。消化后的细胞悬液通过 10 0 μｍ滤网 ,洗涤后 ,80 0×ｇ离心10ｍｉｎ收集细胞沉淀物。细胞沉淀物用平滑肌细胞 (ＳＭＣ)培养液悬浮 ,并通过40 μｍ微孔过滤 ,台盼蓝染色细胞。每 ｇ前列腺组织大约获得 3× 10 6个活细胞。消化所得的细胞悬…     

The differential effects of prostate stromal cells derived from different zones on prostate cancer epithelial cells under the action of sex hormones

It is well known that prostate cancer (PCa) occurs predominantly in the peripheral zone (PZ), whereas benign prostatic hyperplasia (BPH) typically develops in the transition zone. To identify possible mechanisms underlying zonal differences, we compared the effects of prostate stromal cells derived from the peripheral zone (PZsc) and the transition zone (TZsc) on a PCa epithelial cell line (PC3) in the presence of sex hormones. First, we observed that androgen receptor (AR) mRNA was more highly expressed in PZsc than TZsc when the cells were treated with dihydrotestosterone (DHT) and β-oestradiol (E2) (P<0.05). By ELISA, we looked for differences in the secretion of peptide growth factors from PZsc and TZsc. We found that keratinocyte growth factor (KGF) secretion increased with increasing concentrations of DHT (P<0.01) and was higher in PZsc than TZsc. Under treatment with DHT plus E2, PZsc secreted more transforming growth factor-β1 (TGF-β1) than TZsc, but this pattern was reversed when the cells were treated with E2 only. With increasing concentrations of DHT, insulin-like growth factor-1 (IGF-1) secretion increased in PZsc but decreased in TZsc. To further characterize the effects of PZsc and TZsc on PC3 cells, we developed a coculture model and performed MTT assays, Western blot analysis and real-time RT-PCR. We found that PZsc promoted PC3 cell proliferation and progression better than TZsc, particularly when treated with 10 nmol l⁻¹ DHT plus 10 nmol l⁻¹ E2. In conclusion, our data suggest that PZsc may have a greater capacity to induce PCa development and progression than TZsc via growth factors regulated by sex hormones. These findings provide possible mechanisms underlying zonal differences in prostate diseases, which may aid the search for novel therapeutic targets for PCa.     

Serum measurements of testosterone, insulin-like growth factor 1, and insulin-like growth factor binding protein-3 in the diagnosis of prostate cancer among Korean men

Aim： To investigate the relationships of serum testosterone, insulin-like growth factor （IGF）- 1 and IGF-binding protein （IGFBP）-3 levels with prostate cancer risk and also with known prognostic parameters of prostate cancer in Korean men who received radical retropubic prostatectomy （RRP） for clinically-localized prostate cancer. Methods： Serum levels of total testosterone, free testosterone, IGF-1 and IGFBP-3 were determined in 592 patients who subsequently received prostate biopsy. Results were compared between patients who eventually received RRP for prostate cancer （n = 159） and those who were not diagnosed with prostate cancer from biopsy （control group, n = 433）. Among the prostate cancer only patients, serum hormonal levels obtained were analyzed in relation to serum prostate specific antigen （PSA）, pathological T stage and pathological Gleason score. Results： Prostate cancer patients and the control group demon- strated no significant differences regarding serum levels of total testosterone, free testosterone, IGF-I and IGFBP-3 across the different age groups. Among the cancer only patients, no significant associations were observed for serum levels of total testosterone, free testosterone, IGF-1 and IGFBP-3 levels with pathological T stage, pathological Oleason score and preoperative PSA. Conclusion： Our data indicate that simple quantifications of serum testosterone and IGF-1 along with IGFBP-3 levels might not provide useful clinical information in the diagnosis of clinically localized prostate cancer in Korean men. Also, our results suggest that serum levels of testosterone, IGF-1 and IGFBP-3 might not be significantly associated with known prognostic factors of clinically localized prostate cancer in Korean men. （Asian J Androl 2008 Mar; 10： 207-213）     

Differences in phenotype and gene expression of prostate stromal cells from patients of varying ages and their influence on tumour formation by prostate epithelial cells

Prostate cancer (PCa) is an age-related disease, and the stromal microenvironment plays an important role in prostatic malignant progression. However, the differences in prostate stromal cells present in young and old tissue are still obscure. We established primary cultured stromal cells from normal prostatic peripheral zone (PZ) of donors of varying ages and found that cultured stromal cells from old donors (PZ-old) were more enlarged and polygonal than those from young donors (PZ-young). Furthermore, based on immunocytochemical and ultrastructural analysis, the components of stromal cells changed from a majority of fibroblasts to a mixture of fibroblasts and myofibroblasts with increasing donor age. Using a three-dimensional in vitro culture system, we found that PZ-old stromal cells could enhance the proliferation, migration and invasion of cocultured benign BPH-1 and PC-3 cells. Using an in vivo tissue recombination system, we also found that PZ-old stromal cells are more effective than PZ-young cells in promoting tumour formation by BPH-1 cells of high passage (>100) and PC-3 cells. To probe the possible mechanism of these effects, we performed cDNA microarray analysis and profiled 509 upregulated genes and 188 downregulated genes in PZ-old cells. Among the changed genes, we found genes coding for a subset of paracrine factors that are capable of influencing adjacent epithelial cells; these include hepatocyte growth factor (HGF), fibroblast growth factor 5 (FGF5), insulin-like growth factor 2 (IGF2), insulin-like growth factor-binding protein 4 (IGFBP4), IGFBP5 and matrix metallopeptidase 1 (MMP1). Changes in the expression of these genes were further confirmed by quantitative real-time polymerase chain reaction (PCR), Western blotting and enzyme-linked immunosorbent assays. Overall, our findings indicate that stromal cells from prostate PZ of old donors are more active than similar cells from young donors in promoting the malignant process of adjacent epithelial cells. This finding hints at a new potential strategy for the prevention of PCa.     

Investigations of prostate epithelial stem cells and prostate cancer stem cells

Although both prostate epithelial stem cells and prostate cancer stem cells are implicated in the differentiation of the normal prostate gland and carcinogenesis of prostate cancer, there has, until recently, been little information regarding their biology. This review summarizes the recent advancements in cell biological research including various in vitro culture systems that have offered the characterization and isolation of prostate epithelial stem cells and prostate cancer stem cells. In addition, the stromal niche or microenvironment of stem cells plays an essential role in proliferation and differentiation of normal stem cells. Stroma surrounding cancer cells, which also provide another unique niche, may involve the initiation and development of cancer stem cells. Investigation of stem cells and their microenvironments in the prostate should lead to the elucidation of biological features and the development of novel treatments for prostate cancer.     

Tumor formation of prostate cancer cells influenced by stromal cells from the transitional or peripheral zones of the normal prostate

This study was designed to investigate the different involvements of prostatic stromal cells from the normal transitional zone (TZ) or peripheral zone (PZ) in the carcinogenesis of prostate cancer (PCa) epithelial cells (PC-3) in vitro and in vivo co-culture models. Ultra-structures and gene expression profiles of primary cultures of human prostatic stromal cells from the normal TZ or PZ were analyzed by electron microscopy and microarray analysis. In vitro and in vivo co-culture models composed of normal TZ or PZ stromal cells and human PCa PC-3 cells were established. We assessed tumor growth and weight in the in vivo nude mice model. There are morphological and ultra-structural differences in stromal cells from TZ and PZ of the normal prostate. In all, 514 differentially expressed genes were selected by microarray analysis; 483 genes were more highly expressed in stromal cells from TZ and 31 were more highly expressed in those from PZ. Co-culture with PZ stromal cells and transforming growth factor-β1 (TGF-β1) increased the tumor growth of PC-3 cells in vitro and in vivo, as well as Bcl-2 expression. On the other hand, stromal cells of TZ suppressed PC-3 cell tumor growth in the mouse model. We conclude that ultra-structures and gene expression differ between the stromal cells from TZ or PZ of the normal prostate, and stroma–epithelium interactions from TZ or PZ might be responsible for the distinct zonal localization of prostate tumor formation.     

Stromal cells of the human prostate: Initial isolation and characterization

The present study was conducted to isolate and to characterize stromal cells from the human prostate and to study the effects of androgen and different growth factors in this model system. Benign prostatic hyperplasia (BPH) tissue samples were obtained from transurethral resection of the prostate (TURP). Tissue specimens were mechanically and enzymatically dissociated by treatment with DNAse and collagenase. Epithelial cells were separated from stromal cells by discontinuous Percoll gradient centrifugation. The stromal cells obtained were cultured in phenol red-free RPMI-1640 supplemented with 10% fetal bovine serum. Immunocytochemical analysis revealed that the stromal cell cultures were composed of both smooth muscle cells and fibroblasts. The short and broad, smooth muscle cells were identified by using an antibody directed against α-smooth muscle actin. The thin and elongated fibroblasts stained positively for prolyl 4-hydroxylase. Smooth muscle cells were the predominant cell type in the present investigation. Typical cultures contained up to 99% of cells staining positively for α-smooth muscle actin. The prostate smooth muscle cultures were treated with dihydrotestosterone (DHT), bovine pituitary extract (BPE), basic fibroblast growth factor (bFGF) and transforming growth factor-β (TGF-β). When cells were cultured in serum free RPMI-1640 supplemented with ITS+ (insulin, transferrin, and selenious acid) no significant (P > 0.05) stimulative effect after treatment with DHT or TGF-β was detectable. Basic FGF had a slight but significant (P < 0.05) mitogenic effect in medium supplemented with ITS+. In the presence of 10% charcoal-stripped fetal bovine serum (cFBS) DHT, at a concentration of 0.1 nM, was able to cause a slight but significant (P < 0.05) mitogenic effect on BPH smooth muscle cells growth. Basic FGF was able to stimulate BPH smooth muscle cells in a concentration-dependent fashion. The combination of DHT and 0.1 ng/ml bFGF was able to increase the proliferation of prostate smooth muscle cells above either agents alone. Addition of BPE to serum free RPMI-1640 caused a significant (P < 0.05) stimulation of cell proliferation in a concentration-dependent fashion. Addition of TGF-β to serum or BPE containing RPMI-1640 caused a significant (P < 0.05) inhibition of cell proliferation in a concentration-dependent fashion. TGF-β was cytostatic to the benign prostatic smooth muscle cells only in the presence of media containing growth stimulating factors found in charcoal-stripped serum or in bovine pituitary extract. These results demonstrated that stromal fraction isolated from BPH specimens was composed of both fibroblasts and smooth muscle cells. These cells could be cultured and were able to respond to various growth stimulatory and inhibitory agents. © 1996 Wiley-Liss, Inc.     

Insulin-like growth factor-1 and insulin-like growth factor binding protein-3 for prostate cancer detection in patients undergoing prostate biopsy

PURPOSE: Laboratory and epidemiological studies suggest that high circulating insulin-like growth factor (IGF)-1 and low IGF binding protein-3 are associated with increased prostate cancer risk. However, the usefulness of serum IGF-1 or IGF binding protein-3 for predicting pathology results in men undergoing prostate biopsy is unclear. We examined the relationships of serum IGF-1, IGF binding protein-3 and the results of prostate biopsy. MATERIALS AND METHODS: A total of 652 consecutive patients with elevated serum prostate specific antigen (PSA) or abnormal digital rectal examination who were referred for transrectal ultrasound sextant prostate needle biopsy underwent blood sampling before biopsy. PSA, free PSA, IGF-1 and IGF binding protein-3 were measured. There were 244 men (37.4%) with cancer and 408 controls with benign conditions. RESULTS: Mean IGF-1 plus or minus SD in the cancer and control groups was 176.1 +/- 58.3 and 178.7 +/- 54.7 ng./ml., respectively (p = 0.57). Mean IGF binding protein-3 in the cancer and control groups was 2,724 +/- 647 and 2,673 +/- 589 ng./ml., respectively (p = 0.3). Adjustment for age and PSA showed significantly lower IGF-1 in cancer cases, while IGF binding protein-3 was not significant. ROC values were significantly higher for free-to-total PSA and PSA than for crude and age adjusted IGF-1 and IGF binding protein-3. CONCLUSIONS: Our data indicate that serum IGF-1 or IGF binding protein-3 does not predict the results of prostate biopsy in men with elevated PSA or abnormal digital rectal examination. This finding implies that while there is evidence that the IGF-1 level is a risk factor for prostate cancer, neither IGF-1 nor IGF binding protein-3 can be used as a tumor marker for this disease.