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ETAR 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。     

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。     

姜黄素对雄激素非依赖性前列腺癌细胞PC-3及血管内皮生长因子的影响

目的：研究姜黄素对雄激素非依赖性前列腺癌细胞株PC-3细胞体外作用及其对血管内皮生长因子（VEGF）表达的影响，探讨其抗肿瘤的作用机制。方法：分别用0、6．25、12．5、25、50μmol／L浓度的姜黄素作用于PC-3细胞，12、24、36、48、72、96h后台盼蓝拒染法、四甲基偶氮唑蓝（MTT）法检测细胞生长活性；24h后流式细胞仪测定细胞周期及凋亡的变化，透射电镜观察细胞超微结构变化；半定量RT-PCR法检测PC-3细胞内VEGF mRNA的表达；ELISA检测细胞上清液中VEGF浓度。结果：姜黄素能显著抑制PC-3细胞的增殖，呈剂量与时间依赖性，不同浓度姜黄素组之间及不同时间组之间差异均有统计学意义（P〈0．01）。不同浓度姜黄素诱导PC-3细胞出现剂量依赖性G2／M期阻滞（P〈0．01），且各浓度组凋亡细胞比例均显著高于空白对照组（P〈0．01），差异有统计学意义；姜黄素作用24h后PC-3细胞出现凋亡的形态学改变；PC-3细胞内VEGF mRNA的表达和细胞上清液中VEGF呈剂量依赖性降低。结论：姜黄素能显著抑制体外PC-3细胞的生长，并促进其G2／M期阻滞和凋亡，VEGF mRNA及蛋白的表达也明显降低，可能是其抑制肿瘤和血管生长的机制之一。     

姜黄素对雄激素非依赖性前列腺癌细胞PC-3及血管内皮生长因子的影响

目的:研究姜黄素对雄激素非依赖性前列腺癌细胞株PC-3细胞体外作用及其对血管内皮生长因子(VEGF)表达的影响,探讨其抗肿瘤的作用机制。方法:分别用0、6.25、12.5、25、50μmol/L浓度的姜黄素作用于PC-3细胞,12、24、36、48、72、96h后台盼蓝拒染法、四甲基偶氮唑蓝(MTT)法检测细胞生长活性;24h后流式细胞仪测定细胞周期及凋亡的变化,透射电镜观察细胞超微结构变化;半定量RT-PCR法检测PC-3细胞内VEGFmRNA的表达;ELISA检测细胞上清液中VEGF浓度。结果:姜黄素能显著抑制PC-3细胞的增殖,呈剂量与时间依赖性,不同浓度姜黄素组之间及不同时间组之间差异均有统计学意义(P<0.01)。不同浓度姜黄素诱导PC-3细胞出现剂量依赖性G2/M期阻滞(P<0.01),且各浓度组凋亡细胞比例均显著高于空白对照组(P<0.01),差异有统计学意义;姜黄素作用24h后PC-3细胞出现凋亡的形态学改变;PC-3细胞内VEGF mRNA的表达和细胞上清液中VEGF呈剂量依赖性降低。结论:姜黄素能显著抑制体外PC-3细胞的生长,并促进其G2/M期阻滞和凋亡,VEGFmRNA及蛋白的表达也明显降低,可能是其抑制肿瘤和血管生长的机制之一。     

Differential Expression of Hepatocyte Growth Factor in Papillary and Nodular Tumors of the Bladder

Background: The aim of this study was to investigate the expression of hepatocyte growth factor (HGF) and its receptor (c-met) during bladder tumorigenesis.
Methods: HGF and c-met expression were analyzed immunohistochemically in matched samples of normal, dysplastic, and carcinoma specimens from 49 human bladders resected at the time of radical cystectomy for nonmetastatic transitional cell carcinoma (TCC). The tumors were composed of papillary (n = 22), nodular (n = 16) or mixed papillary and nodular (mixed; n = 11) components. Results: The normal urothelium showed no significant immunoreactivity to HGF. Expression of HGF was observed in 45.5%, 77.3% and 90.9% of specimens demonstrating mild, moderate, and severe dysplastic lesions adjacent to papillary TCCs, respectively, whereas all of the papillary TCC samples were positive for HGF. No immunoreactivity for HGF was found in dysplastic lesions from nodular tumors, and only 2 specimens had positive immunostainingfor HGF in the tumor areas (1 showed weak immunostainingand 1 showed HGF immunostainingonly in the deeper invasive compartment). Additionally, 3 nodular lesions taken from mixed tumors showed weak immunostaining for HGF while the concurrent papillary lesions were HGF-positive. There was a significant difference of HGF immunoreactivity between papillary and nodular tumors (P < 0.01). c-met immunostaining was consistently detected in all specimens. HGF and c-met immunoreactivity did not significantly correlate with tumor stage and grade, nor with overall patient survival irrespective of the tumor growth pattern.
Conclusion: These results suggest that HGF expression may play a significant role in the development of papillary TCC.     

前列腺癌粒细胞-巨噬细胞集落刺激因子受体表达的研究

目的 :研究粒细胞 巨噬细胞集落刺激因子 (GM CSF)受体在前列腺癌组织和 4种肿瘤细胞株中的表达及其意义。　方法 :应用SP免疫组织化学方法研究 4 8例前列腺癌和 2 0例良性前列腺增生石蜡标本中GM CSF受体α亚单位表达 ;Western印迹方法检测慢性粒细胞性白血病细胞株K5 6 2 ,乳腺癌细胞株MCF 7,转移性前列腺癌细胞株PC 3M和急性早幼粒细胞性白血病细胞株HL 6 0GM CSF受体α亚单位的表达。　结果 :4 8例前列腺癌中GM CSF受体α亚单位表达阳性病例 38例 ,阳性率为 79.2 %。其中Gleason 2～ 4分者 15例 ,5～ 8分 14例 ,9～ 10分 9例 ,GM CSF受体的阳性率与前列腺癌Gleason评分相关 ;2 0例良性前列腺增生中GM CSF受体α亚单位表达 6例 ,阳性率为 30 .0 %。 4种肿瘤细胞株K5 6 2、HL 6 0、PC 3M和MCF 7均表达GM CSF受体α亚单位。　结论 :GM CSF受体表达是前列腺癌的重要特征 ,GM CSF可能参与前列腺癌的增殖过程。     

端粒酶逆转录酶与血管内皮细胞生长因子在前列腺癌中的表达及其相关性的研究

目的:探讨端粒酶逆转录酶(TRT)、血管内皮细胞生长因子(VEGF)在前列腺癌(PCa)中的表达及两者的相关性。方法:应用免疫组化染色(SP)法结合计算机辅助图像分析方法,对30例病理证实为PCa和30例良性前列腺增生(BPH)穿刺标本中的TRT、VEGF的表达进行半定量分析。结果:30例PCa标本中,TRT、VEGF阳性表达率分别是63.3%(19/30)和76.7%(23/30);30例BPH标本中,TRT、VEGF阳性表达率分别是16.7%(5/30)和46.7%(14/30),组间差异均有显著性意义(P<0.05)。TRT与VEGF的表达显著相关(r=0.833 3,P<0.05)。结论:TRT、VEGF的表达可能是PCa的恶性表型,两者的表达具有显著相关性,但其确切机制还需深入研究。     

Expression of Hepatocyte Growth Factor in Chronic Hepatitis: Effect of Hemodialysis

Background and aims. Elevated serum levels of hepatocyte growth factor (HGF) have been documented in patients on chronic hemodialysis (HD) and with chronic hepatitis (CH). The aim of this study was to determine the presence and density of HGF in liver tissue and the source of this factor in HD patient with CH.?Material and methods. Seventy-five hepatic needle-biopsy specimens were investigated, 11 from CH patients (Group 1) and 44 from CH patients on HD (Group 2), plus 20 liver biopsies obtained from healthy individuals as controls (Group 3). All specimens were analyzed immunohistochemically with an HGF antibody. Results. Groups 1 (CH only) and 2 (CH?+?HD) had significantly (p?<?0.05) more HGF-positive nonparenchymal cells than the control group. Also, specimens from CH-only patients (Group 1) had significantly (p?<?0.05) more positive staining than those from CH patients on HD (Group 2). Groups 1 and 2 had similar results for grading of CH and fibrosis score. No association was found between the number of HGF-positive cells and necroinflammatory or fibrosis score. Conclusions. The expression of HGF in liver tissue in CH patients on HD was much lower than in CH-only patients. The extrahepatic tissue might be the source of increased HGF serum levels in HD patients.     

HGF/SF对姜黄素诱导大肠癌细胞凋亡的作用

目的 观察肝细胞生长因子／离散因子（HGF／SF）在姜黄素诱导的大肠癌细胞凋亡中的作用。方法 采用MTT法分析姜黄素对大肠癌细胞的抑制作用；流式细胞术评价HGF抗凋亡作用。结果不同浓度姜黄素作用Caco2细胞后发现，只在64μg／ml浓度发挥作用，即抑制细胞增殖；而同时加入不同浓度的HGF后可以拮抗姜黄素引起的Caco-2细胞生长的抑制作用，但HGF发挥作用无浓度依赖关系。流式细胞术检测结果发现，姜黄素只在64μg／ml浓度下诱导Caco-2细胞凋亡，在加入不同浓度HGF后，凋亡明显减少，但并无剂量依赖关系。MAPK途径在HGF拮抗姜黄素诱导的Caco-2细胞凋亡中的作用发现，阻断p42／p44MAPK和p38MAPK后，并不影响HGF对Caco-2细胞凋亡的保护作用。结论 HGF拈抗姜黄素诱导的Caco-2细胞凋亡，MAPK信号传导途径可能不参与此过程。     

低氧诱导因子1α及血管内皮生长因子在前列腺癌中的表达及意义

目的:观察低氧诱导因子1α(H IF-1α)及血管内皮生长因子(VEGF)在前列腺癌(PCa)中的表达及意义。方法:32例PCa患者,根据G leason评分,将≥7分者设为高G leason评分组(n=12),<7分者为低G leason评分组(n=20)。良性前列腺增生(BPH)16例,BPH伴高级别前列腺上皮内瘤(PIN)15例,正常前列腺组织(NP)12例。采用免疫组化染色CD34观察各组组织中微血管密度(MVD)及H IF-1α、VEGF的表达情况。结果:PCa、PIN中H IF-1α阳性表达率分别为62.5%、60.0%,较BPH(6.3%)及NP(0)高,差异有统计学意义(P<0.05)。PCa、PIN中VEGF阳性表达率分别为78.1%、73.3%,较BPH(18.7%)及NP(8.3%)高,差异亦有统计学意义(P<0.05)。PCa的MVD为66.9±18.0,明显高于BPH(28.3±6.9)及NP(15.3±2.9)(P<0.05)。高G leason评分组H IF-1α、VEGF阳性率及MVD值均高于低G leason评分组,差异均有显著性(P<0.05)。结论:H IF-1α及VEGF过表达是PCa形成的早期事件,与PCa密切相关。     

缺氧诱导因子-1与前列腺癌研究进展

缺氧诱导因子-1(HIF-1)与前列腺癌存在密切的关系。研究表明HIF-1在前列腺癌中不但与血管生成因子、增殖与存活因子、葡萄糖转运以及糖分解酶等有关,而且与p53、p21、信号转导通路等有关。通过对HIF-1的深入研究将对前列腺癌的诊断与治疗提供新的思路。现对HIF-1的结构、功能及与前列腺癌关系进行综述。     

Elevated Hepatocyte Growth Factor Levels at the Beginning of High-Flux Hemodialysis Are Due to Heparin Administration

Background. It has been reported that hemodialysis (HD) stimulates hepatocyte growth factor (HGF) release, but it is not clear if this stimulation is due to HD itself or to heparin used during HD. To clarify this issue, we undertook the present study. Methods. We studied 18 HD patients using high-flux dialyzers, during a single 4-hr hemodialysis session (session A). The dialyzers were pre-rinse with normal saline without heparin, and HD was started with zero ultrafiltration and without anticoagulation. Anticoagulation was administered as IV injection (80 IU/kg of LMWH enoxaparin sodium) 10 min after the beginning of HD. HD was continued for 10 more minutes and then as prescribed. HGF serum levels were measured before the beginning of the HD session (sample t0) as well as 10 and 20 minutes after the beginning of the session (samples t10 and t20). In six more patients (controls), the same study was repeated but without the administration of LMWH during the first 20 min of HD initiation (session B). Results. In comparison with t0, t10 HGF serum levels changed significantly in neither session A nor in session B. However, at t20, HGF levels increased significantly in session A compared with t0 (increment 666.3 ± 211.0%, p < 0.0001) and t10 (increment 894.2 ± 506.0%, p < 0.0001), but not in session B. No differences were found between sessions A and B at samples t0 and t10 (p?=?NS). HGF serum levels at t20 in session A were found to be higher compared with corresponding levels in session B (p < 0.0001). Conclusion. Elevated HGF serum levels at the beginning of high-flux HD session are due to LMWH administration.     

转化生长因子调控表皮生长因子受体在雄激素非依赖型前列腺癌细胞中表达的意义

目的 :探讨转化生长因子 (TGF)α对前列腺癌雄激素非依赖型细胞系中表皮生长因子受体 (EGFR)表达的调控。　方法 :采用逆转录 多聚酶链式反应和Western印迹法对TGFα刺激前列腺癌雄激素非依赖型细胞系PC3、ARCaP后EGFRmRNA表达及其蛋白水平进行定量分析。　结果 :TGFα引起PC3、ARCaP的EGFRmRNA表达升高 ,分别为 5 .0 1± 0 .4 5和 9.0 5± 0 .6 3,明显高于对照组 (P <0 .0 5 )。PC3细胞系TGFα治疗后EGFR蛋白水平为 2 .2 8±0 .5 3,明显高于对照组 (P <0 .0 5 ) ;而ARCaP细胞系TGFα治疗后EGFR仅为 1 .2 4± 0 .2 2 ,和对照组比较无差别 (P >0 .0 5 )。　结论 :TGFα可以明显提高前列腺癌细胞的EGFR表达量 ;TGFα EGFR自分泌环参与雄激素非依赖型前列腺癌的形成     

表皮生长因子受体,多胺与肺癌的关系

探讨表皮生长因子受体（ＥＧＦＲ）和多胺（ＰＡ）对人肺癌及正常肺组织生长、分化的影响。方法放射性配体结合法检测ＥＧＦＲ含量；高效液相色谱分析法测ＰＡ含量。结果肺癌组织中ＥＧＦＲ的含量（５．６２±４．２６ｆｍｏｌ／ｍｇ膜蛋白）高于非癌肺组织（３．９０５±２．２７９ｆｍｏｌ／ｍｇ膜蛋白），有显著性差异（Ｐ＜０．０１）；肺癌组织ＰＡ的含量亦高于正常肺组织（Ｐ＜０．０１）。结论ＥＧＦＲ和ＰＡ可促使肺癌发展，可作为肺癌的肿瘤标记物     

IV型胶原酶在不同转移潜能的前列腺癌细胞系中的表达及活性分析

目的:探讨前列腺癌细胞转移潜能与IV型胶原酶基质金属蛋白酶2(MMP-2)和基质金属蛋白酶9(MMP-9)的关系。方法:应用免疫组织化学SABC法,研究IV型胶原酶在不同转移潜能的前列腺癌细胞株LNCaP、C4-2B中表达变化。同时采用明胶酶谱法对这2种前列腺癌细胞系分泌的IV型胶原酶活性进行分析。结果:①免疫组织化学染色显示,LNCaP、C4-2B细胞中均有MMP-2、MMP-9表达,主要表达于细胞质中,C4-2B细胞表达明显强于LNCaP(P<0.01)。②人前列腺癌细胞LNCaP的条件培养液中仅能检测到极低的MMP-2和MMP-9活性,酶谱条带的积分A值分别为0.89±0.02和0.86±0.01,主要以酶原形式存在,而C4-2B细胞则表现较强的MMP-2、MMP-9活性,积分A值分别为96.32±4.36和33.89±1.84,其分泌的MMP-2则以活性形式为主,其分泌量显著高于LNCaP细胞(P<0.01)。结论:前列腺癌细胞中IV型胶原酶的表达和分泌,有助于癌细胞的侵袭和转移;前列腺癌细胞的侵袭转移潜能与其产生IV型胶原酶的能力密切相关。     

Hepatocyte growth factor and its receptor Met are induced in crescentic glomerulonephritis.

BACKGROUND: In experimental extracapillary glomerulonephritis (EG) podocytes migrate, proliferate and change phenotype, and play a pivotal role in crescent formation. Hepatocyte Growth Factor (HGF) is an injury-induced effector of tissue repair that causes cell migration, growth and transdifferentiation via its receptor Met. METHODS: In 11 patients with EG we measured serum levels of HGF and investigated whether serum induces the release of HGF by Peripheral Blood Mononuclear Cells (PBMC). In renal biopsies we studied the expression of Met. In cultured podocytes we studied Met expression, migration, growth and morphological changes induced by recombinant (r) HGF. RESULTS: In patients with EG average serum levels of HGF (0.73 ng/ml) were higher than in normal volunteers (N, 0.10 ng/ml, p<0.01) and in patients with non-crescentic glomerular disease (GD, 0.18 ng/ml, p<0.01). Serum of EG induced a significant HGF release by PBMC (mean 0.58 ng/ml) in comparison with serum of N and GD (0.07 and 0.06 ng/ml, respectively, both p<0.001). Met was strongly expressed in crescents. Cultured podocytes expressed Met, and rHGF induced in podocytes a time- and dose-dependent migration, growth and epithelial to mesenchymal transdifferentiation. CONCLUSIONS: These results suggest that HGF/Met system participates in the process of crescent formation by inducing podocyte migration, growth and mesenchymal transformation.     

Telomere Length and Telomerase Activity in Bladder and Prostate Cancer Cell Lines

Background: Specific repeats of oligonucleotides at the ends of chromosomes (telomeres) are shortened by cell division in somatic cells and are thought to be related to aging. Immortal cells such as germ-line cells or cancer cells have demonstrated increased activity of the telomere-elongating enzyme (telomerase). The length of the telomeres of these cells is stable regardless of cell division. We examined the telomere length and telomerase activity in 3 bladder (JTC30, JTC32, and T24) and 2 prostate cancer (LNCaP and DU145) cell lines.
Methods: Telomere lengths were evaluated by Southern blot analysis with a oligonucleotide probe, (TTAGGG)_S and telomerase activities were detected with a polymerase chain reaction-based assay method.
Results: Telomerase activity was detected in all of the cell lines. Each cell line had a specific telomere length. In 2 bladder cancer cell lines (JTC30 and JTC32), the telomere length decreased with increased passage of the cells.
Conclusion: The presence of telomerase may be a biological character of bladder and prostate cancers as well as other malignancies, although it does not always compensate telomere shortening.     

反义核酸对放疗诱导前列腺癌细胞VEGF高表达的抑制作用

目的 :探讨放疗过程中病人血清血管内皮生长因子 (VEGF)升高的机制 ,以及VEGF反义硫代寡核苷酸(AS ODN)对前列腺癌细胞VEGF表达分泌的抑制作用。　方法 :动态观察前列腺癌病人放疗期间血清VEGF变化 ;体外照射培养的前列腺癌PC3M细胞 ,观察放疗对肿瘤细胞VEGF表达分泌的诱导作用及AS ODN的抑制效应。　结果 :前列腺癌病人血清VEGF浓度在放疗开始后逐渐升高 ,15d左右达到峰值后开始下降 ,到 6 0d时基本恢复到正常。前列腺癌细胞受X线照射后VEGF的分泌较对照组明显升高 ,AS ODN加照射组癌细胞VEGF分泌量较单纯照射组显著下降 (P <0 .0 1)。　结论 :放疗可刺激前列腺癌细胞中VEGF的表达分泌 ,VEGFAS ODN对放疗诱导的VEGF高表达有较好的抑制作用     

Hepatocyte Growth Factor in Saliva of Patients with Renal Failure and Periodontal Disease

Background: Hepatocyte growth factor (HGF), endogenous cytokine with pleiotropic repairing and regeneration properties in relation to most tissues and organs, contributes to the progression of periodontal disease (PD). Furthermore, PD is a significant health problem in patients with chronic renal failure (CRF). The role of HGF in the development of PD in this specific population was not a subject of research so far. Material and methods: The following groups were enrolled in the study: (1) 26 chronic hemodialysis (HD) subjects, (2) 26 patients treated by continuous ambulatory peritoneal dialysis (CAPD), (3) 28 predialysis CRF patients, (4) 26 subjects with advanced PD (without coexisting diseases), and (5) 20 healthy subjects without PDs. HGF level in saliva was measured using the immunoenzymatic method. Gingival index, papillary bleeding index, plaque index, and the loss of clinical attachment level were evaluated. Results: The HGF level in saliva of HD patients was twice higher than in that of subjects with healthy periodontium. Direct relationships between proper HGF level in saliva and the indices GI, PBI, and PI in CAPD-treated patients and with more severe PD were shown. It was found that PD is most advanced in HD patients, moderately in CAPD-treated patients and to the smallest extent in predialysis CRF patients. Conclusions: The HGF level in mixed saliva is the index of PD progression in subjects without renal failure and in CAPD-treated patients. PD is common in renal failure patients and is a significant problem concerning general health status.