前列腺导管系统远近端的细胞凋亡、雄激素受体亚型与雄雌激素环境研究

目的研究前列腺导管系统远、近端组织的细胞凋亡率、雄激素受体(AR)亚型表达及其雄、雌激素环境,以探讨诸因素在前列腺增长和前列腺增生(BPH)发病中的作用。方法按照前列腺的解剖学特征,分别切取正常前列腺导管系统远、近端组织10例。应用DNA末端原位标记的方法,对正常前列腺导管系统的远近端组织以及20例BPH标本进行了细胞凋亡率的研究,应用聚丙烯酰胺凝胶等电聚焦电泳(IEF)技术分析了相应组织的AR亚型表达状态,并检测了相应组织的双氢睾酮(DHT)和雌二醇(E2)水平和表皮生长因子受体(EGFR)含量。结果 正常前列腺导管系统远近端组织的DHT和E2差异无显著性,BPH组织中的E2含量(平均6.61 ng/g蛋白)略高于正常前列腺导管系统近端组织的含量3.13 ng/g蛋白,差异无显著性(P>0.05),而雄激素受体亚型在正常前列腺导管系统远近端和BPH组织中有明显不同的表达特征,同时正常前列腺导管系统远端组织的细胞凋亡率(41.2±3.5)％显著高于导管系统近端组织(29.2±4.0)％(P<0.05),后者非常显著高于BPH组织(3.9±1.1)％(P<0.001)。正常与增生前列腺组织内的DHT及E2含量与细胞调亡率之间无明显相关性。细胞调亡率与其组织的DHT、E2无明显相关性。结论前列腺导管系统的远近端上皮细胞结构形态生物学特性不同,其周围的间质亦有     

雄激素影响前列腺组织bFGF表达的研究

雄激素影响前列腺组织ｂＦＧＦ表达的研究谢庆祥刘春晓姚华强汪鸿缪友仁应用免疫组化ＳＰ方法结合计算机辅助图像分析对雄激素影响鼠前列腺组织成纤维细胞生长因子（ｂＦＧＦ）的表达进行了研究。实验方法雄性ＳＤ鼠７０只，体重２５０±３０克，随机分成４组。Ａ组１０只...     

实验性糖尿病诱导青春期大鼠前列腺细胞凋亡及转化生长因子的表达与意义

目的探讨糖尿病对青春期大鼠前列腺组织发育的影响及其可能的机理。方法观察青春期糖尿病大鼠在成年后前列腺组织的形态结构的变化,并测定血清胰岛素及雄激素的含量。原位细胞凋亡标记检测前列腺细胞凋亡,免疫组化检测TGF-β1蛋白表达。结果糖尿病组的血睾酮和胰岛素明显下降(P<0.01)。前列腺腺体发生萎缩,细胞凋亡显著增加(P<0.01),且TGF-β1蛋白表达也明显升高(P<0.01)。结论糖尿病可导致体内激素水平发生变化而使前列腺发育受阻,并通过对TGF-β1的调控而诱导细胞凋亡。     

生长因子在大鼠前列腺组织中的表达及雄激素的调控作用

目的　观察雄激素水平改变对大鼠前列腺组织碱性成纤维细胞生长因子（ｂＦＧＦ） 和转化生长因子β１（ＴＧＦβ１）表达的影响。方法　采用分子杂交技术研究ｂＦＧＦｍＲＮＡ 及ＴＧＦβ１ｍＲＮＡ 在去势和未去势大鼠前列腺组织中表达的差异。结果　去势组ＴＧＦβ１ｍＲＮＡ 杂交斑点ＩＯＤ 均值为３１５．５,是未去势组（１７９）的１．７６ 倍;而未去势组ｂＦＧＦｍＲＮＡ 杂交斑点ＩＯＤ 均值为４４５ ．３,是去势组（２００．３） 的２．２２倍,差异均有显著性（Ｐ＜０ ．０１）。结论　雄激素水平变化可影响ｂＦＧＦｍＲＮＡ 及ＴＧＦβ１ｍＲＮＡ 的表达,ｂＦＧＦ及ＴＧＦβ１ 介导雄激素的作用。     

雄激素剥夺条件下阻断自噬增加LNCaP细胞凋亡

目的 探讨雄激素剥夺条件下阻断自噬后LNCaP细胞凋亡变化与半胱天冬酶(caspase)激活的关系.方法 应用激光共聚焦显微镜、RT-PCR方法观察雄激素剥夺致细胞自噬增加后,利用DAPI染色观察细胞凋亡变化及药物抑制caspase后对凋亡的影响.结果 ①雄激素去除后LNCaP细胞自噬体增加,标准培养基(CM)培养下LNCaP细胞自噬体数量为1.90分;无血清培养基(SF)中细胞自噬体数量增高为2.64分;加入双氧睾酮(SFA组)后细胞自噬体下降至1.85分(P<0.01).CM中LNCaP细胞LC3 mRNA表达率为23%,血清饥饿12 h后,LC3表达量上调至100%,而SFA组LC3 mRNA表达量为86%;血清饥饿24 h后,SF组LC3 mRNA表达量为62%,SFA组为35%.②SF组和SFA组LNCaP细胞基础凋亡率分别为(3.19±1.09)0A和(3.01±0.33)%,加入3-甲基腺嘌呤(3-MA)阻断自噬24 h后,SF组凋亡率为(10.90±2.91)%,SFA组为(4.63±1.69)%.SF+3-MA组中加入Z-VAD-FMK后,细胞凋亡减至(1.16±0.52)%.组间差异有统计学意义(P<0.01).结论 剥夺雄激素后LNCaP细胞中自噬明显增加,阻断自噬后凋亡发生率增加.     

雄激素剥夺条件下阻断自噬增加LNCaP细胞凋亡

Objective To investigate the change of apoptosis in LNCaP cells after inhibition of autophagic process under androgen removal conditions. Methods The autophagic level was deter-mined by using confocal microscopy and RT-PCR. The DAPI staining was used to indicate the apopto-sis of LNCaP cells after inhibition of autophagic by 3-MA. Also, Z-VAD-FMK was used to extend the apoptosis results. Results ①Androgen deprivation led to increased autophagy in LNCaP cells. LN-CaP cells cultured in complete medium(CM) presented low autophagic process with 1.9 scores. After 24 hours, the punetate GFP-LC3 structures were accumulated in the cells cultured in serum-free medi-um (SF)(2.64 scores). In contrast, the number of punctate GFP-LC3 remained at a very low level (1.85 scores), when cells were incubated with DHT in SFA(serum-free medium+DHT). Statistical analysis showed the significant difference between SF and SFA (P<0.01). Semiquantitative RT-PCR was employed to examine the mRNA expression of LC3. Indeed, cells grown in the medium without serum had a higher LC3 mRNA expression with the highest at 12 hour time point as compared with the cells grown in CM. DHT treatment reduced the level of LC3 mRNA. ②Blockage of autophagy by 3-MA increased the apoptosis of LNCaP cells. LNCaP cells in SF and SFA just presented a basal level of apoptosis, which is (3.19±1.09)% and (3.01±0.33)% , respectively. Under androgen-free con-ditions, inhibition of autophagy by 3-MA could increase apoptosis significantly(10. 90±2.91%). While Z-VAD-FMK, a pan Caspase inhibitor, was able to suppress this apoptotic process to the level of (1.16±0.52)%, which was statistically significant(P<0.01). Conclusions Androgen removal can lead to the increase of autophagy in LNCaP cells. Moreover, inhibition of autophagy promotes the occurrence of apoptosis.     

雄激素剥夺条件下阻断自噬增加LNCaP细胞凋亡

Objective To investigate the change of apoptosis in LNCaP cells after inhibition of autophagic process under androgen removal conditions. Methods The autophagic level was deter-mined by using confocal microscopy and RT-PCR. The DAPI staining was used to indicate the apopto-sis of LNCaP cells after inhibition of autophagic by 3-MA. Also, Z-VAD-FMK was used to extend the apoptosis results. Results ①Androgen deprivation led to increased autophagy in LNCaP cells. LN-CaP cells cultured in complete medium(CM) presented low autophagic process with 1.9 scores. After 24 hours, the punetate GFP-LC3 structures were accumulated in the cells cultured in serum-free medi-um (SF)(2.64 scores). In contrast, the number of punctate GFP-LC3 remained at a very low level (1.85 scores), when cells were incubated with DHT in SFA(serum-free medium+DHT). Statistical analysis showed the significant difference between SF and SFA (P<0.01). Semiquantitative RT-PCR was employed to examine the mRNA expression of LC3. Indeed, cells grown in the medium without serum had a higher LC3 mRNA expression with the highest at 12 hour time point as compared with the cells grown in CM. DHT treatment reduced the level of LC3 mRNA. ②Blockage of autophagy by 3-MA increased the apoptosis of LNCaP cells. LNCaP cells in SF and SFA just presented a basal level of apoptosis, which is (3.19±1.09)% and (3.01±0.33)% , respectively. Under androgen-free con-ditions, inhibition of autophagy by 3-MA could increase apoptosis significantly(10. 90±2.91%). While Z-VAD-FMK, a pan Caspase inhibitor, was able to suppress this apoptotic process to the level of (1.16±0.52)%, which was statistically significant(P<0.01). Conclusions Androgen removal can lead to the increase of autophagy in LNCaP cells. Moreover, inhibition of autophagy promotes the occurrence of apoptosis.     

雄激素剥夺条件下阻断自噬增加LNCaP细胞凋亡

Objective To investigate the change of apoptosis in LNCaP cells after inhibition of autophagic process under androgen removal conditions. Methods The autophagic level was deter-mined by using confocal microscopy and RT-PCR. The DAPI staining was used to indicate the apopto-sis of LNCaP cells after inhibition of autophagic by 3-MA. Also, Z-VAD-FMK was used to extend the apoptosis results. Results ①Androgen deprivation led to increased autophagy in LNCaP cells. LN-CaP cells cultured in complete medium(CM) presented low autophagic process with 1.9 scores. After 24 hours, the punetate GFP-LC3 structures were accumulated in the cells cultured in serum-free medi-um (SF)(2.64 scores). In contrast, the number of punctate GFP-LC3 remained at a very low level (1.85 scores), when cells were incubated with DHT in SFA(serum-free medium+DHT). Statistical analysis showed the significant difference between SF and SFA (P<0.01). Semiquantitative RT-PCR was employed to examine the mRNA expression of LC3. Indeed, cells grown in the medium without serum had a higher LC3 mRNA expression with the highest at 12 hour time point as compared with the cells grown in CM. DHT treatment reduced the level of LC3 mRNA. ②Blockage of autophagy by 3-MA increased the apoptosis of LNCaP cells. LNCaP cells in SF and SFA just presented a basal level of apoptosis, which is (3.19±1.09)% and (3.01±0.33)% , respectively. Under androgen-free con-ditions, inhibition of autophagy by 3-MA could increase apoptosis significantly(10. 90±2.91%). While Z-VAD-FMK, a pan Caspase inhibitor, was able to suppress this apoptotic process to the level of (1.16±0.52)%, which was statistically significant(P<0.01). Conclusions Androgen removal can lead to the increase of autophagy in LNCaP cells. Moreover, inhibition of autophagy promotes the occurrence of apoptosis.     

雄激素剥夺条件下阻断自噬增加LNCaP细胞凋亡

Objective To investigate the change of apoptosis in LNCaP cells after inhibition of autophagic process under androgen removal conditions. Methods The autophagic level was deter-mined by using confocal microscopy and RT-PCR. The DAPI staining was used to indicate the apopto-sis of LNCaP cells after inhibition of autophagic by 3-MA. Also, Z-VAD-FMK was used to extend the apoptosis results. Results ①Androgen deprivation led to increased autophagy in LNCaP cells. LN-CaP cells cultured in complete medium(CM) presented low autophagic process with 1.9 scores. After 24 hours, the punetate GFP-LC3 structures were accumulated in the cells cultured in serum-free medi-um (SF)(2.64 scores). In contrast, the number of punctate GFP-LC3 remained at a very low level (1.85 scores), when cells were incubated with DHT in SFA(serum-free medium+DHT). Statistical analysis showed the significant difference between SF and SFA (P<0.01). Semiquantitative RT-PCR was employed to examine the mRNA expression of LC3. Indeed, cells grown in the medium without serum had a higher LC3 mRNA expression with the highest at 12 hour time point as compared with the cells grown in CM. DHT treatment reduced the level of LC3 mRNA. ②Blockage of autophagy by 3-MA increased the apoptosis of LNCaP cells. LNCaP cells in SF and SFA just presented a basal level of apoptosis, which is (3.19±1.09)% and (3.01±0.33)% , respectively. Under androgen-free con-ditions, inhibition of autophagy by 3-MA could increase apoptosis significantly(10. 90±2.91%). While Z-VAD-FMK, a pan Caspase inhibitor, was able to suppress this apoptotic process to the level of (1.16±0.52)%, which was statistically significant(P<0.01). Conclusions Androgen removal can lead to the increase of autophagy in LNCaP cells. Moreover, inhibition of autophagy promotes the occurrence of apoptosis.     

Mechanisms of the development of androgen independence in prostate cancer

The effectiveness of androgen ablation in the management of advanced prostate cancer is of limited duration, with the median length of response being only 18–24 months. The transition of the prostate cancer cell to an androgen independent phenotype is a complex process that involves selection and outgrowth of pre-existing clones of androgen-independent cells (clonal selection) as well as adaptive up-regulation of genes that help the cancer cells survive and grow after androgen ablation (adaptation). These two mechanisms share an important pre-requisite characteristic: prostate cancers are heterogeneous tumours comprised of various subpopulations of cells that respond differently to androgen withdrawal therapy. This tumour heterogeneity may reflect either a multifocal origin, adaptation to environmental stimuli, and/or genetic instability of the initial cancer. This review will reexamine the different mechanisms that enable prostate cancer cells to proliferate in an androgen depleted environment.     

雄激素对前列腺癌细胞的双向作用及其机制

目的探讨雄激素对前列腺癌细胞及其双向作用分子机制。方法在LNCaP细胞培养液中加入10~(-9)、10~(-10)、10~(-11)、10~(-12) mol/L的人工合成雄激素R1881,使用噻唑蓝(MTT)比色法和流式细胞术证明其双相作用。分别提取受R1881刺激和抑制的LNCaP细胞中的mRNA,反转录后行基因芯片分析,并使用逆转录-聚合酶链反应(RT-PCR)技术验证其结果。结果细胞被10~(-9) mol/L及以上浓度的R1881抑制,被10~(-10)mol/L及以下浓度的R1881刺激生长。与对照组比较,10~(-9)mol/L R1881作用组,320基因表达差异有意义,170基因表达下调,150基因表达上调。10~(-10) mol/LR1881作用组中,4608条基因表达差异有意义,2562基因表达下调,2046基因上调。结论R1881可在10~(-9)mol/L及以上浓度抑制LNCaP细胞生长,在10~(-10)mol/L及以下浓度刺激细胞生长。雄激素受体共调节物在R1881的双向作用中可能起一定作用。     

Androgen and prostate cancer: the role of primary androgen deprivation therapy in localized prostate cancer

BackgroundThe basic mechanisms and clinical efficacy of primary androgen deprivation therapy (PADT), especially combined androgen blockade (CAB) for localized or locally advanced prostate cancer (PCa) have been outlined. An important point relates to which patients are suitable candidates for PADT.MethodsA retrospective review of the efficacy of PADT in 628 patients with localized or locally advanced PCa treated with PADT at seven institutions in Japan was carried out.ResultsIt was found that more than 30% of low- or intermediate-risk localized PCa patients could have their disease controlled over the long-term by PADT alone. Short-term or intermittent PADT could not be recommended because of the possibility of character change in the cancer cells as a result of incomplete androgen ablation.ConclusionAlgorithms are proposed for the treatment of localized PCa not only in low- and intermediate-risk groups, but also in the high-risk group. Future research directions are indicated.     

A soluble androgen receptor in the cytoplasm of the male mastomys prostate

Summary A steroid receptor protein was isolated from the cytoplasmic fraction of Mastomys prostate. Following in vivo and in vitro labelling of the tissue with tritiated testosterone or dihydrotestosterone, samples were analysed by gel exclusion chromatography or sucrose density gradient centrifugation. A steroid receptor complex was isolated on Sephadex G-200. Analysis of the steroids associated with this complex showed that the major part of the bound radioactivity was 5 -dihydrotestosterone. The binding was inhibited by unlabelled testosterone and could not be demonstrated in the liver cytosol. Using sucrose desity gradient centrifugation, the dihydrotestosterone receptor complex sedimented at 5.6 s together with heavier aggregates. In the presence of 0.4 M KCl a single complex was sedimented at 4. 6 s. The results demonstrate a receptor protein in the cytosol of the Mastomys prostate which binds to dihydrotestosterone and is comparable to that of rat prostate.     

雄激素受体基因ＣＡＧ多态性与前列腺癌的关系

目的：进一步探讨雄激素受体（ＡＲ)基因ＣＡＧ多态性与前列腺癌的关系,方法：采用分子生物学方法对３５８例前列腺癌患者ＡＲ基因ＣＡＧ进行测试,分析ＣＡＧ长度与临床各种指标的关系,结果：经统计学分析显示ＡＲ基因ＣＡＧ与年龄之间呈高相关性（Ｐ＝０．００７,r＝０．１１４）,但与ＰＳＡ、肿瘤分级及分期无显著相关性,结论：ＡＲ基因ＣＡＧ的长度与患者年龄成正比,即前列腺癌患者年龄越轻,其ＣＡＧ长度越短。     

Nuclear androgen receptors in the prostate of male Praomys (Mastomys) Natalensis

Summary The binding of dihydrotestosterone (DHT) within the nuclear fraction of the prostate of male Praomys (Mastomys) Natalensis has been investigated. Using in vivo or in vitro labelling with ³H-DHT, the presence of a receptor protein having a sedimentation coefficient of 3.0 S was demonstrated. The binding was shown to be specific towards DHT and could not be found in control tissue. Analysis of radiometabolites associated with the steroid receptor complex demonstrated that the majority of the bound steroid was DHT. The similarity between this steroid receptor complex and that of rat ventral prostate, together with the potential use of this experimental model are discussed.     

The role of estrogen receptor, androgen receptor and growth factors in diethylstilbestrol-induced programming of prostate differentiation

Recently, others and we have demonstrated that prenatal exposure to an extremely low dose of diethylstilbestrol (DES) and other estrogenic compounds produces a significant effect on mouse prostate development in vivo and in vitro in the presence and absence of androgen. In this study, we investigated the mechanism by which DES produces this effect and determined the role of its estrogenic activity on the growth and branching, induced by DES in the 17-day-old fetal prostate in culture. Additionally, we investigated whether the androgen receptor (AR) plays a role and whether any of the growth factors, namely, EGF and IGF-1 which are known to modulate the estrogen receptor (ER) and androgen receptor (AR)-dependent process, mediate the DES-induced effects. Using the organ culture bioassay of prostate development, we demonstrate that DES enhanced the growth and branching of the prostate at both 0.1 and 0.5 pg/ml dosages, thus, confirming a previous report of ours. An anti-estrogen, ICI164,387 blocked both of the effect of DES, suggesting that both of these two effects are ER dependent. Anti-androgen, flutamide also blocked both branching and prostatic growth induced by DES, while cyproterone acetate blocked only the branching effect, suggesting a role for AR in the DES-induced effects. Depletion of EGF by anti-EGF antibody blocked the DES-induced effects and this was reversed following EGF replacement in the organ culture system. Anti-IGF-1 antibody, on the other hand, only blocked the branching effect, but produced no effect on the prostatic growth, induced by DES. Estrogenic chemicals, bisphenol A and DES enhanced EGF-mRNA level of the cultured prostates. Taken together, it appears that DES-induced prostatic enlargement involves enhancement of ER-dependent EGF and IGF-1 synthesis, mediating prostatic enlargement and androgen action. Received: 21 July 1999 / Accepted: 1 February 2000     

三维适形放疗加内分泌联合治疗晚期前列腺癌

目的评价三维适形放疗加内分泌联合治疗晚期前列腺癌的效果。方法对晚期前列腺癌患者25例行3DCRT 内分泌联合治疗(联合组),以同期进行的单纯内分泌治疗的晚期前列腺癌患者40例为对照组。随访时间3~48个月,中位随访期27个月。结果联合组3年生存率为88.0%,明显高于对照组(68.0%)。在30个月后,联合组的PSA低于对照组,差异有统计学意义(P<0.05)。结论3DCRT 内分泌联合治疗晚期前列腺癌疗效满意,优于单纯内分泌治疗。     

Gender differences in expression of androgen receptor in tibial growth plate and metaphyseal bone of the rat

In this study, we investigate the expression of the androgen receptor (AR) in the tibial growth plate and metaphyseal bone of male and female rats at the mRNA and protein level. Using in situ hybridization and immunohistochemistry, AR mRNA and protein were demonstrated in proliferating and early hypertrophic chondrocytes in the growth plate of 1-, 4-, and 7-week-old male and female rats. Immunostaining for AR was observed both in the nucleus and the cytoplasm. After sexual maturation at 12 and 16 weeks of age, AR expression decreased in both genders and was confined to a small rim of prehypertrophic chondrocytes. In female rats of 40 weeks of age, this expression pattern was still visible. In most age groups there was a tendency toward an increased AR mRNA expression in male vs. female rats except in the 7-week-old animals. At the protein level, sexually maturing 7-week-old male rats demonstrated a higher staining intensity compared to their female counterparts. At this stage, AR staining in the males was mainly confined to the nucleus, whereas in females staining was predominantly found in the cytoplasm. In the tibial metaphysis, AR mRNA was detected in lining cells, osteoblasts, osteocytes, and osteoclasts at all stages of development. At the protein level, a similar expression pattern was observed, except for an absence of immunostaining in the lining cells. The staining was both nuclear and cytoplasmic. In most age groups, mRNA and protein signals were higher in males compared with females. We have demonstrated the presence of AR mRNA and protein in the tibial growth plate and the underlying metaphyseal bone during development of the rat. In male rats, the presence of higher messenger and protein staining intensities, as well as preferential nuclear staining during sexual maturation, suggests that direct actions of androgens in chondrocytes and in bone forming cells may be involved in establishing the gender differences in the skeleton.     

雄激素共激活物DDC在氟他胺对前列腺癌细胞LNCaP作用中的表达变化

目的 观察雄激素受体（AR）共激活物DDC在氟他胺对前列腺癌雄激素敏感细胞系LNCaP的作用的表达变化。方法 噻唑蓝比色法（MTT），流式细胞术等技术，研究氟他胺对LNCaP细胞的作用，并使用基因芯片对其DDC在其中的作用作了初步探讨。结果 LNCaP细胞在7d内受到氟他胺的刺激而加速生长，但5代后细胞增殖受到抑制。此种效应与剂量相关。细胞增殖主要被阻断于G1／S期。其相关机制主要是AR共激活物DDC的表达受到抑制。结论 氟他胺对LNCaP的作用为双向，即在低浓度和较短作用时间内为增殖，在高浓度和较长作用时间后为抑制。此种抑制效应可能是通过抑制DDC的表达，影响下游细胞周期相关基因的表达，从而阻断细胞周期于G1／S调控点。